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Sample records for igm antibody determination

  1. Determination of Antibodies (IgG, IgM against Toxoplasma gondii in Patients with Cancer

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    M Pedram

    2007-08-01

    Full Text Available Background: The aim of this study was determination of antibodies (IgG, IgM against Toxoplasma in malignant patients in order to refer the patients on time to the physician for treatment.Methods: This study was carried out on 252 malignant patients and 252 healthy normal subjects (as control obtained from Shafa Hospital and Medical Diagnostic Laboratory (Iran-Zamin, in Ahwaz city. Patient's information was recorded in a questionnaire before sampling. Serum samples of patients were examined for IgG and IgM antibodies by ELISA technique using Trinity kits. Results: The results of this study revealed the presence of Toxoplasma antibodies in 114 (45.2% cases of patients who were positive for Toxoplasma IgG antibodies, and 26 (10.3% cases were confirmed to be positive for Toxoplasma IgM antibodies and also 17 (6.7% of cases had both IgG and IgM antibodies against Toxoplasma gondii. In control group 92 (36.5% cases and 15 (6% cases revealed seropositive for IgG and IgM antibodies, respectively. There were no significant differences between sex, close contact with cat, living region, chemotherapy, and seropositivity rate of toxoplasmosis in patients. Comparing the age groups, the highest seropositive rate showed in the age of 51 years or higher, and their rates had tendency to increase with age in both groups. No seropositivity significant relationship was found between patients and control group.Conclusion: According to the prevalence of positive cases in these patients, it is necessary to examine the patients for toxoplasmosis before, during and after chemotherapy.

  2. Determination of rubella IgG and IgM antibodies among women of ...

    African Journals Online (AJOL)

    Rubella is an important Toga virus disease, causing significant human public health problem in Nigeria. In this study effort was made to determine rubella prevalence in women in Kano State. One thousand blood sera were tested with IgM specific Enzyme Immuno Assay (EIA) and IgG indirect solid phase ELISA based kits.

  3. IgM antibodies against dsDNA in SLE.

    Science.gov (United States)

    Witte, Torsten

    2008-06-01

    IgG antibodies against dsDNA are involved in the pathogenesis of systemic lupus erythematosus (SLE) glomerulonephritis. In contrast, glomerulonephritis is rare in SLE patients with IgM antibodies against dsDNA. Therefore, a possible protective effect of IgM antibodies has been studied in more detail. In murine models of SLE, the lack of secreted IgM was associated with more severe glomerulonephritis. In more recent studies, the treatment of lupus-prone mice with a murine IgM monoclonal antibody against dsDNA prevented renal damage. Furthermore, the clearance of pathogenic immune complexes may be improved by IgM. Therefore, IgM antibodies against dsDNA are indeed protective and may be a new treatment modality of lupus nephritis in humans.

  4. Antibodies recognizing both IgM isotypes in Atlantic salmon

    DEFF Research Database (Denmark)

    Hedfors, Ida Aagård; Bakke, Hege; Skjødt, Karsten

    2012-01-01

    these molecules. The present study aimed at identifying tools to separate IgM positive (IgM(+)) B cells from IgM negative (IgM(-)) non-B cell populations using flow cytometry. Several monoclonal antibodies (mAbs), and one polyclonal antibody (pAb) to both rainbow trout (Oncorhynchus mykiss) and Atlantic salmon...... (Salmo salar) IgM, either commercially available or locally produced were tested for their recognition of Atlantic salmon IgM(+) cells. Leukocytes were isolated from peripheral blood (PB), spleen (S) and head kidney (HK) and stained with all mAbs and the pAb, to possibly verify the approximate number...... of IgM(+) cells in the respective tissues in salmon. To our surprise, this seemingly simple task did not reveal similar staining patterns for all antibodies as expected, but rather large differences in the number of positively stained cells were discovered. In short, positively stained cells by each...

  5. [IgM antiphospholipical antibodies in preeclampsia-eclampsia].

    Science.gov (United States)

    Briones-Garduño, Jesús Carlos; Díaz de León-Ponce, Manuel; Barrios-Prieto, Ernesto; Salazar-Exaire, José Daniel

    2003-01-01

    Antiphospholipid antibodies are associated with thrombocytopenia and fetal loss, and have been reported elevated in patients with preeclampsia-eclampsia. Site: Preeclampsia-Eclampsia Research Unit, Instituto Materno Infantil del Estado de Mexico, Toluca, Mexico and the Specialty Hospital Research Unit, La Raza Medical Center, Mexico City, Mexico. To determine the presence of anticardiolipin antibodies (IgG-IgM) as markers of acute endothelial damage in patients with preeclampsia. A randomized case control study composed of two groups: Group A (cases), 18 patients with preeclampsia-eclampsia and group B (control), 18 normal pregnancies. Antiphospholipid (anticardiolipin) antibodies were determined in both groups in addition to with coagulation tests and clinical variables in mother and newborn in day of admission and nine weeks after obstetrical resolution. We excluded patients with anticoagulant or dialysis therapy, taking NSAIDs or who recently required transfusion or plasmaferesis. There were significant differences in levels of IgM and in weights and mortality among newborns between the two groups. In terms of maternal complications, we found HELLP syndrome as leading cause. We also observed in the case group significant differences in mean arterial pressure (MAP) and IgM levels between admission day and nine weeks later. Our results lead us to the conclusion that there must be exist immunologic mechanism that induces synthesis of anticardiolipin antibodies (IgM isotype) during acute state of the disease, accounting for vascular changes and prothrombotic state responsible for maternal and neonatal complications.

  6. The IgM Response to Modified LDL in Experimental Atherosclerosis Hypochlorite-modified LDL IgM Antibodies versus Classical Natural T15 IgM Antibodies

    NARCIS (Netherlands)

    van Leeuwen, Marcella; Damoiseaux, Jan; Duijvestijn, Adriaan; Heeringa, Peter; Gijbels, Marion; de Winther, Menno; Tervaert, Jan Willem Cohen; Shoenfeld, Y; Gershwin, ME

    2009-01-01

    Introduction: It is hypothesized that IgM antibodies to oxidized LDL are anti-atherogenic. Myeloperoxidase from plaque-infiltrating neutrophils catalyzes the production of hypochlorite (HOCl), which oxidizes LDL. Here we study the IgM response to HOCl-modified LDL in comparison to titers of T15

  7. Protective roles of natural IgM antibodies

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    Caroline eGrönwall

    2012-04-01

    Full Text Available Antibodies are a vital part of the armentarium of the adaptive immune system for the fine-tuning of the recognition and response to foreign threats. However, in health there are some types of antibodies that instead recognize self-antigens for the enhancement of primitive innate functions. The repertoire of natural IgM antibodies is postulated to have been selected during immune evolution for their contributions to critical immunoregulatory and housekeeping properties. The clearance of dying cells is one of the most essential responsibilities of the immune system, which is essential to prevent uncontrolled inflammation and autoimmunity. In the murine immune system, natural IgM antibodies that recognize apoptotic cells have been shown to enhance the phagocytic clearance of dead and dying cells and to suppress innate immune signaling pathways. In the mouse, natural IgM are often the products of B-1 cell clones that arise during immune development without an absolute requirement for exogenous antigenic stimulation. In patients with systemic lupus erythemtosus, IgM autoantibodies, which bind to neo-epitopes on apoptotic cells, have been demonstrated to be present at significantly higher levels in patients with lower disease activity and with less severe organ damage. While certain specificities of IgM autoantibodies correlate with protection from lupus renal disease, others may convey protective properties from lupus-associated atherosclerotic cardiovascular disease. New unexpected insights into the functional roles of IgM antibodies are still emerging, especially regarding the functions of natural antibodies. Herein, we review recent progress in our understanding of the potential roles of natural IgM autoantibodies in the regulation of immune homeostasis and for protection from autoimmune and inflammatory diseases.

  8. Microangiopathic antiphospholipid antibody syndrome due to anti-phosphatidylserine/prothrombin complex IgM antibody.

    Science.gov (United States)

    Senda, Yumi; Ohta, Kazuhide; Yokoyama, Tadafumi; Shimizu, Masaki; Furuichi, Kengo; Wada, Takashi; Yachie, Akihiro

    2017-03-01

    Herein we describe a case of microangiopathic antiphospholipid syndrome (MAPS) due to anti-phosphatidylserine/prothrombin complex (aPS/PT) IgM antibody successfully treated with rituximab. A significant correlation was observed between the clinical course and the aPS/PT IgM antibody titer, which can rise earlier before the appearance of clinical symptoms. Rituximab can be safely and effectively used for MAPS. Although detection of only aPS/PT IgM antibody is rare, aPS/PT IgM antibody might be associated with the pathogenesis of MAPS and might be a useful marker of disease activity. © 2017 Japan Pediatric Society.

  9. Sero-prevalence of IgM antibody to Rubella Virus in pregnant ...

    African Journals Online (AJOL)

    Rubella is a contagious viral infection, which in pregnant women leads to infection of a developing fetus causing fetal death or Congenital Rubella Syndrome. A cross-sectional study involving 180 women was carried out between June and August 2012 to determine the seroprevalence of IgM antibody to rubella in their ...

  10. The detection of rubella-specific IgM antibodies by radioimmunoassay

    International Nuclear Information System (INIS)

    Kangro, H.O.; Pattison, J.R.; Heath, R.B.

    1978-01-01

    An indirect solid-phase radioimmunoassay (RIA) has been developed for the detection of immunoglobulin (Ig) class-specific rubella antibodies. A commercial rubella haemagglutinin is dried and fixed on to the wells of flexible microtitre plates and allowed to react with serial dilutions of whole or fractionated human sera. Class-specific rubella antibodies are then detected by determining the specific binding of 125 I-labelled anti-human IgG or IgM. The RIA was first evaluated by comparison with the heamagglutination-inhibition (HI) test for the detection of rubella-specific IgM in gel-filtration fractions. RIA was found to be as specific as HI but 50-150 times more sensitive. Rubella-specific IgG antibodies did not interfere in specific IgM determinations by RIA and therefore the latter technique applied to unfractionated sera. The results obtained indicate that RIA on unfractionated sera is a practical, sensitive and specific technique which could provide a reliable method for the diagnosis of rubella. The rubella-specific IgM titres obtained by RIA were not increased by the removal of IgG by pretreatment of sera with Staphylococcal Protein A. (author)

  11. Determination of total (IgG/IgM and specific (IgM antibodies to Hepatitis E Virus and molecular detection of the virus in feces of humans with or without occupational exposure to pigs in 10 municipalities of Antioquia, Colombia

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    Gutiérrez-Vergara, Cristian Camilo

    2015-07-01

    Full Text Available In 10 municipalities of Antioquia (Colombia the positivity rate in serum for total (IgG/IgM and specific (IgM antibodies to hepatitis E virus (HEV was determined, and tests were done for the presence of HEV RNA in the feces of individuals positive for IgM antibodies. According to previous exposure to pigs, two different groups were included, namely: exposed and unexposed. The latter group was subdivided into cohabitants of the exposed ones and general population. The frequency of total anti-HEV antibodies in the exposed group was 15.7%, and that of IgM, 2.5% (p<0.001. In the group of cohabitants, total antibodies were found in 5.9%, while IgM antibodies were not present. In the general population IgG/IgM antibodies were present in 7.2% and IgM, in 0.81% (p<0.001. None of the fecal specimens was positive for HEV RNA. These results indicate that individuals with occupational exposure to pigs have higher risk (RP: 2.42 of being positive for anti-HEV antibodies than the unexposed ones (95% CI: 1.66-3.53 (p<0.001. Also, that in Antioquia HEV is present regardless of the exposure to pigs. Further studies on HEV in Colombia should be done.

  12. Capture ELISA for IgM antibodies against Plasmodium falciparum glutamate rich protein

    DEFF Research Database (Denmark)

    Dziegiel, Morten Hanefeld; Borre, M B; Petersen, E

    1992-01-01

    This report describes a novel mu chain capture ELISA for the detection of IgM antibodies against a Plasmodium falciparum antigen. A fragment of the 220 kDa P. falciparum glutamate rich protein containing amino acid residues 489-1271 was expressed in E. coli as a recombinant chimeric beta......-galactosidase fusion protein and used as antigen after purification and biotinylation. Specific IgM antibodies were found in 51% (39/77) of sera from adult Liberians immune to malaria. The binding of IgM antibodies was specific for the malaria portion of the fusion protein and no cross-reactivity was found in sera...... from patients with IgM antibodies due to other diseases. Inhibition studies with a fusion protein containing amino acid residues 816-1134 (GLURP816-1134) representing the carboxy-terminal repeat region suggested a different use of epitopes for IgM antibodies in different individuals....

  13. Detection of measles IgM antibodies in children at Kaduna ...

    African Journals Online (AJOL)

    Measles virus IgM antibodies virus was assayed for in 270 sera from children aged 5 months to 8 years attending Yusuf Dantsoho Memorial Hospital, Kaduna, Northern Nigeria, using ELISA. Out of the 270 sera, 192(71.1%) tested positive to measles IgM and 78(28.9%) negative. The sample distribution was 137 from males, ...

  14. Breast cancer imaging with In-111 human IgM monoclonal antibodies: preliminary studies.

    Science.gov (United States)

    Ryan, K P; Dillman, R O; DeNardo, S J; DeNardo, G L; Beauregard, J; Hagan, P L; Amox, D G; Clutter, M L; Burnett, K G; Rulot, C M

    1988-04-01

    Detection of specific tumor sites was studied with scintigraphy and radiolabeled human IgM monoclonal antibodies (MoAbs). Ten patients with metastatic breast cancer received an infusion of one of three indium-111-labeled anti-breast carcinoma MoAbs. The time of infusion ranged from 30 minutes to 2 hours. Three patients received YBB-190 at total doses of 2, 4.25, or 11 mg, four patients received YBM-209 at total doses of 1 mg (n = 1) or 20 mg (n = 3), and three patients each received 22 mg of YBY-088. Imaging was performed immediately after infusion and at 4, 24, 48, 72, 120, and 144 hours. Many presumed sites of metastatic disease were imaged in three of the four patients who received 20 mg of YBM-209 and in two of the three patients who received YBY-088. Tumor was not detected in any of the patients who received YBB-190, in the patient who received a 1-mg dose of YBM-209, or in the patient who received YBY-088 and in whom a biopsy of tumor tissue failed to demonstrate target antigen. The authors conclude that In-111-labeled human IgM MoAbs can target human breast cancer, but antigen expression and antibody dose determine successful immunoscintigraphy.

  15. Human Secretory IgM Antibodies Activate Human Complement and Offer Protection at Mucosal Surface.

    Science.gov (United States)

    Michaelsen, T E; Emilsen, S; Sandin, R H; Granerud, B K; Bratlie, D; Ihle, O; Sandlie, I

    2017-01-01

    IgM molecules circulate in serum as large polymers, mainly pentamers, which can be transported by the poly-Ig receptor (pIgR) across epithelial cells to mucosal surfaces and released as secretory IgM (SIgM). The mucosal SIgM molecules have non-covalently attached secretory component (SC), which is the extracellular part of pIgR which is cleaved from the epithelial cell membrane. Serum IgM antibodies do not contain SC and have previously been shown to make a conformational change from 'a star' to a 'staple' conformation upon reaction with antigens on a cell surface, enabling them to activate complement. However, it is not clear whether SIgM similarly can induce complement activation. To clarify this issue, we constructed recombinant chimeric (mouse/human) IgM antibodies against hapten 5-iodo-4-hydroxy-3-nitro-phenacetyl (NIP) and in addition studied polyclonal IgM formed after immunization with a meningococcal group B vaccine. The monoclonal and polyclonal IgM molecules were purified by affinity chromatography on a column containing human SC in order to isolate joining-chain (J-chain) containing IgM, followed by addition of excess amounts of soluble SC to create SIgM (IgM J+ SC+). These SIgM preparations were tested for complement activation ability and shown to be nearly as active as the parental IgM J+ molecules. Thus, SIgM may offer protection against pathogens at mucosal surface by complement-mediated cell lysis or by phagocytosis mediated by complement receptors present on effector cells on mucosa. © 2016 The Foundation for the Scandinavian Journal of Immunology.

  16. Circulating microparticles carry oxidation-specific epitopes and are recognized by natural IgM antibodies.

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    Tsiantoulas, Dimitrios; Perkmann, Thomas; Afonyushkin, Taras; Mangold, Andreas; Prohaska, Thomas A; Papac-Milicevic, Nikolina; Millischer, Vincent; Bartel, Caroline; Hörkkö, Sohvi; Boulanger, Chantal M; Tsimikas, Sotirios; Fischer, Michael B; Witztum, Joseph L; Lang, Irene M; Binder, Christoph J

    2015-02-01

    Oxidation-specific epitopes (OSEs) present on apoptotic cells and oxidized low density lipoprotein (OxLDL) represent danger-associated molecular patterns that are recognized by different arcs of innate immunity, including natural IgM antibodies. Here, we investigated whether circulating microparticles (MPs), which are small membrane vesicles released by apoptotic or activated cells, are physiological carriers of OSEs. OSEs on circulating MPs isolated from healthy donors and patients with ST-segment elevation myocardial infarction (STE-MI) were characterized by flow cytometry using a panel of OSE-specific monoclonal antibodies. We found that a subset of MPs carry OSEs on their surface, predominantly malondialdehyde (MDA) epitopes. Consistent with this, a majority of IgM antibodies bound on the surface of circulating MPs were found to have specificity for MDA-modified LDL. Moreover, we show that MPs can stimulate THP-1 (human acute monocytic leukemia cell line) and human primary monocytes to produce interleukin 8, which can be inhibited by a monoclonal IgM with specificity for MDA epitopes. Finally, we show that MDA(+) MPs are elevated at the culprit lesion site of patients with STE-MI. Our results identify a subset of OSE(+) MPs that are bound by OxLDL-specific IgM. These findings demonstrate a novel mechanism by which anti-OxLDL IgM antibodies could mediate protective functions in CVD. Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

  17. Capture ELISA for IgM antibodies against Plasmodium falciparum glutamate rich protein

    DEFF Research Database (Denmark)

    Dziegiel, Morten Hanefeld; Borre, M B; Petersen, E

    1992-01-01

    This report describes a novel mu chain capture ELISA for the detection of IgM antibodies against a Plasmodium falciparum antigen. A fragment of the 220 kDa P. falciparum glutamate rich protein containing amino acid residues 489-1271 was expressed in E. coli as a recombinant chimeric beta-galactos......This report describes a novel mu chain capture ELISA for the detection of IgM antibodies against a Plasmodium falciparum antigen. A fragment of the 220 kDa P. falciparum glutamate rich protein containing amino acid residues 489-1271 was expressed in E. coli as a recombinant chimeric beta...

  18. Radioimmunoassay of IgM, IgG, and IgA brucella antibodies

    International Nuclear Information System (INIS)

    Parrett, D.; Nielson, K.H.; White, R.G.; Payne, D.J.H.

    1977-01-01

    A radioimmunoassay (R.I.A.) has been devised to measure the serum antibody against Brucella abortus in each of the immunoglobulin classes IgM, IgG, and IgA. This test was applied to 46 sera from individuals with various clinical types of brucellosis, and the results were compared with the results of conventional direct and indirect agglutination and complement-fixation tests. The R.I.A. provided a highly sensitive primary-type assay which avoided the difficulties with blocking or non-agglutinating antibody, and thus has many advantages in the diagnosis of acute and chronic stages of brucella infection in man. The R.I.A. was successful in detection of antibody in many instances in which conventional serological tests were negative, and such antibody could (if IgM) be associated with acute or (if IgG or IgA) with chronic cases of brucellosis. One case in which B.abortus was isolated by blood culture but which failed to yield antibody by conventional tests, nevertheless showed substantial levels of IgM and IgG antibody by R.I.A. In other cases the R.I.A. test helped to eliminate the diagnosis of brucellosis by revealing absent or low antibody levels. (author)

  19. Sero-prevalence of IgM antibody to Rubella Virus in pregnant ...

    African Journals Online (AJOL)

    Arun Kumar Agnihotri

    2014-09-05

    Sep 5, 2014 ... Sero-‐prevalence of IgM antibody to. Rubella Virus in pregnant women in. Zaria, Nigeria. Okikiola Olajide1, Maryam Aminu1, Abdullahi J Randawa2,. Daniel S Adejo2. ABSTRACT. Rubella is a contagious viral infection, which in pregnant women leads to infection of a developing fetus causing fetal death or ...

  20. Levels of natural IgM antibodies against phosphorylcholine in healthy individuals and in patients undergoing isolated limb perfusion

    NARCIS (Netherlands)

    Padilla, Niubel Diaz; Ciurana, Caroline; van Oers, Joep; Ogilvie, Aernout C.; Hack, C. Erik

    2004-01-01

    Natural IgM antibodies against phosphorylcholine (anti-Pc IgM) resemble C-reactive protein (CRP) regarding specificity and have gained increasing attention because of their supposed role in clearance of damaged cells and in cardiovascular disease. In order to quantify these antibodies in human

  1. Radioimmunoassay of IgM and IgG antitetanus toxoid antibody

    International Nuclear Information System (INIS)

    Repetti, C.F.; Gill, T.J. III

    1980-01-01

    A radioimmunoassay was developed to quantitate antitetanus toxoid antibody in whole plasma which can detect 0.75 ng IgG antibody (0.75 μg/ml plasma) and 1.60 ng IgM antibody (0.40 μg/ml plasma). The IgG antibody was measured by direct binding to a tetanus toxoid-Sepharose immunoadsorbent. The IgM antibody was measured by inhibition of binding by soluble tetanus toxoid because of the relatively high and erratic non-specific reaction between IgM and the immunoadsorbent. The immunoglobulin standards were [ 131 I]IgG or [ 131 I]IgM coupled to Sepharose, respectively, and the amount of antibody activity in the 125 I-labeled antiglobulin reagents could be calculated from the amount bound to the immunoadsorbent and the specific activity of the appropriate immunoglobulin standard. The assay is sensitive, reproducible and suitable for use with large numbers of samples: it should find wide applicability in both clinical and experimental settings. (Auth.)

  2. The Induction of IgM and IgG Antibodies against HLA or MICA after Lung Transplantation

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    Annelieke W. M. Paantjens

    2011-01-01

    Full Text Available The production of IgG HLA antibodies after lung transplantation (LTx is considered to be a major risk factor for the development of chronic rejection, represented by the bronchiolitis obliterans syndrome (BOS. It has recently been observed that elevated levels of IgM HLA antibodies also correlates with the development of chronic rejection in heart and kidney transplantation. This study investigates the relationship between IgM and IgG antibodies against HLA and MICA after lung transplantation. Serum was collected from 49 patients once prior to transplantation and monthly for up to 1 year after lung transplantation was analyzed by Luminex to detect IgM and IgG antibodies against HLA and MICA. The presence of either IgM or IgG HLA and/or MICA antibodies prior to or after transplantation was not related to survival, gender, primary disease, or the development of BOS. Additionally, the production of IgG alloantibodies was not preceded by an increase in levels of IgM, and IgM levels were not followed by an increase in IgG. Under current immune suppressive regimen, although the presence of IgM antibodies does not correlate with BOS after LTx, IgM high IgG low HLA class I antibody titers were observed more in patients with BOS compared to patients without BOS.

  3. Generation and characterization of antibodies against Asian elephant (Elephas maximus IgG, IgM, and IgA.

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    Alan F Humphreys

    Full Text Available Asian elephant (Elephas maximus immunity is poorly characterized and understood. This gap in knowledge is particularly concerning as Asian elephants are an endangered species threatened by a newly discovered herpesvirus known as elephant endotheliotropic herpesvirus (EEHV, which is the leading cause of death for captive Asian elephants born after 1980 in North America. While reliable diagnostic assays have been developed to detect EEHV DNA, serological assays to evaluate elephant anti-EEHV antibody responses are lacking and will be needed for surveillance and epidemiological studies and also for evaluating potential treatments or vaccines against lethal EEHV infection. Previous studies have shown that Asian elephants produce IgG in serum, but they failed to detect IgM and IgA, further hampering development of informative serological assays for this species. To begin to address this issue, we determined the constant region genomic sequence of Asian elephant IgM and obtained some limited protein sequence information for putative serum IgA. The information was used to generate or identify specific commercial antisera reactive against IgM and IgA isotypes. In addition, we generated a monoclonal antibody against Asian elephant IgG. These three reagents were used to demonstrate that all three immunoglobulin isotypes are found in Asian elephant serum and milk and to detect antibody responses following tetanus toxoid booster vaccination or antibodies against a putative EEHV structural protein. The results indicate that these new reagents will be useful for developing sensitive and specific assays to detect and characterize elephant antibody responses for any pathogen or vaccine, including EEHV.

  4. [Clinical aspects of children presenting specific IgM antibodies to cytomegalovirus by immunofluorescent test].

    Science.gov (United States)

    Yamamoto, A Y; Gonçalves, A L; Figueiredo, L T; Carlucci, R H

    1994-01-01

    Clinical data observed in 25 children presenting IgM specific antibodies to cytomegalovirus (CMV) by immunofluorescent test are reported. The children were grouped by the presented clinical picture in: neonatal hepatitis, mononucleosis syndrome,pneumonitis, neurologic disease, ocular abnormalities, and thrombocytopenic purpure. Clinical aspects of CMV infection were analyzed and compared with data reported in the literature. CMV infections can involve multiple organs and this viral disease must to be considered on differential diagnosis of many infectious diseases.

  5. Production and characterization of monoclonal antibodies to IgM of Pacific herring (Clupea pallasii)

    Science.gov (United States)

    Purcell, Maureen K.; Bromage, Erin S.; Silva, Jessica; Hansen, John D.; Badil, Samantha M.; Woodson, James C.; Hershberger, Paul K.

    2012-01-01

    Pacific herring (Clupea pallasii) have a central role in the North Pacific ecosystem as a forage fish species and are natural reservoirs of several important finfish pathogens, including Viral hemorrhagic septicemia virus (VHSV). Here, we report the identification of the gene encoding the immunoglobulin mu (IgM) heavy chain, as well as the development and characterization of monoclonal antibodies (MAbs) that specifically react with Pacific herring IgM. Pacific herring immunoglobulin was purified and consisted of heavy and light chains of approximately 80 and 25 kDa. Three hybridoma clones were initially identified by ELISA as reactive with purified immunoglobulin but only one clone was able to detect an 80 kDa protein in Pacific and Atlantic herring (Clupea harengus) whole plasma by denaturing western blot. However, all three MAbs were able to precipitate an 80 kDa protein from Pacific herring and LCMS sequencing of peptide fragments derived from this protein matched the predicted amino acid sequence of the cloned, heavy chain gene. In addition, two of the MAbs stained cells within the putative lymphocyte gates for the spleen, anterior kidney and posterior kidney but were not reactive for myeloid/granulocyte gates, which is consistent with these MAbs reacting with surface IgM+ B-cells. To our knowledge, this is the first report of IgM-related gene sequences and anti-IgM monoclonal antibodies from any member of the family Clupeidae. The antibodies produced in this study are critical for achieving our long-term goal of conducting serological surveillance to assess pathogen exposure in natural populations of Pacific herring.

  6. Destructive arthritis in a patient with chikungunya virus infection with persistent specific IgM antibodies

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    Receveur Marie-Catherine

    2009-12-01

    Full Text Available Abstract Background Chikungunya fever is an emerging arboviral disease characterized by an algo-eruptive syndrome, inflammatory polyarthralgias, or tenosynovitis that can last for months to years. Up to now, the pathophysiology of the chronic stage is poorly understood. Case presentation We report the first case of CHIKV infection with chronic associated rheumatism in a patient who developed progressive erosive arthritis with expression of inflammatory mediators and persistence of specific IgM antibodies over 24 months following infection. Conclusions Understanding the specific features of chikungunya virus as well as how the virus interacts with its host are essential for the prevention, treatment or cure of chikungunya disease.

  7. Genus and species-specific IgG and IgM antibodies pulmonary tuberculosis

    International Nuclear Information System (INIS)

    Butt, T.; Abbassi, S.A.; Ahmad, R.N.; Mahmood, A.; Karamat, K.A; Malik, H.S.; Anwar, M.

    2004-01-01

    Objective: To evaluate three different enzyme immunoassays for serological diagnosis of pulmonary tuberculosis and to compare their diagnostic accuracy in different combinations. Subjects and Methods: Sera from patients suffering from pulmonary tuberculosis (n=94) with sputum positive for acid fast bacilli (AFB) and sera from control group of healthy individuals (n=90) with sputum negative for AFB were tested by Pathozyme-Myco G EIA, Pathozyme-TB Complex Plus EIA and Pathozyme Myco M EIA kits for the genus-specific IgG and IgM, and the species-specific IgG antibodies against antigens of Mycobacterium tuberculosis. Results: The detection of IgG against genus-specific antigens by Pathozyme-Myco G had a sensitivity of 46% and a specificity of 93%, of IgG against species-specific antigens by Pathozyme- TB Complex Plus had a sensitivity of 64% and specificity of 97% and of IgM against genus-specific antigens by Pathozyme Myco M had a sensitivity of 67% and specificity of 98%. When the results of these immunoassays were evaluated in combination, their sensitivity improved. Combination of genus-specific IgM and species-specific IgG yielded best results with a sensitivity of 87% and specificity of 93%. Conclusion: The sensitivity of serological diagnosis of tuberculosis is low, but it can be increased by utilizing a combination of several antigens. (author)

  8. Biotin IgM Antibodies in Human Blood: A Previously Unknown Factor Eliciting False Results in Biotinylation-Based Immunoassays

    Science.gov (United States)

    Chen, Tingting; Hedman, Lea; Mattila, Petri S.; Jartti, Laura; Jartti, Tuomas; Ruuskanen, Olli; Söderlund-Venermo, Maria; Hedman, Klaus

    2012-01-01

    Biotin is an essential vitamin that binds streptavidin or avidin with high affinity and specificity. As biotin is a small molecule that can be linked to proteins without affecting their biological activity, biotinylation is applied widely in biochemical assays. In our laboratory, IgM enzyme immuno assays (EIAs) of µ-capture format have been set up against many viruses, using as antigen biotinylated virus like particles (VLPs) detected by horseradish peroxidase-conjugated streptavidin. We recently encountered one serum sample reacting with the biotinylated VLP but not with the unbiotinylated one, suggesting in human sera the occurrence of biotin-reactive antibodies. In the present study, we search the general population (612 serum samples from adults and 678 from children) for IgM antibodies reactive with biotin and develop an indirect EIA for quantification of their levels and assessment of their seroprevalence. These IgM antibodies were present in 3% adults regardless of age, but were rarely found in children. The adverse effects of the biotin IgM on biotinylation-based immunoassays were assessed, including four inhouse and one commercial virus IgM EIAs, showing that biotin IgM do cause false positivities. The biotin can not bind IgM and streptavidin or avidin simultaneously, suggesting that these biotin-interactive compounds compete for the common binding site. In competitive inhibition assays, the affinities of biotin IgM antibodies ranged from 2.1×10−3 to 1.7×10−4 mol/L. This is the first report on biotin antibodies found in humans, providing new information on biotinylation-based immunoassays as well as new insights into the biomedical effects of vitamins. PMID:22879954

  9. Specific Anti Mumps Antibodies (IgG & IgM in Cerebrospinal Fluid of Mumps Meningoencephalitic Children

    Directory of Open Access Journals (Sweden)

    Noorbakhsh Samileh

    2009-10-01

    Full Text Available Mumps infection is endemic in Iran. Our objective was to evaluate the presence of anti mumps antibodies ( IgM & IgG in cerebrospinal fluid in mumps meningoencephalitic children. A prospective/cross-sectional study was performed in Tehran, Iran (2003 to 2004 and serum anti mumps antibodies (IgM were detected (quantitive; ELISA in meningoencephaltis patients. Specific anti mumps antibodies (IgM & IgG were detected in cerebrospinal fluids of mumps meningoencephalitis cases. 43 meningoencephalitic patients were tested (59.2% male and 40.8% female. The age of patients was 79.96 ± 4.7 month. 23 (78.7% cases had specific mumps IgM in serum. None of cases had IgM antibodies in CSF. Anti mumps IgG antibody was detected in CSF of 7.5% (2/23 cases. We detected lower than expected frequency of local immunity to mumps virus in CSF of our cases. For better serologic diagnosis we recommend more sensitive methods like virus detection (PCR or short-term culture of lymphocytes from cerebrospinal fluid in future studies.

  10. [Prevalence of seropositividad to antibodies IgG and IgM against Helicobacter pylori in the medical residents of the University Hospital of Puebla].

    Science.gov (United States)

    Garza Yado, María de los Angeles; López García, Aída Inés; Paz Martínez, David; Galindo García, José Arturo; Cuevas Acuña, María Tula; Papaqui Tapia, Sergio; Arana Muñoz, Oswaldo; Pérez Fernández, María Susana

    2006-01-01

    Helicobacter pylori is a gram-negative, curved, microaerophilic, oxidase, catalase and urease positive bacillus. It lives in the gastric mucosa, and is the main etiological agent of peptic ulcer, and of atrophic and multifocal gastritis. It is associated with extraintestinal, vascular, autoimmune and cutaneous diseases. The infection by this bacteria causes a chronic inflammatory process related with the sensibilization of mast cells, which increases the incidence of allergic diseases. To estimate the seropositivity prevalence of IgG and IgM antibodies against Helicobacter pylori in medical resident personnel. A transversal, descriptive and observational study was carried out in medical residents of the Hospital Universitario de Puebla. Serum levels of IgG and IgM antibodies against Helicobacter pylori were determined by chemiluminescence. Data were analyzed applying descriptive statistics, such as: frequency, central tendency and dispersion measures. 57 residents were included, 54.4% were men. The mean age was 28.4 (SD 2.9). Seropositivity prevalence of IgG and IgM antibodies was 24.6 and 33.3%, respectively. Combined seropositivity prevalence of IgG and IgM was of 43.9%. Seropositivity prevalence of IgG and IgM antibodies against Helicobacter pylori in the medical resident personnel of the Hospital Universitario de Puebla was similar to the reported in the general population. This estimation might be considered a reference to other epidemiological and clinical studies such as those pretending to evaluate and demonstrate the relationship between infection by Helicobacter pylori and other entities, even allergies.

  11. In-Depth Analysis of Human Neonatal and Adult IgM Antibody Repertoires

    Directory of Open Access Journals (Sweden)

    Binbin Hong

    2018-02-01

    Full Text Available Although high-throughput sequencing and associated bioinformatics technologies have enabled the in-depth, sequence-based characterization of human immune repertoires, only a few studies on a relatively small number of sequences explored the characteristics of antibody repertoires in neonates, with contradictory conclusions. To gain a more comprehensive understanding of the human IgM antibody repertoire, we performed Illumina sequencing and IMGT/HighV-QUEST analysis of IgM heavy chain repertoire of the B lymphocytes from the cord blood (CB of neonates, as well as the repertoire from peripheral blood of healthy human adults (HH. The comparative study revealed unexpectedly high levels of similarity between the neonatal and adult repertoires. In both repertoires, the VDJ gene usage showed no significant difference, and the most frequently used VDJ gene was IGHV4-59, IGHD3-10, and IGHJ3. The average amino acid (aa length of CDR1 (CB: 8.5, HH: 8.4 and CDR2 (CB: 7.6, HH: 7.5, as well as the aa composition and the average hydrophobicity of the CDR3 demonstrated no significant difference between the two repertories. However, the average aa length of CDR3 was longer in the HH repertoire than the CB repertoire (CB: 14.5, HH: 15.5. Besides, the frequencies of aa mutations in CDR1 (CB: 19.33%, HH: 25.84% and CDR2 (CB: 9.26%, HH: 17.82% were higher in the HH repertoire compared to the CB repertoire. Interestingly, the most prominent difference between the two repertoires was the occurrence of N2 addition (CB: 64.87%, HH: 85.69%, a process that occurs during V-D-J recombination for introducing random nucleotide additions between D- and J-gene segments. The antibody repertoire of healthy adults was more diverse than that of neonates largely due to the higher occurrence of N2 addition. These findings may lead to a better understanding of antibody development and evolution pathways and may have potential practical value for facilitating the generation of more

  12. In-Depth Analysis of Human Neonatal and Adult IgM Antibody Repertoires

    Science.gov (United States)

    Hong, Binbin; Wu, Yanling; Li, Wei; Wang, Xun; Wen, Yumei; Jiang, Shibo; Dimitrov, Dimiter S.; Ying, Tianlei

    2018-01-01

    Although high-throughput sequencing and associated bioinformatics technologies have enabled the in-depth, sequence-based characterization of human immune repertoires, only a few studies on a relatively small number of sequences explored the characteristics of antibody repertoires in neonates, with contradictory conclusions. To gain a more comprehensive understanding of the human IgM antibody repertoire, we performed Illumina sequencing and IMGT/HighV-QUEST analysis of IgM heavy chain repertoire of the B lymphocytes from the cord blood (CB) of neonates, as well as the repertoire from peripheral blood of healthy human adults (HH). The comparative study revealed unexpectedly high levels of similarity between the neonatal and adult repertoires. In both repertoires, the VDJ gene usage showed no significant difference, and the most frequently used VDJ gene was IGHV4-59, IGHD3-10, and IGHJ3. The average amino acid (aa) length of CDR1 (CB: 8.5, HH: 8.4) and CDR2 (CB: 7.6, HH: 7.5), as well as the aa composition and the average hydrophobicity of the CDR3 demonstrated no significant difference between the two repertories. However, the average aa length of CDR3 was longer in the HH repertoire than the CB repertoire (CB: 14.5, HH: 15.5). Besides, the frequencies of aa mutations in CDR1 (CB: 19.33%, HH: 25.84%) and CDR2 (CB: 9.26%, HH: 17.82%) were higher in the HH repertoire compared to the CB repertoire. Interestingly, the most prominent difference between the two repertoires was the occurrence of N2 addition (CB: 64.87%, HH: 85.69%), a process that occurs during V-D-J recombination for introducing random nucleotide additions between D- and J-gene segments. The antibody repertoire of healthy adults was more diverse than that of neonates largely due to the higher occurrence of N2 addition. These findings may lead to a better understanding of antibody development and evolution pathways and may have potential practical value for facilitating the generation of more effective

  13. Isolation and Characterization of IgM and IgY Antibodies from Plasma of Magellanic Penguins (Spheniscus magellanicus).

    Science.gov (United States)

    Bizelli, Camila C; Silva, A Sandriana R; da Costa, Jessica D; Vanstreels, Ralph E T; Atzingen, Marina V; Santoro, Marcelo L; Fernandes, Irene; Catão-Dias, José L; Faquim-Mauro, Eliana L

    2015-03-01

    Infectious diseases such as aspergillosis, avian malaria, and viral infections are significant threats to the conservation of penguins, leading to morbidity and mortality of these birds both in captivity and in the wild. The immune response to such infectious diseases is dependent on different mechanisms mediated by cells and soluble components such as antibodies. Antibodies or immunoglobulins are glycoproteins that have many structural and functional features that mediate distinct effector immune functions. Three distinct classes of antibodies have been identified in birds: immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin Y (IgY). In this study we aim to establish an efficient laboratory method to obtain IgM and IgY antibodies from plasma samples of healthy adult Magellanic penguins (Spheniscus magellanicus). The protocol was developed combining plasma delipidation, sequential precipitation with caprylic acid and ammonium sulfate, and size-exclusion chromatography. The efficiency of the protocol and the identity of the purified IgM and IgY antibodies were confirmed through enzyme-linked immunosorbent assay, Western blotting, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, and lectin binding assay. Structural and physicochemical properties of IgM and IgY from Magellanic penguins were consistent with those of other avian species. This purification protocol will allow for more detailed studies on the humoral immunity of penguins and for the development of high specificity serologic assays to test Magellanic penguins for infectious pathogens.

  14. Serologic cross-reactivity of human IgM and IgG antibodies to five species of Ebola virus.

    Directory of Open Access Journals (Sweden)

    Adam Macneil

    2011-06-01

    Full Text Available Five species of Ebola virus (EBOV have been identified, with nucleotide differences of 30-45% between species. Four of these species have been shown to cause Ebola hemorrhagic fever (EHF in humans and a fifth species (Reston ebolavirus is capable of causing a similar disease in non-human primates. While examining potential serologic cross-reactivity between EBOV species is important for diagnostic assays as well as putative vaccines, the nature of cross-reactive antibodies following EBOV infection has not been thoroughly characterized. In order to examine cross-reactivity of human serologic responses to EBOV, we developed antigen preparations for all five EBOV species, and compared serologic responses by IgM capture and IgG enzyme-linked immunosorbent assay (ELISA in groups of convalescent diagnostic sera from outbreaks in Kikwit, Democratic Republic of Congo (n=24, Gulu, Uganda (n=20, Bundibugyo, Uganda (n=33, and the Philippines (n=18, which represent outbreaks due to four different EBOV species. For groups of samples from Kikwit, Gulu, and Bundibugyo, some limited IgM cross-reactivity was noted between heterologous sera-antigen pairs, however, IgM responses were largely stronger against autologous antigen. In some instances IgG responses were higher to autologous antigen than heterologous antigen, however, in contrast to IgM responses, we observed strong cross-reactive IgG antibody responses to heterologous antigens among all sets of samples. Finally, we examined autologous IgM and IgG antibody levels, relative to time following EHF onset, and observed early peaking and declining IgM antibody levels (by 80 days and early development and persistence of IgG antibodies among all samples, implying a consistent pattern of antibody kinetics, regardless of EBOV species. Our findings demonstrate limited cross-reactivity of IgM antibodies to EBOV, however, the stronger tendency for cross-reactive IgG antibody responses can largely circumvent

  15. Level of reactivity of IgM anticardiolipin antibodies in preeclampsia: a likely early serologic marker

    Directory of Open Access Journals (Sweden)

    Karla N. Pereira

    2015-02-01

    Full Text Available The purpose of this study was to measure the level of anticardiolipin antibodies in preeclamptic pregnant women to verify a possible association between the presence of these antibodies and the development of the syndrome. A total of 36 pregnant women with preeclampsia and 19 women with normal pregnancy were evaluated. Anticardiolipin antibodies were determined by enzyme immunoassay. The reactivity level of anticardiolipin antibodies was significantly higher in the preeclamptic group, with a significance level of 95%. The results can mean the participation of anticardiolipin antibodies in the pathogenesis of preeclampsia. Future studies may confirm this parameter as an early marker for the development of this disease.

  16. Anti-N-methyl-D-aspartate receptor encephalitis with positive serum antithyroid antibodies, IgM antibodies against mycoplasma pneumoniae and human herpesvirus 7 PCR in the CSF.

    Science.gov (United States)

    Venâncio, Paulo; Brito, Maria João; Pereira, Gabriela; Vieira, José Pedro

    2014-08-01

    We report the case of a boy with an encephalopathy associated with extrapyramidal and psychiatric symptoms and anti-N-methyl-D-aspartate receptor antibodies. He had positive serum antithyroid antibodies, IgM antibodies against Mycoplasma pneumoniae and human herpesvirus 7 polymerase chain reaction in the cerebrospinal fluid. He was successfully treated with rituximab, after steroids, intravenous immunoglobulin and plasma exchange. The pathophysiology of this disorder may be post-infectious and autoimmune.

  17. A comparison of antibody capture radio- and enzyme immunoassays with immunofluorescence for detecting IgM antibody in infants with congenital rubella

    International Nuclear Information System (INIS)

    Chantler, S.; Evans, C.J.; Mortimer, P.P.; Cradock-Watson, J.E.; Ridehalgh, M.K.S.

    1982-01-01

    IgM antibody capture radioimmunoassay (MACRIA) and enzyme immunoassay (MACEIA) were compared with immunofluorescence (IF) for detecting specific IgM antibody in 99 sera from 76 infants with confirmed congenital rubella, and 61 sera from a comparative group of 59 infants who had miscellaneous abnormalities but in whom congenital rubella was not confirmed. All of 35 specimens collected from confirmed cases within 12 weeks of birth were positive by all three methods and all but one of 17 specimens collected after the age of 18 months were uniformly negative. At intermediate ages discrepancies occurred in 18 specimens, of which eight were positive and 10 negative by IF. Three of these 18 specimens were negative by both antibody capture procedures but showed weak fluorescence; the other 15 were negative by MACEIA, but positive by MACRIA which appears to be the more sensitive of the antibody capture methods. Sera from five infants in the comparative group were clearly positive by all three methods. These five infants were probably congenitally infected with rubella. Sera from the other 54 infants were negative, except for one that gave a weakly positive result by MACRIA alone. Antibody capture procedures offer several advantages over previous methods for detecting IgM antibody. Although MACRIA was found to be slightly more sensitive than MACEIA, the greater stability of the enzyme label, together with the possibility of both visual and quantitative assessment, could make MACEIA the method of choice for detecting rubella-specific IgM. (Auth.)

  18. Comparison of antibody capture radio- and enzyme immunoassays with immunofluorescence for detecting IgM antibody in infants with congenital rubella

    Energy Technology Data Exchange (ETDEWEB)

    Chantler, S.; Evans, C.J. (Wellcome Research Lab., Beckenham (UK)); Mortimer, P.P. (Virus Reference Laboratory, Central Public Health Laboratory, Colindale Avenue, London, (UK)); Cradock-Watson, J.E.; Ridehalgh, M.K.S. (Withington Hospital, Manchester (UK))

    1982-08-01

    IgM antibody capture radioimmunoassay (MACRIA) and enzyme immunoassay (MACEIA) were compared with immunofluorescence (IF) for detecting specific IgM antibody in 99 sera from 76 infants with confirmed congenital rubella, and 61 sera from a comparative group of 59 infants who had miscellaneous abnormalities but in whom congenital rubella was not confirmed. All of 35 specimens collected from confirmed cases within 12 weeks of birth were positive by all three methods and all but one of 17 specimens collected after the age of 18 months were uniformly negative. At intermediate ages discrepancies occurred in 18 specimens, of which eight were positive and 10 negative by IF. Three of these 18 specimens were negative by both antibody capture procedures but showed weak fluorescence; the other 15 were negative by MACEIA, but positive by MACRIA which appears to be the more sensitive of the antibody capture methods. Sera from five infants in the comparative group were clearly positive by all three methods. These five infants were probably congenitally infected with rubella. Sera from the other 54 infants were negative, except for one that gave a weakly positive result by MACRIA alone. Antibody capture procedures offer several advantages over previous methods for detecting IgM antibody. Although MACRIA was found to be slightly more sensitive than MACEIA, the greater stability of the enzyme label, together with the possibility of both visual and quantitative assessment, could make MACEIA the method of choice for detecting rubella-specific IgM.

  19. Diagnostic value of specific IgM antibodies in primary HIV infection

    NARCIS (Netherlands)

    Lange, J. M.; Parry, J. V.; de Wolf, F.; Mortimer, P. P.; Goudsmit, J.

    1988-01-01

    Sequential serum samples from 55 homosexual men with primary HIV infection were tested for IgM anti-HIV. An early IgM response was found in 27 out of 55 (49%). In five cases IgM anti-HIV was detected 1-3 1/4 months prior to IgG anti-HIV seroconversion, as detected by a commercially available ELISA,

  20. Atheroprotective natural anti-phosphorylcholine antibodies of IgM subclass are decreased in Swedish controls as compared to non-westernized individuals from New Guinea

    Directory of Open Access Journals (Sweden)

    Råstam Lennart

    2007-03-01

    Full Text Available Abstract Objective To determine the importance of IgM antibodies against phosphorylcholine (aPC, a novel protective factor for cardiovascular disease (CVD, in a population with a non-western life style as compared with a Swedish control group. Methods and results Risk factors for cardiovascular disease were determined in a group of 108 individuals aged 40–86 years from New Guinea and 108 age-and sex-matched individuals from a population based study in Sweden. Antibodies were tested by ELISA. aPC IgM levels were significantly higher among New Guineans than among Swedish controls (p Conclusion IgM-antibodies against PC, which have atheroprotective properties, are higher in a population from Kitava, New Guinea with a traditional lifestyle, than in Swedish Controls, and higher among women than men in both populations tested. Such antibodies could contribute to the low incidence of cardiovascular disease reported from Kitava and could also provide an explanation as to why women have a later onset of CVD than men.

  1. Toxocara canis glycans influence antigen recognition by mouse IgG1 and IgM antibodies.

    Science.gov (United States)

    Długosz, Ewa; Wiśniewski, Marcin

    2016-01-01

    The impact of sugar moieties of Toxocara canis glycoprotein antigens on their recognition by infected mouse antibodies was investigated in this study. Native TES and recombinant Toxocara mucins generated in Pichia pastoris yeast as well as their deglycosylated forms were used in ELISA. TES and recombinant mucins were equally recognized by T. canis infected mouse IgG1 antibodies. IgM immunoglobulins predominantly recognized TES antigens. Among mucins recognition of Tc-MUC-4 was the most significant. Deglycosylation of antigens resulted in significant loss of IgM and IgG1 reactivity to TES, mucins, Tc-MUC-3 and Tc-MUC-4. The presence of sugar moieties had no influence on IgE binding to native or recombinant T. canis antigens. Our results suggest that glycans are involved in epitope formation what should be taken into consideration in production of recombinant helminth antigens for diagnostic purposes.

  2. Association of beta2-glycoprotein I IgG and IgM antibodies with thrombosis and thrombocytopenia

    DEFF Research Database (Denmark)

    Voss, Anne-Sofie Boertmann; Jacobsen, Søren; Heegaard, Niels Henrik Helweg

    2001-01-01

    Antiphospholipid antibodies (APA) have been known for decades. Their relation to clinical manifestations, primarily thromboses and thrombocytopenia, was recognised in the 1980s. In this clinical study two cohorts of patients, a population-based (84 patients with systemic lupus erythematosus (SLE......)) and a hospital-based (87 patients with SLE and 53 with other connective tissue diseases) were investigated for APA and associated clinical manifestations. Anticardiolipin antibodies (ACA) of IgG and IgM classes were found in 13 and 38% of the population-based patients and in 29 and 58% of the hospital...

  3. IgM-class rheumatoid factor interference in the solid-phase radioimmunoassay of rubella-specific IgM antibodies

    International Nuclear Information System (INIS)

    Meurman, O.H.; Ziola, B.R.

    1978-01-01

    The interference of IgM-class rheumatoid factor (RF) in the solid-phase radioimmunoassay (RIA) of rubella virus IgM antibodies was studied. Acute rubella infections did not significantly activate RF. False-positive rubella antibody results were obtained, however, when patients with raised RF levels were tested. If a low rubella IgG antibody titre was present, a high level of RF was required to cause a false-positive IgM result; conversely, in sera with high IgG titres, only a low level of RF was required for interference. Although the false-positive IgM titres obtained were generally low, they did show a positive correlation to both RF levels and rubella IgG titres. False-positive results were successfully avoided by removing the RF by absorption with heat-aggregated human gamma globulin. The absorption procedure did not affect true rubella IgM antibody titres. (author)

  4. Detection of IgM antibodies against Toxoplasma gondii in blood samples absorbed onto filter paper, in order to assess a baseline for the occurence of congenital toxoplasma infections

    NARCIS (Netherlands)

    Conyn-van Spaendonck MAE; van Knapen F

    1990-01-01

    This report deals in the first place with some investigations on the utility of blood samples absorbed onto filter paper for determination of antibodies to Toxoplasma gondii. Secondly, the results of specific IgM detection in such samples of a cohort of neonates born in 1986/1987 are reported. The

  5. IgM but not IgG monoclonal anti-Nocardia brasiliensis antibodies confer protection against experimental actinomycetoma in BALB/c mice.

    Science.gov (United States)

    Gonzalez-Suarez, Maria L; Salinas-Carmona, Mario C; Pérez-Rivera, Isabel

    2009-10-01

    Nocardia brasiliensis is a facultative intracellular microorganism that produces a human chronic infection known as actinomycetoma. Human and mouse anti-N. brasiliensis antibody response identify P24, P26 and P61 immunodominant antigens. In this work, we generated immunoglobulin M (IgM) and IgG monoclonal antibodies (mAbs) specific to immunodominant P61 antigen. The monoclonal IgM (NbM1) and IgG2a (NbG1) antibodies were assessed for their in vitro bactericidal activity, in vivo protective effect and ability to block catalase activity. These mAbs specifically recognized P61, but they did not inhibit its enzyme activity. The in vitro bactericidal effect of NbG1 was higher than the killing ability of the IgM mAb. In vivo experiments with a murine model of experimental infection with N. brasiliensis injected into rear footpads was used to test the effect of NbM1 and NbG1. The negative untreated group developed a chronic actinomycetoma within 4 weeks. IgM mAbs conferred protection to BALB/c mice infected with N. brasiliensis. IgG mAb lacked this protective effect. IgM mAb showed a dose-response correlation between antibody concentration and lesion size. These results demonstrate that humoral immune response mediated by antigen-specific IgM antibody protects against an intracellular bacterial infection.

  6. ¿Es necesario investigar anticuerpos IgM contra el virus de la hepatitis A cuando el enzimograma hepático es normal? Is it necessary to investigate anti-hepatitis A virus (HAV IgM antibodies when the hepatic enzymogram is normal?

    Directory of Open Access Journals (Sweden)

    C. Coitinho

    2007-09-01

    levels of serum aspartate and alanine aminotransferases (AST/ALT. All patients (n = 158 received in the laboratory requesting a hepatic enzymograme and anti-HAV IgM were evaluated in a prospective study between October 2005 and March 2006. Anti-HAV IgM assays were carried out by microparticle enzyme immunoassay (MEIA. The quantification of hepatic enzymes was made in a multianalyzer. The most frequent clinical data were: presumption of hepatitis and jaundice (27.5 and 12.7%. Eighty four of the 158 patients (53% showed elevated values of ALT and AST, whereas 69 patients in this group (82% were anti-Hav IgM reactive. The remaining 74 patients (47% showed normal levels of AST/ALT and none of them were anti-HAV IgM reactive, except 7, who were on control of a confirmed HAV infection. Of the anti-HAV IgM reactive group of patientss, 49% were children under 10 years of age. Laboratory HAV confirmatory tests would have to be made in sequential form, the determination of anti-HAV IgM antibodies being unnecessary when normal values of serum aminotransferases are observed.

  7. IgM antibody and colony fungal load impacts of orally administered ethanol extract of Plectranthus scutellarioides on mice with systemic candidiasis

    Directory of Open Access Journals (Sweden)

    Marni Br. Karo

    2018-02-01

    Full Text Available Context: Candida albicans is a pathogenic fungus that causes vulvovaginal candidiasis (VVC, which presents significant clinical problems in immunocompromised patients. There are no studies that correlate IgM antibody responses and fungal loads following the use of ethanol leaf extract of Plectranthus scutellarioides (L. R.Br. (ELEP on VVC. Aims: To examine colony fungi loads, IgM antibody responses, and their correlation in Balb/c mice infected with C. albicans treated with orally administered ELEP. Methods: Twenty-five female Balb/c mice were randomly divided into five groups (five mice in each group: normal group (oral ELEP + not infected, VVC control (VCC group (infected + oral 0.2 mL saline, ketoconazole-treated in VCC group (infected + oral antifungal agents, and two treatment groups (infected + oral ELEP with 500 and 750 mg/kg b.w.. Treatments were administered for 14 days after infection with C. albicans. Colony fungal loads and IgM antibodies were measured by counting colony forming units and flow cytometry using anti-C. albicans antibody kits, respectively. Results: Ingestion of the ELEP significantly affected fungal load clearance and decreased IgM antibody levels compared to that in the VCC group (p<0.001. Furthermore, ingestion of the ELEP was associated with significantly lower concentrations of IgM antibodies and colony forming units (p<0.001. Conclusions: These data strongly suggest that IgM antibody response is involved in of the mechanism by which ELEP decreases C. albicans infection. Furthermore, these results provide important information for the potential application of ELEP in the treatment of Balb/C mice with systemic candidiasis.

  8. Validation of an IgM antibody capture ELISA based on a recombinant nucleoprotein for identification of domestic ruminants infected with Rift Valley fever virus.

    Science.gov (United States)

    Williams, Roy; Ellis, Charlotte Elizabeth; Smith, Shirley Jacqueline; Potgieter, Christiaan Abraham; Wallace, David; Mareledwane, Vuyokazi Epipodia; Majiwa, Phelix Antipas Ochola

    2011-11-01

    The presence of competent vectors in some countries currently free of Rift Valley fever (RVF) and global changes in climate, travel and trade have increased the risk of RVF spreading to new regions and have emphasised the need for accurate and reliable diagnostic tools for early diagnosis during RVF outbreaks. Highly sensitive viral detection systems like PCR have a limited use during outbreaks because of the short duration of viraemia, whereas antibodies like specific IgM which are serological indicators of acute infection, can be detected for up to 50 days after infection. Using the highly conserved and immunogenic recombinant nucleoprotein of RVF virus in an IgM capture ELISA, the risk of laboratory infection associated with traditional serological methods is avoided. The use of pre-coated/pre-blocked ELISA plates and the conjugation of the recombinant nucleoprotein with horseradish peroxidase simplified and shortened the assay procedure. Results showed the assay to be highly reproducible with a lower detection limit equal to that of a commercial competition ELISA. By receiver operating characteristic (ROC) curve analysis the area under curve (AUC) index was determined as 1.0 and the diagnostic sensitivity and specificity at a PP cut-off value of 4.1 as 100% and 99.78% respectively. The results of this study demonstrated that the IgM capture ELISA is a safe, reliable and highly accurate diagnostic tool which can be used on its own or in parallel with other methods for the early diagnosis of RVF virus infection and also for monitoring of immune responses in vaccinated domestic ruminants. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. A COMPARATIVE EPIDEMIOLOGIC STUDY OF SPECIFIC ANTIBODIES (IgM AND IgA AND PARASITOLOGICAL FINDINGS IN AN ENDEMIC AREA OF LOW TRANSMISSION OF Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    KANAMURA Herminia Yohko

    1998-01-01

    Full Text Available The diagnostic potential of circulating IgM and IgA antibodies against Schistosoma mansoni gut-associated antigens detected by the immunofluorescence test (IFT on adult worm paraffin sections was evaluated comparatively to the fecal parasitological method, for epidemiological purposes in low endemic areas for schistosomiasis. Blood samples were collected on filter paper from two groups of schoolchildren living in two different localities of the municipality of Itariri (São Paulo, Brazil with different histories and prevalences of schistosomiasis. The parasitological and serological data were compared to those obtained for another group of schoolchildren from a non-endemic area for schistosomiasis. The results showed poor sensitivity of the parasitological method in detecting individuals with low worm burden and indicate the potential of the serological method as an important tool to be incorporated into schistosomiasis control and vigilance programs for determining the real situation of schistosomiasis in low endemic areas.

  10. Genetic parameters of IgM and IgG antibodies binding autoantigens in healthy chickens

    NARCIS (Netherlands)

    Bao, Mandy; Bovenhuis, H.; Nieuwland, M.G.B.; Parmentier, H.K.; Poel, van der J.J.

    2016-01-01

    Levels of natural antibodies (NAb) binding keyhole limpet hemocyanin (KLH) in layers were shown to be heritable and to be potential indicative parameters for survival. A proportion of NAb are directed to self-molecules, or slightly changed self-molecules (neo-epitopes), labeled as natural

  11. Detection of IgM antibodies against canine distemper virus in dog and mink sera employing enzyme-linked immunosorbent assay (ELISA).

    Science.gov (United States)

    Blixenkrone-Møller, M; Pedersen, I R; Appel, M J; Griot, C

    1991-01-01

    An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against canine distemper virus (CDV) in canine and mink serum is described. The diagnostic potential of this technique was evaluated by analyzing sera from natural or experimental infections in dog and mink and negative control sera. These results were compared with results obtained in the developed CDV IgG ELISA and in the virus neutralization test. The IgM test, which requires only a single serum specimen, is a useful method for diagnosing current or recent CDV infections in dog and mink.

  12. [Nephelometry or turbidimetry for the determination of albumin, ApoA, CRP, haptoglobin, IgM and transthyretin: which choice?].

    Science.gov (United States)

    Thuillier, F; Demarquilly, C; Szymanowicz, A; Gaillard, C; Boniface, M; Braidy, C; Daunizeau, A; Gascht, D; Gruson, A; Lagabrielle, J-F; Lasnier, E; Lemdani, M; Macchi, V; Poulin, G; de Sainte Hermine, C; Sancho, J; Schellenberg, F; Sevin, O; Vaubourdolle, M

    2008-01-01

    Nephelometry, which is considered as the reference method for serum proteins determination requires a specific equipment. The majority of protein determinations are therefore carried out on biochemistry automats using turbidimetry. The objective of a CNBH group (Collège national de biochimie des hôpitaux) was to compare nephelometry and turbidimetry for 7 automats: 2 nephelometers, the BN Prospec (Dade-Behring) and Immage (Beckman-Coulter) and 5 biochemistry systems using turbidimetry, the Integra and Modular (Roche Diagnostics), the LX20 (Beckman-Coulter), RXL (Dade-Behring) and AU (Olympus). The study was based on the determination of sera collections (albumin, ApoA, CRP, haptoglobin, IgM, transthyretin) of 140 samples each: 110 limpid samples and 30 samples called HLI (hemolytic, lipemic or icteric). Fifteen hospitals took part to this work. An ANOVA analysis on limpid samples and quality control sera concluded to an "automat" effect for the 6 tested proteins but did not show a "method" effect, (i.e. nephelometry versus turbidimetry). On the other hand, the transferability of the results was expected to be better and an effort on the choice of the antibodies and the standardization procedures should be made.

  13. High-Throughput Sequencing of the Expressed Torafugu (Takifugu rubripes) Antibody Sequences Distinguishes IgM and IgT Repertoires and Reveals Evidence of Convergent Evolution.

    Science.gov (United States)

    Fu, Xi; Sun, Jianqiang; Tan, Engkong; Shimizu, Kentaro; Reza, Md Shaheed; Watabe, Shugo; Asakawa, Shuichi

    2018-01-01

    B-cell antigen receptor (BCR) or antibody diversity arises from somatic recombination of immunoglobulin (Ig) gene segments and is concentrated within the Ig heavy (H) chain complementarity-determining region 3 (CDR-H3). We performed high-throughput sequencing of the expressed antibody heavy-chain repertoire from adult torafugu. We found that torafugu use between 70 and 82% of all possible V (variable), D (diversity), and J (joining) gene segment combinations and that they share a similar frequency distribution of these VDJ combinations. The CDR-H3 sequence repertoire observed in individuals is biased with the preferential use of a small number of VDJ, dominated by sequences containing inserted nucleotides. We uncovered the common CDR-H3 amino-acid (aa) sequences shared by individuals. Common CDR-H3 sequences feature highly convergent nucleic-acid recombination compared with private ones. Finally, we observed differences in repertoires between IgM and IgT, including the unequal usage frequencies of V gene segment and the biased number of nucleotide insertion/deletion at VDJ junction regions that leads to distinct distributions of CDR-H3 lengths.

  14. High-Throughput Sequencing of the Expressed Torafugu (Takifugu rubripes Antibody Sequences Distinguishes IgM and IgT Repertoires and Reveals Evidence of Convergent Evolution

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    Xi Fu

    2018-02-01

    Full Text Available B-cell antigen receptor (BCR or antibody diversity arises from somatic recombination of immunoglobulin (Ig gene segments and is concentrated within the Ig heavy (H chain complementarity-determining region 3 (CDR-H3. We performed high-throughput sequencing of the expressed antibody heavy-chain repertoire from adult torafugu. We found that torafugu use between 70 and 82% of all possible V (variable, D (diversity, and J (joining gene segment combinations and that they share a similar frequency distribution of these VDJ combinations. The CDR-H3 sequence repertoire observed in individuals is biased with the preferential use of a small number of VDJ, dominated by sequences containing inserted nucleotides. We uncovered the common CDR-H3 amino-acid (aa sequences shared by individuals. Common CDR-H3 sequences feature highly convergent nucleic-acid recombination compared with private ones. Finally, we observed differences in repertoires between IgM and IgT, including the unequal usage frequencies of V gene segment and the biased number of nucleotide insertion/deletion at VDJ junction regions that leads to distinct distributions of CDR-H3 lengths.

  15. Prevalence of anti-Rift-Valley-fever IgM antibody in abattoir workers in the Nile delta during the 1993 outbreak in Egypt.

    Science.gov (United States)

    Abu-Elyazeed, R.; el-Sharkawy, S.; Olson, J.; Botros, B.; Soliman, A.; Salib, A.; Cummings, C.; Arthur, R.

    1996-01-01

    In the early summer of 1993, an outbreak of Rift Valley fever (RVF) was reported among both humans and animals in Aswan Governorate, Upper Egypt. To determine whether RVF infection had spread to the Nile delta region of the country, we carried out a cross-sectional survey of 1181 occupationally exposed abattoir workers (97% male; age 10-72 years) in 15 governorates of Egypt in November 1993. The overall prevalence of anti-RVF virus IgM antibody was 2% (range: 0% (7 governorates) to 10%). The highest prevalences were in Ismailia (10%) and Sharqîya (8%) Governorates. None of the seropositive subjects reported having experienced an episode of fever in the 2 months prior to the study. The prevalence of antibody was significantly higher (P < 0.05) among workers employed in high-risk jobs such as cutting animals' throats (relative risk (RR = 2.24)) and handling animal parts (RR = 2.37). The findings suggest that abattoir workers represent a useful sentinel population for surveillance of RVF. PMID:8706230

  16. Gold magnetic nanoparticle conjugate-based lateral flow assay for the detection of IgM class antibodies related to TORCH infections.

    Science.gov (United States)

    Li, Xingxing; Zhang, Qinlu; Hou, Peng; Chen, Mingwei; Hui, Wenli; Vermorken, Alphons; Luo, Zhiyi; Li, Hong; Li, Qin; Cui, Yali

    2015-11-01

    In this study, a lateral flow immunochromatographic assay (LFIA) system for the detection of immunoglobulin M (IgM) antibodies, related to TORCH [(T)oxoplasmosis, (O)ther agents, (R)ubella (also known as German Measles), (C)ytomegalovirus, and (H)erpes simplex virus infections], based on gold magnetic nanoparticles, was established. Following modification with poly(methacrylic acid), the gold magnetic nanoparticles conjugated with an anti‑human IgM antibody (μ‑chain specific) to construct a probe. A lateral flow assay device was constructed based on these conjugates. IgM antibodies to four types of pathogens, notably toxoplasmosis, rubella virus, cytomegalovirus and herpes simplex virus type 2, were detected using this device. Compared with commercial colloidal gold‑based LFIA strips, our method exhibited higher sensitivity. No interference with triglycerides, hemoglobin and bilirubin occurred, and no cross‑reactivity was noted among the four pathogens. The gold magnetic nanoparticle‑LFIA strips were used to assess 41 seropositive and 121 seronegative serum samples. The sensitivity was 100% (162/162) and the specificity was 100% (162/162). This method cannot only be used for the detection of TORCH IgM-specific antibodies, but it can potentially be developed for use in the diagnosis of other acute or recently identified autoimmune diseases.

  17. Duration of secretory IgM and IgA antibodies to respiratory syncytial virus in a community study in Guinea-Bissau

    DEFF Research Database (Denmark)

    Stensballe, L G; Kofoed, P E; Nante, E J

    2000-01-01

    nasopharyngeal suction on days 7, 14, 30, 60 and 120 in the first epidemic and every fortnight for 6 mo after the second epidemic to measure the duration of secretory IgM and IgA responses. Nearly all of the children had an IgM response during the first month after infection. The response ratio was highest...... phase of infection. A secondary response may be more likely in children with low IgM responses in the acute phase (RR = 2.08 (95% confidence interval (CI) 0.92-4.70)). The IgA response was highest on days 28 and 42 after antigen detection, 72% having a detectable IgA response within the first 1.5 mo...... have therefore assessed the duration of secretory IgM and IgA antibody responses and whether assays for these antibodies can be used to improve the diagnosing of RSV-associated infections. During two RSV epidemics in Guinea-Bissau, 32 RSV antigen-positive children with LRI were followed with sequential...

  18. IgM response to a human Pneumocystis carinii surface antigen in HIV-infected patients with pulmonary symptoms

    DEFF Research Database (Denmark)

    Lundgren, Bettina; Kovacs, J A; Mathiesen, Lars Reinhardt

    1993-01-01

    We have developed an ELISA to detect IgM antibodies to a major human Pneumocystis carinii surface antigen (gp95), and investigated the IgM response in 128 HIV-infected patients who underwent bronchoscopy for evaluation of pulmonary symptoms. Only 5 (4%) patients had IgM antibodies to P. carinii gp...... human P. carinii surface antigen. The increase in IgM response during the course of PCP observed in 3 patients suggests either reinfection with a new strain, or antigenic drift of an already acquired strain of P. carinii.......95. Four of the 5 patients with IgM antibodies also had IgG antibodies to gp95 and microbiologically proven P. carinii pneumonia (PCP). In 76/128 patients for whom serial samples were available, changes in antibody response were determined. In 3 patients we demonstrated an increase in IgM antibody...

  19. Hepatitis B Virus Infection and Hepatocellular Carcinoma: Correlation Between IgM Antibody to Hepatitis B Core Antigen, Hepatitis B e Antigen, and Hepatitis B DNA

    Science.gov (United States)

    1988-01-01

    Mdicine and ni)giene HEPATITIS B VIRUS INFECTION AND HEPATOCELLULAR CARCINOMA: CORRELATION BETWEEN IgM ANTIBODY TO HEPATITIS B CORE ANTIGEN, HEPATITIS B e...ANTIGEN, AND HEPATITIS B DNA MARIA H. SJOGREN.* GEOFFREY M. DUSHEIKO. MICHAEL C. KEW, AND ERNEST SONG *Depart,ent of Virus Diseases, valter Reed Arny...Johannesburg, South Africa Abstract. Sera from 102 black patients with primary hepatocellular carcinoma (PHC) and hepatitis B surface antigenemia

  20. Detection of IgM and IgG antibodies to Chlamydophila pneumoniae in pediatric community-acquired lower respiratory tract infections

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    Surinder Kumar

    2011-01-01

    Full Text Available Context: Chlamydophila pneumoniae (C. pneumoniae is an emerging infectious agent with a spectrum of clinical manifestations including lower and upper respiratory tract infections. Aims: To investigate the role of C. pneumoniae in community-acquired lower respiratory tract infections (LRTIs in children using serological tests. Settings and Design: Two hundred children, age 2 months to 12 years, hospitalized for community-acquired LRTIs were investigated for C. pneumoniae etiology. Materials and Methods: We investigated 200 children hospitalized for community-acquired LRTIs, using ELISA for detecting anti-C. pneumoniae IgM and IgG antibodies. The demographic, clinical and radiological findings for C. pneumoniae antibody positive and C. pneumoniae antibody negative cases were compared. Statistical Analysis Used: Data analysis was performed by Chi-square test and Fisher′s exact tests using Epi Info (2002. Results: Clinical and radiological findings in both the groups were comparable. Serological evidence of C. pneumoniae infection was observed in 12 (6% patients; specific IgM antibodies were detected in 11 (91.67%; specific IgG antibodies in 1 (8.33% patients, while 4-fold rise in C. pneumoniae IgG antibody titers were noted in none of the patients. Conclusions: C. pneumoniae has a role in community-acquired LRTIs, even in children aged < 5 years. Serological detection using ELISA would enable pediatricians in better management of C. pneumoniae infections.

  1. Natural IgM antibodies that bind neoepitopes exposed as a result of spinal cord injury , drive secondary injury by activating complement.

    Science.gov (United States)

    Narang, Aarti; Qiao, Fei; Atkinson, Carl; Zhu, Hong; Yang, Xiaofeng; Kulik, Liudmila; Holers, V Michael; Tomlinson, Stephen

    2017-06-19

    Natural IgM antibodies (Abs) function as innate immune sensors of injury via recognition of neoepitopes expressed on damaged cells, although how this recognition systems function following spinal cord injury (SCI) exposes various neoepitopes and their precise nature remains largely unknown. Here, we investigated the role of two natural IgM monoclonal Abs (mAbs), B4 and C2, that recognize post-ischemic neoepitopes following ischemia and reperfusion in other tissues. Identification of post-SCI expressed neoepitopes was examined using previously characterized monoclonal Abs (B4 and C2 mAbs). The role of post-SCI neoepitopes and their recognition by natural IgM Abs in propagating secondary injury was examined in Ab-deficient Rag1-/- or wild type C57BL/6 mice using Ab reconstitution experiments and neoepitope-targeted therapeutic studies, respectively. Administration of B4 or C2 mAb following murine SCI increased lesion size and worsened functional outcome in otherwise protected Ab-deficient Rag1-/- mice. Injury correlated with colocalized deposition of IgM and C3d in injured spinal cords from both mAb reconstituted Rag1-/- mice and untreated wild-type mice. Depletion of peritoneal B1 B cells, a source of natural Abs, reduced circulating levels of IgM with B4 (annexin-IV) and C2 (subset of phospholipids) reactivity, reduced IgM and complement deposition in the spinal cord, and protected against SCI. We therefore investigated whether the B4 neoepitope represents a therapeutic target for complement inhibition. B4-Crry, a fusion protein consisting of a single-chain Ab derived from B4 mAb, linked to the complement inhibitor Crry, significantly protected against SCI. B4-Crry exhibited a dual function in that it inhibited both the binding of pathogenic IgM and blocked complement activation in the spinal cord. This study identifies important neoepitopes expressed within the spinal cord after injury. These neoepitopes are recognized by clonally specific natural IgM Abs that

  2. Synthetic positive controls for ELISA test kits for detection of IgA and IgM antibodies to Chlamydia trachomatis

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    O. Y. Galkin

    2015-01-01

    Full Text Available The enzyme-linked immunosorbent assay (ELISA is the most informative and versatile method of serological diagnostics. The possibility of detecting by ELISA specific antibodies of different classes allow to differentiate primary infectious process and its remission, exacerbation and chronic disease (holding of differential diagnosis. This approach is implemented in the methodology for evaluation of patients for presence of humoral immune response against the causative agent of urogenital chlamydiosis. As with other infections immediately after Chlamydia trachomatis infection the specific IgM antibodies are formed, and subsequently basic projective antibodies of IgG class are synthesized. However, at exacerbation of chronic urogenital chlamydiosis specific IgA antibodies can be synthesized. That is why comprehensive evaluation of patients for presence of humoral immune response to Ch. trachomatis involves plasma testing of specific antibodies of all three classes. The essential problem in the production of ELISA diagnostic kits is obtaining of positive control. The classic version of positive control is human blood plasma containing specific antibodies. But specific IgM- and IgA-positive sera are deficit raw materials. This fact can significantly limit the production of diagnostic kits, especially in case of large-scale manufacture. We have suggested methodological approach to use of synthetic positive controls in indirect ELISA kits based on conjugate of normal human IgM (IgA and monoclonal antibodies against major outer membrane protein of Ch. trachomatis. It was found that it’s possible to realize such task by means of NHS ester-maleimide-mediated conjugation (by sulfosuccinimidyl-4-(N-maleimidomethylcyclohexane-1-carboxylate and reductive amination-mediated conjugation (by sodium periodate. It was found that synthetic positive controls obtained by different methods are characterized by higher titer compared to IgM- and IgA-positive high

  3. IgM anti-recipient ABO antibodies predict acute graft-versus-host disease following allogeneic hematopoietic stem cell transplantation.

    Science.gov (United States)

    Zaimoku, Yoshitaka; Takami, Akiyoshi; Sato, Hidehiro; Utsumi, Maki; Nakao, Shinji

    2013-07-01

    Passenger lymphocyte syndrome (PLS) presents as transient immune hemolysis due to anti-recipient ABO antibodies produced by donor B-lymphocytes accompanying minor or bidirectional ABO incompatible allogeneic hematopoietic stem cell transplantation (HSCT). We monitored both IgM and IgG type anti-recipient ABO antibodies in 18 consecutive HSCT recipients with hematological malignancies. Five of these patients (28%) developed transient immune hemolysis due to PLS after a median of 19 days post-HSCT. This response was associated with the detection of IgM and IgG anti-recipient ABO antibodies after a median of 16 and 22 days post-HSCT, respectively. All five patients subsequently developed acute graft-versus-host disease (GVHD) grades II-IV, and three died due to transplant-related mortality (TRM) within 1 year after HSCT, while in contrast, of the 13 patients without PLS, three (23%) developed grades II-IV acute GVHD (p ABO antibody may be an early predictor of acute GVHD and poor survival after minor or bidirectional ABO incompatible HSCT.

  4. Seroprevalence of hepatitis B e antigen (HBe antigen and B core antibodies (IgG anti-HBcore and IgM anti-HBcore among hepatitis B surface antigen positive blood donors at a Tertiary Centre in Nigeria

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    Akinbami Akinsegun A

    2012-03-01

    Full Text Available Abstract Background Hepatitis B virus (HBV is a common cause of liver disease throughout the world. HBV is transmitted through blood and other body fluids, including semen and saliva. Chronic replication of HBV virons is characterized by persistence circulation of HBsAg, HBeAg and HBV DNA; usually with anti-HBc and occasionally with anti-HBs. Aim: To determine the prevalence of HBeAg, IgG anti-HBcore and IgM anti-HBcore amongst HBsAg positive blood donors. These parameters are reflective of transmissibility and active hepatitis B infection. A cross sectional study was carried out at the blood donor clinics of Lagos State University Teaching Hospital Ikeja and Lagos University Teaching Hospital Idiaraba. A total of 267 donors were recruited to determine HBe antigen, IgG and IgM anti-HBcore antibodies amongst hepatitis BsAg positive donors. Five milliliters of blood was collected from those who tested positive to HBsAg screen during donation. The sera were subjected to enzyme linked immunosorbent assay (ELISA. Pearson chi-squared test was used for the analytical assessment. Findings A total number of 267 HBsAg positive blood donors were studied. A seroprevalence of 8.2% (22 of 267 HBeAg was obtained, 4 of 267 (1.5% were indeterminate while 241 (90.3% tested negative. Only 27 out of 267 donors (10.1% tested positive to IgM anti-HBcore, 234(87.6% tested negative, while 6(2.2% were indeterminate. A higher percentage of 60.7% (162 of 267 tested positive to IgG anti-HBcore, while 39.3% (105 of 267 tested negative. Conclusion There is a low seroprevalence rate of HBeAg-positive chronic hepatitis and relatively high IgG anti-HBcore and IgM anti-HBcore rates in South West Nigeria.

  5. Guillain-Barré syndrome variant with facial diplegia and paresthesias associated with IgM anti-GalNAc-GD1a antibodies.

    Science.gov (United States)

    Hayashi, Ryuichiro; Yamaguchi, Shigeki

    2015-01-01

    We herein report the case of a 19-year-old woman with facial diplegia and paresthesias (FDP) preceded by flu-like symptoms. We diagnosed the patient with a regional variant of Guillain-Barré syndrome due to decreased tendon reflexes, albuminocytological dissociation in the cerebrospinal fluid and demyelinating features on nerve conduction studies. The patient also had IgM anti-GalNAc-GD1a antibodies, and treatment with glucocorticoids was effective for treating the facial diplegia, but not paresthesia. Therefore, facial palsy may have a different pathophysiology from paresthesia or other symptoms of FDP, which responds to glucocorticoid therapy.

  6. Presence of Mycoplasma fermentans in the bloodstream of Mexican patients with rheumatoid arthritis and IgM and IgG antibodies against whole microorganism

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    Salinas Salvador

    2009-08-01

    Full Text Available Abstract Background Increasing evidence incriminates bacteria, especially Mycoplasma fermentans, as possible arthritogenic agents in humans. The purpose of this study was to investigate M. fermentans in the bloodstream of patients with rheumatoid arthritis. Methods Two hundred and nineteen blood samples from patients with rheumatoid arthritis, systemic lupus erythematosus, antiphospholipid syndrome, and healthy individuals were screened by bacterial culture and direct PCR in order to detect mycoplasmas; IgM and IgG against M. fermentans PG18 were also detected by ELISA and Immunoblotting assays in patients with rheumatoid arthritis and healthy individuals. Results Blood samples from patients with antiphospholipid syndrome and healthy individuals were negative for mycoplasma by culture or direct PCR. In blood samples from patients with systemic lupus erythematosus were detected by direct PCR M. fermentans in 2/50 (2%, M. hominis in 2/50 (2% and U. urealyticum in 1/50 (0.5%. In patients with RA M. fermentans was detected by culture in 13/87 blood samples and in 13/87 by direct PCR, however, there was only concordance between culture and direct PCR in six samples, so M. fermentans was detected in 20/87(23% of the blood samples from patients with RA by either culture or PCR. Antibody-specific ELISA assay to M. fermentans PG18 was done, IgM was detected in sera from 40/87 patients with RA and in sera of 7/67 control individuals, IgG was detected in sera from 48/87 RA patients and in sera from 7/67 healthy individuals. Antibody-specific immunoblotting to M. fermentans PG18 showed IgM in sera from 35/87 patients with RA and in sera from 4/67 healthy individuals, IgG was detected in sera from 34/87 patients and in sera from 5/67 healthy individuals. Conclusion Our findings show that only M. fermentans produce bacteremia in a high percentage of patients with RA. This finding is similar to those reported in the literature. IgM and IgG against M

  7. Detection of IgA and IgM antibodies to HIV-1 in neonates by radioimmune western blotting.

    OpenAIRE

    Portincasa, P.; Conti, G.; Re, M. C.; Chezzi, C.

    1992-01-01

    OBJECTIVE--To detect infection with HIV-1 by IgA and IgM response at birth in children born to HIV-1 seropositive mothers. DESIGN--Western blotting and radioimmune western blotting on stored sera from infected and uninfected babies born to HIV-1 seropositive mothers. Sera were pretreated to remove IgG. SETTING--Parma and Bologna, Italy. SUBJECTS--12 infected and five uninfected babies born to HIV-1 seropositive mothers and three babies born to seronegative mothers. MAIN OUTCOME MEASURES--Effe...

  8. Clinical relevance of different IgG and IgM serum antibody responses to Borrelia burgdorferi after antibiotic therapy for erythema migrans: long-term follow-up study of 113 patients.

    Science.gov (United States)

    Glatz, Martin; Golestani, Marjaneh; Kerl, Helmut; Müllegger, Robert R

    2006-07-01

    To investigate the kinetics of anti-Borrelia burgdorferi antibodies for a minimum of 1 year after antibiotic therapy in patients with erythema migrans (EM) and to correlate antibody titer kinetics with clinical variables. Retrospective study of serial anti-B burgdorferi antibodies in correlation to clinical variables. University-based hospital. One hundred thirteen patients with EM. Pretreatment and a median of 4 consecutive posttreatment serum samples from median follow-up of more than 400 days were simultaneously investigated for anti-B burgdorferi IgG and IgM antibodies. Semiquantitative titers were plotted to identify different groups of antibody kinetics. Individual patients were then stratified to those groups according to their antibody development. A statistical comparison of clinical and therapy-related characteristics among the serologic groups was performed. Anti-B burgdorferi IgG and IgM antibody titers developed in 3 distinct courses: persistent positivity across follow-up (IgG: 12 patients, 11%; IgM: 14, 12%), persistent negativity (IgG: 63, 56%; IgM: 47, 42%), and decrease of a positive pretreatment titer to a negative titer approximately 5 months after therapy (IgG: 34, 30%; IgM: 49, 43%). Statistics revealed significant correlations only between persistent positive IgG titers and long disease duration or large EM lesions before therapy. Long duration or large size of EM before therapy correlates with persistence of a positive anti- B burgdorferi IgG antibody titer after therapy. Serologic profiles do not depend on the type or duration of therapy or the clinical course thereafter. Thus, antibody testing in the follow-up of patients with EM is inappropriate for the assessment of therapeutic response.

  9. Anti-streptococcus IgM antibodies induce repetitive stereotyped movements: cell activation and co-localization with Fcα/μ receptors in the striatum and motor cortex.

    Science.gov (United States)

    Zhang, Danhui; Patel, Ankur; Zhu, Youhua; Siegel, Allan; Zalcman, Steven S

    2012-05-01

    Group A beta-hemolytic streptococcus (GABHS) infections are implicated in neuropsychiatric disorders associated with an increased expression of repetitive stereotyped movements. Anti-streptococcus IgG presumably cross-reacts with elements on basal ganglia cells, modifies their function, and triggers symptoms. IgM may play a unique role in precipitating behavioral disturbances since variations in cortico-striatal activity occur in temporal congruity with peak IgM titers during an orchestrated immune response. We discovered in Balb/c mice that single subcutaneous injections of mouse monoclonal IgM antibodies to streptococcus group A bacteria induce marked dose-dependent increases in repetitive stereotyped movements, including head bobbing, sniffing, and intense grooming. Effects were antibody- and antigen-specific: anti-streptococcus IgG stimulated ambulatory activity and vertical activity but not these stereotypies, while anti-KLH IgM reduced activity. We suggest that anti-streptococcus IgM and IgG play unique roles in provoking GABHS-related behavioral disturbances. Paralleling its stereotypy-inducing effects, anti-streptococcus IgM stimulated Fos-like immunoreactivity in regions linked to cortico-striatal projections involved in motor control, including subregions of the caudate, nucleus accumbens, and motor cortex. This is the first evidence that anti-streptococcus IgM antibodies induce in vivo functional changes in these structures. Moreover, there was a striking similarity in the distributions of anti-streptococcus IgM deposits and Fos-like immunoreactivity in these regions. Of further importance, Fcα/μ receptors, which bind IgM, were present- and co-localized with anti-streptococcus IgM in these structures. We suggest that anti-streptococcus IgM-induced alterations of cell activity reflect local actions of IgM that involve Fcα/μ receptors. These findings support the use of anti-streptococcus monoclonal antibody administration in Balb/c mice to model GABHS

  10. Anti-Streptococcus IgM Antibodies Induce Repetitive Stereotyped Movements: Cell Activation and Co-Localization with Fcα/μ Receptors in the Striatum and Motor Cortex

    Science.gov (United States)

    Zhang, Danhui; Patel, Ankur; Zhu, Youhua; Siegel, Allan; Zalcman, Steven S.

    2012-01-01

    Group A beta-hemolytic streptococcus (GABHS) infections are implicated in neuropsychiatric disorders associated with an increased expression of repetitive stereotyped movements. Anti-streptococcus IgG presumably cross-reacts with elements on basal ganglia cells, modifies their function, and triggers symptoms. IgM may play a unique role in precipitating behavioral disturbances since variations in cortico-striatal activity occur in temporal congruity with peak IgM titers during an orchestrated immune response. We discovered in Balb/c mice that single subcutaneous injections of mouse monoclonal IgM antibodies to Streptococcus Group A bacteria induce marked dose-dependent increases in repetitive stereotyped movements, including head bobbing, sniffing, and intense grooming. Effects were antibody- and antigen-specific: anti-streptococcus IgG stimulated ambulatory activity and vertical activity but not these stereotypies, while anti-KLH IgM reduced activity. We suggest that anti-streptococcus IgM and IgG play unique roles in provoking GABHS-related behavioral disturbances. Paralleling its stereotypy-inducing effects, anti-streptococcus IgM stimulated Fos-like immunoreactivity in regions linked to cortico-striatal projections involved in motor control, including subregions of the caudate, nucleus accumbens, and motor cortex. This is the first evidence that anti-streptococcus IgM antibodies induce in vivo functional changes in these structures. Moreover, there was a striking similarity in the distributions of anti-streptococcus IgM deposits and Fos-like immunoreactivity in these regions. Of further importance, Fcα/μ receptors, which bind IgM, were present- and co-localized with anti-streptococcus IgM in these structures. We suggest that anti-streptococcus IgM-induced alterations of cell activity reflect local actions of IgM that involve Fcα/μ receptors. These findings support the use of anti-streptococcus monoclonal antibody administration in Balb/c mice to model GABHS

  11. Immunization with gingipain A hemagglutinin domain of Porphyromonas gingivalis induces IgM antibodies binding to malondialdehyde-acetaldehyde modified low-density lipoprotein.

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    Mikael Kyrklund

    Full Text Available Treatment of periodontitis has beneficial effects on systemic inflammation markers that relate to progression of atherosclerosis. We aimed to investigate whether immunization with A hemagglutinin domain (Rgp44 of Porphyromonas gingivalis (Pg, a major etiologic agent of periodontitis, would lead to an antibody response cross-reacting with oxidized low-density lipoprotein (OxLDL and how it would affect the progression of atherosclerosis in low-density lipoprotein receptor-deficient (LDLR-/- mice. The data revealed a prominent IgM but not IgG response to malondialdehyde-acetaldehyde modified LDL (MAA-LDL after Rgp44 and Pg immunizations, implying that Rgp44/Pg and MAA adducts may share cross-reactive epitopes that prompt IgM antibody production and consequently confer atheroprotection. A significant negative association was observed between atherosclerotic lesion and plasma IgA to Rgp44 in Rgp44 immunized mice, supporting further the anti-atherogenic effect of Rgp44 immunization. Plasma IgA levels to Rgp44 and to Pg in both Rgp44- and Pg-immunized mice were significantly higher than those in saline control, suggesting that IgA to Rgp44 could be a surrogate marker of immunization in Pg-immunized mice. Distinct antibody responses in plasma IgA levels to MAA-LDL, to Pg lipopolysaccharides (Pg-LPS, and to phosphocholine (PCho were observed after Rgp44 and Pg immunizations, indicating that different immunogenic components between Rpg44 and Pg may behave differently in regard of their roles in the development of atherosclerosis. Immunization with Rgp44 also displayed atheroprotective features in modulation of plaque size through association with plasma levels of IL-1α whereas whole Pg bacteria achieved through regulation of anti-inflammatory cytokine levels of IL-5 and IL-10. The present study may contribute to refining therapeutic approaches aiming to modulate immune responses and inflammatory/anti-inflammatory processes in atherosclerosis.

  12. UltramicroELISA para la detección de anticuerpos IgM anti M. leprae UltramicroELISA assay for the detection of human IgM antibodies to M. leprae

    Directory of Open Access Journals (Sweden)

    José Laferte

    1991-12-01

    Full Text Available La disponibilidad del sistema Ultramicroanalítico (SUMA y de un antígeno especie-específico del M. leprae obtenido mediante síntesis química, permitió la normalización y validación de un ultramicroELISA para la detección de anticuerpos IgM específicos a esta micobacteria. El análisis de 433 sueros de banco de sangre y 265 sueros usados para validar el método y clasificados en un grupo control de donantes de banco de sangre (100, un grupo de pacientes tuberculosos (50, un grupo de enfermos de lepra (65 y un grupo de contactos de estos enfermos (50, mostró la especificidad del ensayo para evidenciar la infección con el M. leprae. Los resultados obtenidos del estudio adicional de 140 muestras de suero de contactos de enfermos estuvieron estrechamente correlacionados (r = 0,98 con los resultados obtenidos por la técnica de microELISA convencional. La utilización del SUMA no solo permite un notable ahorro de reactivos si no además facilita la lectura, cálculo, validación y almacenamiento automático de los resultados.The availability of an ultramicroanalitic system (SUMA and specie-specific antigen of M. leprae obtained by chemical synthesis, have made possible the standardization and validation of an ultramicroELISA assay for detecting specific human IgM antibodies to this mycobacterium. The specificity of this test to demonstrate the infection with M. leprae was corroborated through a screening of 433 blood bank serum samples and other 265 from diferent groups (100, control group, 50 tuberculosis patients, 65 leprosy patients, 50 from household. The results obtained in the aditional study of 140 household sero showed a high correlation (r = 0.98 with the conventional microELISA method. The use of SUMA allows saving reagents and time since sample handling, plate reading, print out and storing the data are computer assisted.

  13. The natural human IgM antibody PAT-SM6 induces apoptosis in primary human multiple myeloma cells by targeting heat shock protein GRP78.

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    Leo Rasche

    Full Text Available In contrast to other haematological malignancies, targeted immunotherapy has not entered standard treatment regimens for de novo or relapsed multiple myeloma (MM yet. While a number of IgG-formatted monoclonal antibodies are currently being evaluated in clinical trials in MM, our study aimed to investigate whether the fully human IgM monoclonal antibody PAT-SM6 that targets a tumour-specific variant of the heat shock protein GRP78 might be an attractive candidate for future immunotherapeutic approaches. We here show that GRP78 is stably and consistently expressed on the surface on tumour cells from patients with de novo, but also relapsed MM and that binding of PAT-SM6 to MM cells can specifically exert cytotoxic effects on malignant plasma cells, whereas non-malignant cells are not targeted. We demonstrate that the induction of apoptosis and, to a lesser extent, complement dependent cytotoxicity is the main mode of action of PAT-SM6, whereas antibody dependent cellular cytotoxicity does not appear to contribute to the cytotoxic properties of this antibody. Given the favourable safety profile of PAT-SM6 in monkeys, but also in a recent phase I trial in patients with malignant melanoma, our results form the basis for a planned phase I study in patients with relapsed MM.

  14. Anti-Fas2 IgM antibodies in Fasciola hepatica infected patients with positive IgG serology.

    Science.gov (United States)

    Kazantseva, Liliya; García Lázaro, María Del Pilar; Herrera-Velit, Patricia; Espinoza, Jose R

    2017-03-01

    Fascioliasis is an infectious disease caused by parasites Fasciola hepatica and F. gigantica. Humans are infected by the consumption of vegetables and water contaminated with the infective form of the parasite. In this study, an IgM-ELISA with the cysteine proteinase Fas2 antigen was evaluated with sera from 76 patients infected with F. hepatica, 24 patients with other parasite infections and 84 healthy volunteers. IgM-ELISA resulted in 43% positives in F. hepatica patients with positive serology to Fas2-ELISA, but no positives resulted from testing healthy volunteers and individuals infected with other parasites. The IgM-ELISA diagnostic parameters showed a sensitivity of 43.4% (95% CI 0.321-0.553), specificity of 100% (95% CI 0.957-1), and no cross-reactivity with other parasitic infection. Interference by rheumatoid factor in the IgM immunoassay was controlled by treating sera with rheumatoid factor absorbent before testing. Fas2 antigen is detected by circulating IgM in patients infected with F. hepatica and IgM-ELISA using Fas2 appears as a specific immunoassay to detect the acute phase of the acute phase of F. hepatica infection in humans. © The Author 2017. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  15. Fully human IgG and IgM antibodies directed against the carcinoembryonic antigen (CEA Gold 4 epitope and designed for radioimmunotherapy (RIT of colorectal cancers

    Directory of Open Access Journals (Sweden)

    Pugnière Martine

    2004-10-01

    Full Text Available Abstract Background Human monoclonal antibodies (MAbs are needed for colon cancer radioimmunotherapy (RIT to allow for repeated injections. Carcinoembryonic antigen (CEA being the reference antigen for immunotargeting of these tumors, we developed human anti-CEA MAbs. Methods XenoMouse®-G2 animals were immunized with CEA. Among all the antibodies produced, two of them, VG-IgG2κ and VG-IgM, were selected for characterization in vitro in comparison with the human-mouse chimeric anti-CEA MAb X4 using flow cytometry, surface plasmon resonance, and binding to radiolabeled soluble CEA and in vivo in human colon carcinoma LS174T bearing nude mice. Results Flow cytometry analysis demonstrated binding of MAbs on CEA-expressing cells without any binding on NCA-expressing human granulocytes. In a competitive binding assay using five reference MAbs, directed against the five Gold CEA epitopes, VG-IgG2κ and VG-IgM were shown to be directed against the Gold 4 epitope. The affinities of purified VG-IgG2κ and VG-IgM were determined to be 0.19 ± 0.06 × 108 M-1 and 1.30 ± 0.06 × 108 M-1, respectively, as compared with 0.61 ± 0.05 × 108 M-1 for the reference MAb X4. In a soluble phase assay, the binding capacities of VG-IgG2κ and VG-IgM to soluble CEA were clearly lower than that of the control chimeric MAb X4. A human MAb concentration of about 10-7 M was needed to precipitate approximatively 1 ng 125I-rhCEA as compared with 10-9 M for MAb X4, suggesting a preferential binding of the human MAbs to solid phase CEA. In vivo, 24 h post-injection, 125I-VG-IgG2κ demonstrated a high tumor uptake (25.4 ± 7.3%ID/g, close to that of 131I-X4 (21.7 ± 7.2%ID/g. At 72 h post-injection, 125I-VG-IgG2κ was still concentrated in the tumor (28.4 ± 11.0%ID/g whereas the tumor concentration of 131I-X4 was significantly reduced (12.5 ± 4.8%ID/g. At no time after injection was there any accumulation of the radiolabeled MAbs in normal tissues. A pertinent analysis of

  16. User's guide to the indirect solid-phase radioimmunoassay for the detection of cytomegalovirus-specific IgM antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Griffiths, P.D. (Royal Free Hospital School of Medicine, London (UK). Virus Laboratory); Kangro, H.O. (St. Bartholomew' s Hospital, London (UK). Virology Dept.)

    1984-07-01

    In this review full laboratory details are given of a solid-phase indirect radioimmunoassay for the detection of specific IgM antibodies against cytomegalovirus. Practical advice is given on readily available commercial sources of reagents, a simple iodination procedure, the rapid dilution of sera under test and calculation of the results with a computer program available from the authors. Problems encountered with the assay are also detailed such as interference by rheumatoid factor, deterioration of the radiolabel and high background binding found with some sera. If these problems are avoided by the methods given in the text then the radioimmunoassay has been shown to give results of 100% specificity with 89% sensitivity for detecting congenital infection and 92% sensitivity for identifying primary cytomegalovirus infection in pregnant women.

  17. A user's guide to the indirect solid-phase radioimmunoassay for the detection of cytomegalovirus-specific IgM antibodies

    International Nuclear Information System (INIS)

    Griffiths, P.D.; Kangro, H.O.

    1984-01-01

    In this review full laboratory details are given of a solid-phase indirect radioimmunoassay for the detection of specific IgM antibodies against cytomegalovirus. Practical advice is given on readily available commercial sources of reagents, a simple iodination procedure, the rapid dilution of sera under test and calculation of the results with a computer program available from the authors. Problems encountered with the assay are also detailed such as interference by rheumatoid factor, deterioration of the radiolabel and high background binding found with some sera. If these problems are avoided by the methods given in the text then the radioimmunoassay has been shown to give results of 100% specificity with 89% sensitivity for detecting congenital infection and 92% sensitivity for identifying primary cytomegalovirus infection in pregnant women. (Auth.)

  18. Immunodominant IgM and IgG Epitopes Recognized by Antibodies Induced in Enterovirus A71-Associated Hand, Foot and Mouth Disease Patients.

    Directory of Open Access Journals (Sweden)

    Kam Leng Aw-Yong

    Full Text Available Enterovirus A71 (EV-A71 is one of the main causative agents of hand, foot and mouth disease (HFMD. Unlike other enteroviruses that cause HFMD, EV-A71 is more frequently associated with severe neurological complications and fatality. To date, no effective licensed antivirals are available to combat EV-A71 infection. Little is known about the immunogenicity of viral non-structural proteins in humans. Previous studies have mainly focused on characterization of epitopes of EV-A71 structural proteins by using immunized animal antisera. In this study, we have characterized human antibody responses against the structural and non-structural proteins of EV-A71. Each viral protein was cloned and expressed in either bacterial or mammalian systems, and tested with antisera by western blot. Results revealed that all structural proteins (VP1-4, and non-structural proteins 2A, 3C and 3D were targets of EV-A71 IgM, whereas EV-A71 IgG recognized all the structural and non-structural proteins. Sixty three synthetic peptides predicted to be immunogenic in silico were synthesized and used for the characterization of EV-A71 linear B-cell epitopes. In total, we identified 22 IgM and four IgG dominant epitopes. Synthetic peptide PEP27, corresponding to residues 142-156 of VP1, was identified as the EV-A71 IgM-specific immunodominant epitope. PEP23, mapped to VP1 41-55, was recognized as the EV-A71 IgG cross-reactive immunodominant epitope. The structural protein VP1 is the major immunodominant site targeted by anti-EV-A71 IgM and IgG antibodies, but epitopes against non-structural proteins were also detected. These data provide new understanding of the immune response to EV-A71 infection, which benefits the development of diagnostic tools, potential therapeutics and subunit vaccine candidates.

  19. Radioimmunological determination of growth hormone antibodies

    International Nuclear Information System (INIS)

    Kracmar, P.; Hnikova, O.

    1979-01-01

    The method is based on the assumption of the presence of antibodies in the serum of the patient and the formation of the complex antibody-tracer ( 125 I-STH). For separation the principle is used of two antibodies and subsequent ultrafiltration with membrane ultrafilters. Clinical experience, reproducibility and the procedure recommended for simple monitoring and the determination of the amount of antibodies in the serum of patients are presented. (author)

  20. Tumour localization and pharmacokinetics of iodine-125 human monoclonal IgM antibody (COU-1) and its monomeric and half-monomeric fragments analysed in nude mice grafted with human tumour

    International Nuclear Information System (INIS)

    Ditzel, H.; Erb, K.; Rasmussen, J.W.; Jensenius, J.C.

    1992-01-01

    Human monoclonal IgM antibodies reactive with cancer-associated antigens may not have the optimal imaging capability due to their large size. Fragmentation of human IgM is less than straight-forward due to the loss of immunoreactivity. From the human monoclonal IgM antibody COU-1 we have prepared monomeric and half-monomeric fragments, which retain the ability to bind to colon cancer cells in vitro. The pharmacokinetics and tumour localization were evaluated in nude mice bearing human colon adenocarcinoma and human melanoma grafts. Faster clearance from the circulation was seen for the smaller half-monomeric fragment with a half-life (rapid phase/slow phase) of 2 h/16 h compared with the intact antibody, 4 h/25 h, and the monomeric fragment, 3 h/27 h. Intact COU-1 as well as the fragments accumulated in the colon tumour graft. Higher amounts of radioactivity were found in the colon tumour as compared to normal organs for intact COU-1 at days 4 and 6, for the monomeric fragment at day 4, and for the half-monomeric fragment at day 2 after injection. This investigation demonstrates the favourable biodistribution of the half monomeric COU-1 fragment. The fast clearance of this fragment resulted in a tumour-to-muscle ratio as high as 22 on day 2 after injection. Also, only this fragment gave a positive tumour-to-blood ratio. Normal IgM and its fragments were used as controls. Radioimmunoscintigraphy demonstrated the colon tumour discriminatory properties of each of the three iodine-labelled antibody preparations. The results compare favourably with previously reported investigations of the localization of human monoclonal antibodies and suggest that fragments of human monoclonal IgM antibodies may be useful tools for the immunodetection of cancer in patients. (orig.)

  1. C-reactive protein and natural IgM antibodies are activators of complement in a rat model of intestinal ischemia and reperfusion

    NARCIS (Netherlands)

    Padilla, Niubel Diaz; van Vliet, Arlene K.; Schoots, Ivo G.; Seron, Mercedes Valls; Maas, M. Adrie; Peltenburg, Esther E. Posno; de Vries, Annebeth; Niessen, Hans W. M.; Hack, C. Erik; van Gulik, Thomas M.

    2007-01-01

    Background. The role of C-reactive protein (CRP), natural immunoglobulin M (IgM), and natural IgM against phosphorylcholine (anti-Pc IgM) was investigated in relation with complement activation in a rat model of intestinal ischemia and reperfusion (II/R). The effect of Cl-esterase inhibitor (C1-Inh)

  2. Altered antibody response to influenza H1N1 vaccine in healthy elderly people as determined by HI, ELISA, and neutralization assay

    NARCIS (Netherlands)

    Remarque, E.J.; Bruijn, de I.A.; Boersma, W.J.A.; Masurel, N.; Ligthart, G.J.

    1998-01-01

    To determine the influence of ageing per se as well as of priming histories on the antibody response to influenza vaccination, haemagglutination inhibition (HI), ELISA IgG, IgA, IgM and neutralizing antibody titres were studied in 43 healthy young subjects (mean age 23 years) and 55 healthy elderly

  3. A high-affinity human monoclonal IgM antibody reacting with multiple strains of Mycoplasma hominis

    DEFF Research Database (Denmark)

    Moller, SA; Birkelund, Svend; Borrebaeck, CA

    1990-01-01

    Human monoclonal antibodies were produced against Mycoplasma hominis by in vitro immunization of peripheral blood lymphocytes from a healthy seropositive donor using low amounts of antigen (5 ng/ml). The immune B lymphocytes were subsequently immortalized by Epstein-Barr virus transformation foll...

  4. A carrier for non-covalent delivery of functional beta-galactosidase and antibodies against amyloid plaques and IgM to the brain.

    Directory of Open Access Journals (Sweden)

    Gobinda Sarkar

    Full Text Available BACKGROUND: Therapeutic intervention of numerous brain-associated disorders currently remains unrealized due to serious limitations imposed by the blood-brain-barrier (BBB. The BBB generally allows transport of small molecules, typically <600 daltons with high octanol/water partition coefficients, but denies passage to most larger molecules. However, some receptors present on the BBB allow passage of cognate proteins to the brain. Utilizing such receptor-ligand systems, several investigators have developed methods for delivering proteins to the brain, a critical requirement of which involves covalent linking of the target protein to a carrier entity. Such covalent modifications involve extensive preparative and post-preparative chemistry that poses daunting limitations in the context of delivery to any organ. Here, we report creation of a 36-amino acid peptide transporter, which can transport a protein to the brain after routine intravenous injection of the transporter-protein mixture. No covalent linkage of the protein with the transporter is necessary. APPROACH: A peptide transporter comprising sixteen lysine residues and 20 amino acids corresponding to the LDLR-binding domain of apolipoprotein E (ApoE was synthesized. Transport of beta-galactosidase, IgG, IgM, and antibodies against amyloid plques to the brain upon iv injection of the protein-transporter mixture was evaluated through staining for enzyme activity or micro single photon emission tomography (micro-SPECT or immunostaining. Effect of the transporter on the integrity of the BBB was also investigated. PRINCIPAL FINDINGS: The transporter enabled delivery to the mouse brain of functional beta-galactosidase, human IgG and IgM, and two antibodies that labeled brain-associated amyloid beta plaques in a mouse model of Alzheimer's disease. SIGNIFICANCE: The results suggest the transporter is able to transport most or all proteins to the brain without the need for chemically linking the

  5. Validation of an ELISA for the concurrent detection of total antibodies (IgM and IgG to Rift Valley fever virus

    Directory of Open Access Journals (Sweden)

    Charlotte E. Ellis

    2014-05-01

    Full Text Available Rift Valley fever virus (RVFV infects humans and livestock, causing haemorrhaging andabortions in animals. Three major RVF epizootics have occurred in South Africa since the1950s and the outbreak in 2010 had a mortality rate of 10.7% in humans. Accurate and earlydetection is therefore essential for management of this zoonotic disease. Enzyme-linkedimmunosorbent assays (ELISAs have been developed for the detection of either IgM or IgGantibodies to RVFV in animal sera. In this study, data are presented on the validation of adouble-antigen ELISA for the simultaneous detection of both classes of antibodies to RVFV ina single test. ELISA plates were coated with a recombinant nucleoprotein. The nucleoprotein,conjugated to horseradish peroxidase, was used as the detecting reagent. A total of 534 serafrom sheep and cattle were used in the validation. The sheep sera were collected during a RVFpathogenesis study at the Agricultural Research Council (ARC – Onderstepoort VeterinaryInstitute and the cattle sera were collected during an outbreak of RVF in 2008 at the ARC –Animal Production Institute in Irene, Pretoria. The ELISA had a diagnostic sensitivity of 98.4%and a specificity of 100% when compared to a commercial cELISA. This convenient and fastassay is suitable for use in serological surveys or monitoring immune responses in vaccinatedanimals.

  6. Multiplex assay (Mikrogen recomBead) for detection of serum IgG and IgM antibodies to 13 recombinant antigens of Borrelia burgdorferi sensu lato in patients with neuroborreliosis

    DEFF Research Database (Denmark)

    Dessau, Ram Benny; Møller, Jens K.; Kolmos, Birte

    2015-01-01

    A multiplex-bead-based assay for the detection of serum antibodies to Borrelia burgdorferi sensu lato was evaluated. The assay contained 13 different antigens in both the IgG and the IgM assay; thus, a total of 26 measurement results were available from each sample. A total of 49 Danish patients......%, respectively. Overall, the patients with LNB had serum reactivity in IgG VlsE, but modest antibody reactivity in the remaining 12 IgG and 13 IgM antibody measurements. Using a logistic regression model with five IgG and two IgM antigens, the sensitivity and specificity of the assay was improved; but the Ig...... with Lyme neuroborreliosis (LNB), 218 Danish blood donor controls, a set of 61 Swedish patients with LNB and 139 Swedish non-LNB patients investigated for suspected LNB were used. There are four parts developed in this study: a characterization of the sero-epidemiological antibody-response pattern...

  7. Towards Universal Screening for Toxoplasmosis: Rapid, Cost-effective and Simultaneous Detection of Toxoplasma Anti-IgG, IgM and IgA Antibodies Using Very Small Serum Volumes

    Science.gov (United States)

    No dataset associated with this publication.This dataset is associated with the following publication:Augustine, S. Towards Universal Screening for Toxoplasmosis: Rapid, Cost-effective and Simultaneous Detection of Toxoplasma Anti-IgG, IgM and IgA Antibodies Using Very Small Serum Volumes. JOURNAL OF CLINICAL MICROBIOLOGY. American Society for Microbiology, Washington, DC, USA, 56(7): 1-2, (2016).

  8. Utilização, em politransfundidos, da pesquisa de anticorpos igm anti-trypanosoma cruzi e anti-toxoplasma gondii para detectar infecções pós-transfusionais recentes IgM Trypanosoma cruzi and Toxoplasma gondii antibodies in the detection of recent transfusion-transmitted infections

    Directory of Open Access Journals (Sweden)

    Vicente Amato Neto

    1984-04-01

    Full Text Available Consideram os Autores que a pesquisa de anticorpos IgM no soro é tática capaz de revelar recentes infecções pós-transfusionais. Por isso, decidiram usar esse tipo de mensuração relativamente a grupo constituído por 101 politrans-fundidos, tendo abordado especificamente as aquisições de doença de Chagas e toxoplasmose. Através da investigação que realizaram, só em duas oportunidades encontraram anticorpos IgM anti-Trypanosoma cruzi ou anti-Toxoplasma gondii e, portanto, não evidenciaram expressivo panorama tradutor de processos há pouco tempo contraídos, como ainda, por meio de anticorpos IgG não identificaram números expressivos de pessoas com essas protozooses. No entanto, detectaram a expressiva taxa de 4,9% de casos de doença de Chagas muito provavelmente decorrentes da hemoterapia. A despeito da relevância não acentuada dos resultados que obtiveram, julgaram os Autores ser válido estimular a efetivação de outros estudos congêneres e correlatos, aptos a contribuir para aqui-latamento de riscos pertinentes à prática hemoterápica.The Authors have regarded serum IgM antibodies titration as useful in the detection of recent transfusion-transmitted infections. For this reason a group consisting of 101 patients, who had received many blood transfusions, underwent such mensuration in order to reveal recent Chagas'disease and toxoplasmosis acquired infections. Throughout the investigation just two cases have yielded IgM trypanosomal or toxoplasmal antibodies, showing therefore that this sort of titration did not correlate with the real existence of recent acquired infections. On the other hand IgM antibodies in the same patients did not show a considerable incidence of these two protozoan infections. However an expressive rate of 4.9% of Chagas'disease probably due to hemotherapy was found. Although the results this study were not very relevant, the Authors still have in mind that further similar investigations should be

  9. Enzyme-linked immunoassay for dengue virus IgM and IgG antibodies in serum and filter paper blood

    NARCIS (Netherlands)

    Tran, Thanh Nga T.; de Vries, Peter J.; Hoang, Lan Phuong; Phan, Giao T.; Le, Hung Q.; Tran, Binh Q.; Vo, Chi Mai T.; Nguyen, Nam V.; Kager, Piet A.; Nagelkerke, Nico; Groen, Jan

    2006-01-01

    BACKGROUND: The reproducibilty of dengue IgM and IgG ELISA was studied in serum and filter paper blood spots from Vietnamese febrile patients. METHODS: 781 pairs of acute (t0) and convalescent sera, obtained after three weeks (t3) and 161 corresponding pairs of filter paper blood spots were tested

  10. Enzyme-linked immunoassay for dengue virus IgM and IgG antibodies in serum and filter paper blood

    NARCIS (Netherlands)

    Tran, T.N.T.; de Vries, P.J.; Hoang, L.P.; Phan, G.T.; Le, H.Q.; Tran, B.Q.; Vo, C.M.T.; Nguyen, N.V.; Kager, P.A.; Nagelkerke, N.; Groen, J.

    2006-01-01

    Background: The reproducibilty of dengue IgM and IgG ELISA was studied in serum and filter paper blood spots from Vietnamese febrile patients. Methods: 781 pairs of acute (t0) and convalescent sera, obtained after three weeks (t3) and 161 corresponding pairs of filter paper blood spots were tested

  11. Two years' performance of an in-house ELISA for diagnosis of Legionnaires' disease: detection of specific IgM and IgG antibodies against Legionella pneumophila serogroup 1, 3 and 6 in human serum.

    Science.gov (United States)

    Elverdal, P L; Jørgensen, C S; Krogfelt, K A; Uldum, S A

    2013-08-01

    The aim of this study was to evaluate the performance of an in-house ELISA for the diagnosis of Legionnaires' disease (LD) by detection of IgM and IgG antibodies against Legionella (L.) pneumophila serogroups (sg) 1, 3 and 6. The evaluation was done throughout a two-year period in a diagnostic routine laboratory. Furthermore, the sensitivity of four different methods, the in-house L. pneumophila antibody test (ELISA), the urinary antigen test (Binax® EIA), an in-house PCR and culture, both alone and in combination was evaluated. From 2008 to 2010, 12,158 serum samples from 10,503 patients were analysed. During the same period, 361 cases of laboratory-confirmed LD cases were recorded in Denmark, but of these only 113 had a serum sample examined. The positive predictive value of the in-house ELISA was calculated to be 12.8 and the negative predictive value was 99.6, using only the confirmed LD cases as true positives. The sensitivity of the in-house ELISA for the detection of IgM and IgG antibodies in the confirmed LD cases was 61% and 36%, respectively. By combining the two ELISA assays the sensitivity increased to 66%. The sensitivity of the Legionella urinary antigen test (Binax® EIA) was 63%, of the in-house PCR 87% and of culture 69%. When all the different methods were combined, a higher sensitivity was calculated--for in-house ELISA (IgM+IgG) and Binax® EIA 91%, in-house ELISA (IgM+IgG) and in-house PCR 93%, in-house ELISA (IgM+IgG) and culture 93%, Binax® EIA and in-house PCR 79%, Binax® EIA and culture 68% and in-house PCR and culture 94%. This study confirms that the detection of IgG and IgM antibodies by ELISA is an important diagnostic tool, also during the initial phase of the disease. Furthermore, we showed that LD in Denmark with or without serum samples collected exhibits the same age and sex distribution and epidemiology, as in the rest of Europe, i.e., mostly men are infected, infections are mostly community acquired, followed by infection from

  12. Beta1 integrin is not essential for hematopoiesis but is necessary for the T cell-dependent IgM antibody response

    DEFF Research Database (Denmark)

    Brakebusch, Cord; Fillatreau, Simon; Potocnik, Alexandre J

    2002-01-01

    Several experimental evidences suggested that beta1 integrin-mediated adhesion of hematopoietic stem cells (HSC) is important for their function in the bone marrow (BM). Using induced deletion of the beta1 integrin gene restricted to the hematopoietic system, we show that beta1 integrin...... is not essential for HSC retention in the BM, hematopoiesis, and trafficking of lymphocytes. However, immunization with a T cell-dependent antigen resulted in virtually no IgM production and an increased secretion of IgG in mutant mice, while the response to a T cell-independent type 2 antigen showed decreases...

  13. IGMS Construction Grants Overview

    Science.gov (United States)

    The Integrated Grants Management System (IGMS) is a web-based system that contains information on the recipient of the grant, fellowship, cooperative agreement and interagency agreement, including the name of the entity accepting the award.

  14. Determination of antiphospholipid antibodies and Thrombophilia in ...

    African Journals Online (AJOL)

    Background: Recurrent miscarriage is a critical problem in which many factors play a crucial role such as antiphospholipid antibodies (APA) and anticardiolipin antibodies (ACA). Recent studies pointed to a potential role of thrombophilias as a possible cause of recurrent miscarriage (RM). Objectives: This study was ...

  15. Development of a novel protein biochip enabling validation of immunological assays and detection of serum IgG and IgM antibodies against Treponema pallidum pathogens in the patients with syphilis.

    Science.gov (United States)

    Huang, Na-Li; Ye, Lei; Schneider, Marion E; Du, Yi-Xin; Xu, Yuan-Hong; Fan, Li-Bin; Du, Wei-Dong

    2016-01-15

    In this study, we developed a novel protein biochip methodology that was characterized by dithiobis (succinimidyl undecanoate) (DSU) and specialized for detection of serum IgG and IgM antibodies against Treponema pallidum pathogens in the patients with syphilis, respectively. The biochips were validated by a dimension of atomic force microscope (AFM). The visualized detection limit of IgG antibody on the biochip was 0.39μg/ml. Finally, 286 serum samples from the patients with syphilis were simultaneously tested on the rTpN15-17-47 coated biochips. The results were evaluated in comparison with the assays of T. pallidum particle agglutination (TPPA) and the toluidine red unheated serum test (TRUST). The result demonstrated that the relative positive rate in the 286 patients by biochip was 99.0%, similar to that by TPPA (97.9%, P>0.05) and higher than that by TRUST, (76.2%, Ppallidum on a larger scale, but also for monitoring therapy modality of the disease. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Development of ELISAs for quantification of HMFG1-specific human anti-mouse IgG and IgM antibodies.

    Science.gov (United States)

    Oei, A L M; Boerman, O C; Geurts-Moespot, A; van Eerd, J E; van Tienoven, D; Courtenay-Luck, N; Thomas, C M G; Massuger, L F A G; Sweep, F C G J

    2007-01-01

    The aim of this study was to develop and validate ELISAs for quantification of HAMA-IgM and HAMA-IgG in serum of patients with ovarian cancer who enrolled in a large international randomized phase III trial of intraperitoneal Yttrium-90-labeled HMFG1 murine monoclonal antibody therapy. The capture antibody of these 2 assays was the murine antibody HMFG1, while mouse anti-human IgM-HRP or mouse anti-human IgG(Fc)-HRP served as tracer antibodies. A pool of HAMA-positive serum samples was used to prepare a series of assay standards and another pool served as reference preparation. The analytical sensitivity of the HAMA-IgM assay was 2.5 arbitrary units per mL (AU/mL) and 4.7 AU/mL for the HAMA-IgG ELISA. Diluted serum samples showed good parallelism with the HAMA-IgM and HAMA-IgG standard dose-response curves. Within-assay coefficient of variation was 7.5% for HAMA-IgM and 6.5% for HAMA-IgG. Between-assay variation was 14.2% for HAMA-IgM and 15.3% for HAMA-IgG. The developed HAMA-IgM and HAMA-IgG ELISAs show satisfactory reliability criteria (sensitivity, parallelism and precision) and are suitable for monitoring of HAMA-IgM and HAMA-IgG responses in ovarian cancer patients. These ELISAs will be used to monitor the development of HAMAs in patients who received radioimmunotherapy with murine HMFG1.

  17. Do monoclonal antibodies recognize linear sequential determinants?

    Science.gov (United States)

    Camera, M; Muratti, E; Trinca, M L; Chersi, A

    1988-01-01

    A group of 19 anti-class II monoclonal antibodies produced in different laboratories were tested in ELISA for their ability to bind to a panel of synthetic peptides selected from HLA-DQ alpha and beta chains. No one of the antibodies tested was found to react with the synthetic fragments, thus confirming the common finding that MoAbs generally fail to recognize fragments of the native antigen. The possibility that this result might be partly due to the procedure used for screening hybridoma supernatants is discussed.

  18. Anticuerpos antinucleares, imágenes y características obtenidas por inmunofluorescencia: Importancia de los isotipos IgA, IgM e IgG Antinuclear antibodies, patterns and characteristics obtained by immunofluorescence: The importance of the IgA, IgM and IgG isotypes

    Directory of Open Access Journals (Sweden)

    Miriam Arcavi

    2009-10-01

    Full Text Available La técnica de elección para el screening de anticuerpos antinucleares (ANA es la inmunofluorescencia indirecta que utiliza como sustrato una línea de células epiteliales de carcinoma de laringe humano (IFI-HEp2, y como antisuero, anti-IgG o anti-Ig totales. Los ANA-IgG son los más importantes para el diagnóstico y monitoreo de las enfermedades del tejido conectivo (ETC, mientras los ANA-IgM son de menor relevancia clínica en estos pacientes. Sin embargo, poco se sabe de los ANA-IgA ya que estos Ac han sido menos investigados. El objetivo de este trabajo fue estudiar la prevalencia de los diferentes isotipos de inmunoglobulinas de anticuerpos antinucleares en los pacientes con ETC y evaluar la conveniencia de utilizar conjugados monovalentes o polivalentes. Se procesaron 100 sueros de pacientes con diversas ETC empleando IFI-HEp2, en los cuales se detectó 38% de ANA-IgA (títulos ≥ 1:80 y 12% de ANA-IgM (títulos ≤ 1:160. En 29 casos se detectó IgA en ausencia de IgM, en 3 casos IgM en ausencia de IgA. En todos los casos los ANA-IgG estuvieron presentes. En 6 sueros se observó un cambio de imagen con conjugado anti-IgA y en 3 con conjugado anti-IgM. Debido a la alta prevalencia de ANA-IgA detectada por IFI-HEp2, se destaca la conveniencia de utilizar conjugado anti-Ig totales en lugar de anti-IgG, mientras se desconozca la relevancia de los ANA-IgA en el diagnóstico, pronóstico y seguimiento de las enfermedades reumáticas sistémicas.The indirect immunofluorescence with epitelial cell line from human laryngeal carcinoma as substrate (IIF-HEp2 and anti-IgG or anti-total Ig as antisera, is the technique currently used for the detection of antinuclear antibodies. The most important antibodies for the diagnosis and follow-up of connective tissue diseases (CTD are the IgG-ANA, while the IgM-ANA have no clinical relevance. However the IgA-ANA have not been thoroughly investigated so far. The aim of this work was to study the prevalence

  19. IgM repertoire biodiversity is reduced in HIV-1 infection and systemic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    Li eYin

    2013-11-01

    Full Text Available Background: HIV-1 infection or systemic lupus erythematosus [SLE] disrupt B cell homeostasis, reduce memory B cells, and impair function of IgG and IgM antibodies. Objective: To determine how disturbances in B cell populations producing polyclonal antibodies relate to the IgM repertoire, the IgM transcriptome in health and disease was explored at the complementarity determining region 3 [CDRH3] sequence population level. Methods: 454-deep pyrosequencing in combination with a novel analysis pipeline was applied to define populations of IGHM CDRH3 sequences based on absence or presence of somatic hypermutations [SHM] in peripheral blood B cells. Results: HIV or SLE subjects have reduced biodiversity within their IGHM transcriptome compared to healthy subjects, mainly due to a significant decrease in the number of unique combinations of alleles, although recombination machinery was intact. While major differences between sequences without or with SHM occurred among all groups, IGHD and IGHJ allele use, CDRH3 length distribution, or generation of SHM were similar among study cohorts. Antiretroviral therapy failed to normalize IGHM biodiversity in HIV-infected individuals. All subjects had a low frequency of allelic combinations within the IGHM repertoire similar to known broadly-neutralizing HIV-1 antibodies. Conclusions: Polyclonal expansion would decrease overall IgM biodiversity independent of other mechanisms for development of the B cell repertoire. Applying deep sequencing as a strategy to follow development of the IgM repertoire in health and disease provides a novel molecular assessment of multiple points along the B cell differentiation pathway that is highly sensitive for detecting perturbations within the repertoire at the population level.

  20. Simple hanging drop (20 μl) double antibody radioimmunoassay of human IgM, IgG, IgA, IgD and IgE

    International Nuclear Information System (INIS)

    Newton, C.A.; Pereira, R.S.; Farrant, J.

    1981-01-01

    A simple hanging drop radioimmunoassay has been developed for different classes of human immunoglobulin from serum or culture supernatant samples. The immune precipitate was formed in inverted Terasaki plates (20 μl per well) using 4 aliquots (5 μl) as follows: (A) a standard or unknown sample; (B) rabbit antisera to the appropriate immunoglobulin class; (C) 125 I-labelled human immunoglobulin of the appropriate class; (D) a developing goat-anti rabbit serum. The immune precipitate in the hanging drop was harvested and washed without centrifugation with a simple Terasaki plate harvester. The method gives a great saving both in time and in sample and reagent volumes in comparison with an established double antibody assay in tubes (4 x 100 μl aliquots). (Auth.)

  1. Enzyme-Linked Immunosorbent Assay Based on Soluble Promastigote Antigen Detects Immunoglobulin M (IgM) and IgG Antibodies in Sera from Cases of Visceral and Cutaneous Leishmaniasis

    Science.gov (United States)

    Ryan, Jeffrey R.; Smithyman, Anthony M.; Rajasekariah, G-Halli; Hochberg, Lisa; Stiteler, John M.; Martin, Samuel K.

    2002-01-01

    Leishmaniasis causes significant morbidity and mortality in areas where it is endemic. In areas where it is nonendemic, global travel and increased incidence of the disease in human immunodeficiency virus and intravenous-drug user populations are also causes for concern. The unavailability of rapid and reliable tests for diagnosis of the various leishmaniases makes patient management difficult. We have developed an enzyme-linked immunosorbent assay (ELISA) that can detect immunoglobulin M (IgM) and IgG antibodies in patients with visceral and cutaneous leishmaniasis. These practical assays are based on soluble antigens from promastigotes cultivated in a protein-free medium. In preliminary studies, 129 visceral (Brazil, Italy, North Africa, and Nepal) and 143 cutaneous (Brazil) leishmaniasis patients with controls were tested. Overall, the tests showed a sensitivity of 95.1%. In addition, the ELISA correctly identified 42 sera from Brazilian dogs with canine leishmaniasis and 10 healthy controls. Serological tests for the various clinical manifestations of leishmaniasis could be useful epidemiological and patient management tools in populations of areas of endemicity and nonendemicity. PMID:11880434

  2. Immunoglobulin M and G antibody responses to Plasmodium falciparum glutamate-rich protein

    DEFF Research Database (Denmark)

    Dziegiel, Morten Hanefeld; Rowe, P; Bennett, S

    1993-01-01

    The aims of the present study were to describe the age-related immunoglobulin M (IgM) and IgG response to part of a 220-kDa glutamate-rich protein (GLURP) from Plasmodium falciparum and to determine possible correlations of possession of these antibodies with malaria morbidity. IgM and IgG levels...

  3. Correlation between specific IgM levels and percentage IgG-class antibody avidity to Toxoplasma gondii Correlação entre n��veis de IgM específica e percentual de avidez de anticorpos da classe IgG ao Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Marcel Leite

    2008-08-01

    Full Text Available Toxoplasmosis is an usually asymptomatic worldwide disseminated infection. In its congenital presentation it may lead to abortion or fetal malformations. Antenatal evaluation is considered of paramount importance to identify seronegative women and allow for prophylaxis. Recent improvements in sensitivity of IgM tests has made IgM detection an extremely protracted acute phase marker, and IgG avidity evaluation test became necessary. Observation has shown that a correlation can be established between IgM levels and avidity percentages, suggesting that frequently the avidity test may not be necessary. In this study we analyzed Toxoplasma gondii IgM levels of 202 samples and their IgG avidity percentages, in order to define specific levels whose IgM quantification could by itself define serodiagnosis and therefore make the avidity evaluation unnecessary. We showed that for IgM levels bellow 2.0 and above 6.0 serodiagnosis of toxoplasmosis could be established without need of IgG avidity test. IgM levels between these two parameters are associated with varying avidity indexes highlighting the importance of its evaluation as a means to confirm toxoplasmosis. Following this demonstration it was possible to avoid the avidity test for 75% of the cases, to reduce the turnaround time and to reduce costs.A Toxoplasmose é uma infecção universal e usualmente assintomática. A forma congênita, entretanto, pode resultar em aborto ou mal formações. Testes sorológicos estão indicados em situações onde há suspeita clínica, e na triagem pré-natal, quando são extremamente importantes para rastrear a infecção e orientar a gestante. O aumento da sensibilidade das técnicas para detecção de IgM, tornou necessário o desenvolvimento de recursos, como a avidez de IgG, visando obter novo marcador de infecção aguda. Embora exista correlação entre níveis de IgM e grau de avidez de IgG, a maioria dos testes de avidez associa-se a níveis baixos de IgM

  4. Antiphospholipid Antibodies in Women Undergoing In Vitro Fertilization Treatment: Clinical Value of IgA Anti-β2glycoprotein I Antibodies Determination

    Directory of Open Access Journals (Sweden)

    Odile Paulmyer-Lacroix

    2014-01-01

    Full Text Available Implantation failure could be related to antiphospholipid antibodies (aPL. We retrospectively analyzed the usefulness of aPL determination in women undergoing IVF. Conventional aPL of the antiphospholipid syndrome, lupus anticoagulant (LA, anticardiolipin antibodies (aCL, anti-β2glycoprotein I (aβ2GPI antibodies, and IgG and IgM isotypes as well as IgA isotype were analyzed in women presenting with at least two implantation failures after in vitro fertilization (IVF. In a population of 40 IVF patients, a total prevalence of 20% (8/40 of aPL was found, significantly different from that of the control population (100 healthy blood donors, P<0.0005. Among the panels of aPL tested, aβ2GPI IgA antibodies were the most prevalent (62.5% 5/8, significantly higher in IVF patients (12.5%, 5/40 than in controls (1%, 1/100 (P=0.01. No difference according to the numbers of IVF attempts and success of embryo implantation was found between aPL positive and negative IVF patients. In contrast, no accomplished pregnancy with full-term live birth was observed in aPL positive IVF patients. Altogether our data led us to propose aPL assessment, in particular aβ2GPI IgA antibodies, in support of IVF treated women. In a perspective way, an early aPL detection could be the basis for defining novel therapeutic strategy.

  5. IgM Augments Complement Bactericidal Activity with Serum from a Patient with a Novel CD79a Mutation

    NARCIS (Netherlands)

    J.D. Langereis (Jeroen); S.S.V. Henriet (Stefanie S.V.); Kuipers, S. (Saskia); C.M.R. Weemaes (Corry); M. van der Burg (Mirjam); M.I. de Jonge (Marien); M. van der Flier (Michiel)

    2018-01-01

    textabstractAntibody replacement therapy for patients with antibody deficiencies contains only IgG. As a result, concurrent IgM and IgA deficiency present in a large proportion of antibody deficient patients persists. Especially patients with IgM deficiency remain at risk for recurrent infections of

  6. Determining the Phagocytic Activity of Clinical Antibody Samples

    Science.gov (United States)

    McAndrew, Elizabeth G.; Dugast, Anne-Sophie; Licht, Anna F.; Eusebio, Justin R.; Alter, Galit; Ackerman, Margaret E.

    2011-01-01

    include phagocytic activity, cytokine secretion, and patterns of FcγRs usage, and are determined in a standardized manner, making this a highly useful system for parsing differences in this antibody-dependent effector function in both infection and vaccine-mediated protection9. PMID:22143444

  7. Monoclonal antibodies to equine CD23 identify the low-affinity receptor for IgE on subpopulations of IgM+ and IgG1+ B-cells in horses.

    Science.gov (United States)

    Wagner, Bettina; Hillegas, Julia M; Babasyan, Susanna

    2012-04-15

    CD23, also called FcεRII, is the low-affinity receptor for IgE and has first been described as a major receptor regulating IgE responses. In addition, CD23 also binds to CD21, integrins and MHC class II molecules and thus has a much wider functional role in immune regulation ranging from involvement in antigen-presentation to multiple cytokine-like functions of soluble CD23. The role of CD23 during immune responses of the horse is less well understood. Here, we expressed equine CD23 in mammalian cells using a novel IL-4 expression system. Expression resulted in high yield of recombinant IL-4/CD23 fusion protein which was purified and used for the generation of monoclonal antibodies (mAbs) to equine CD23. Seven anti-CD23 mAbs were further characterized. The expression of the low-affinity IgE receptor on equine peripheral blood mononuclear cells was analyzed by flow cytometric analysis. Cell surface staining showed that CD23 is mainly expressed by a subpopulation of equine B-cells. Only a very few equine T-cells or monocytes expressed CD23. CD23(+) B-cells were either IgM(+) or IgG1(+) cells. All CD23(+) cells were also positive for cell surface IgE staining suggesting in vivo IgE binding by the receptor. Two of the CD23 mAbs detected either the complete extracellular region of CD23 or a 22kDa cleavage product of CD23 by Western blotting. The new anti-CD23 mAbs provide valuable reagents to further analyze the roles of CD23 during immune responses of the horse in health and disease. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Heterotypic reactions in a radioimmunoassay for coxsackie B virus specific IgM.

    Science.gov (United States)

    Pugh, S F

    1984-01-01

    IgM antibody capture radioimmunoassays were developed to detect coxsackie virus B1-B5 specific IgM. Specific IgM was detected in sera from all patients with coxsackie B virus infections proved by isolation; however, sera from 13/32 patients with rising neutralising antibody titres were negative in the assay. Frequent heterotypic responses were seen among the positive sera. Thirty seven patients with other enterovirus infections were also studied, and sera from 15 of these patients reacted in the assay, showing that heterotypic coxsackie B IgM responses occur not only in coxsackie B virus infections but also in other enterovirus infections. PMID:6323548

  9. Determining the limit of detection of surface bound antibody.

    Science.gov (United States)

    Madiona, Robert M T; Welch, Nicholas G; Scoble, Judith A; Muir, Benjamin W; Pigram, Paul J

    2017-08-11

    Determination of a limit of detection (LoD) for surface bound antibodies is crucial for the development and deployment of sensitive bioassays. The measurement of very low concentrations of surface bound antibodies is also important in the manufacturing of pharmaceutical products such as antibody-conjugated pharmaceuticals. Low concentrations are required to avoid an immune response from the target host. Enzyme-linked immunosorbent assay (ELISA), x-ray photoelectron spectroscopy (XPS), and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were used to determine the LoD for the surface bound antibody (antiepidermal growth factor receptor antibody) on silicon substrates. Antibody solution concentrations between 10 μg/ml and 1 ng/ml and a control (antibody-free buffer solution) were employed, and the detection performance of each technique was compared. For this system, the ELISA LoD was 100 ng/ml and the XPS LoD was 1 μg/ml, corresponding to an estimated surface concentration of 49  ± 7 ng/cm 2 using a 1 μg/ml solution. Due to the multivariate complexity of ToF-SIMS data, analysis was carried out using three different methods, peak ratio calculations, principal component analysis, and artificial neural network analysis. The use of multivariate analysis with this dataset offers an unbiased analytical approach based on the peaks selected from ToF-SIMS data. The results estimate a ToF-SIMS LoD between applied antibody concentrations of 10 and 100 ng/mL. For surface bound antibodies on a silicon substrate, the LoD is below an estimated surface concentration of 49 ng/cm 2 . The authors have determined the LoD for this system using ELISA, XPS, and ToF-SIMS with multivariate analyses, with ToF-SIMS offering an order of magnitude better detection over ELISA and 2 orders of magnitude better detection over XPS.

  10. Evaluation of IgG and IgM anti-cytomegalovirus antibodies in parturient women and their newborns in the municipality of Presidente Prudente and region, State of São Paulo Pesquisa de anticorpos IgG e IgM para citomegalovírus em parturientes e recém-natos do município de Presidente Prudente e região, Estado de São Paulo

    Directory of Open Access Journals (Sweden)

    Ageane Monteiro Oliveira

    2002-04-01

    Full Text Available Primary infection by Cytomegalovirus (CMV is usually followed by a persistent and/or recurrent infection. In most cases the infection is sub-clinical, but it can eventually be acute. This paper aims to evaluate by ELISA the frequency of IgG and IgM anti-CMV antibodies in 86 samples of parturient women and their newborns living in Presidente Prudente, SP, and vicinities. The samples were collected at Dr.Domingos Cerávolo University Hospital. Three parturient women (3.5% were negative for anti-CMV antibodies, and 83(96.5% were positive for IgG only. About the newborns, 83(96.5% were positive for IgG only, 2(2.3% where positive for both antibodies, and 1(1,2% was negative for both antibodies. Results showed a high frequency of IgG anti-CMV antibodies in parturient women and their newborns. Moreover, we detected a relative high frequency of CMV congenitally infected newborns in this population, which might have an important impact on the amount of children with neurological and other diseases due to the virus infectionA infecção primária pelo citomegalovírus (CMV é freqüentemente seguida por infecção persistente e/ou recorrente. Na grande maioria dos casos a infecção é subclínica, podendo ser aguda em certas condições. O objetivo deste trabalho foi determinar por ELISA a freqüência de anticorpos IgG e IgM para CMV, em 86 parturientes e seus recém-natos (RNs provenientes do Hospital Universitário Dr. Domingos Cerávolo, Presidente Prudente, Estado de São Paulo. Os dados demonstraram que 83 parturientes (96,5% apresentaram anticorpos IgG anti-CMV e 3(3,5% não apresentaram anticorpos anti-CMV. Quanto aos RNs, 1(1,2% não apresentou anticorpos anti-CMV, 83(96,5% apresentaram anticorpos IgG e 2(2,3% apresentaram anticorpos IgM, caracterizando infecção aguda. Verificamos, assim, uma alta freqüência de anticorpos IgG para CMV em gestantes e RNs, bem como uma relativa alta freqüência de RNs infectados congenitamente pelo CMV nessa

  11. Early detection of leprosy by examination of household contacts, determination of serum anti-PGL-1 antibodies and consanguinity

    Directory of Open Access Journals (Sweden)

    Renata Bazan-Furini

    2011-08-01

    Full Text Available A cross-sectional clinical trial in which the serum anti-phenolic glycolipid (anti-PGL-1 antibodies were analysed in household contacts (HHC of patients with leprosy as an adjunct early leprosy diagnostic marker was conducted. The families of 83 patients underwent clinical examination and serum anti-PGL1 measurement using enzyme-linked immunosorbent assay. Of 320 HHC, 98 were contacts of lepromatous leprosy (LL, 80 were contacts of borderline lepromatous (BL, 28 were contacts of borderline (BB leprosy, 54 were contacts of borderline tuberculoid (BT, 40 were contacts of tuberculoid (TT and 20 were contacts of indeterminate (I leprosy. Consanguinity with the patients was determined for 232 (72.5% HHC. Of those 232 contacts, 183 had linear consanguinity. Forty-nine HHC had collateral consanguinity. Fifty-eight contacts (18.1% tested positive for anti-PGL1 antibodies. The number of seropositive contacts based on the clinical forms of the index case was 17 (29.3% for LL, 15 (25.9% for BL, one (1.7% for BB, 14 (24.1% for BT, three (5.2% for TT and eight (13.7% for I. At the one year follow-up, two (3.4% of these seropositive contacts had developed BT leprosy. The results of the present study indicate that the serum anti-PGL-1 IgM antibody may be useful for evaluating antigen exposure and as a tool for an early leprosy diagnosis in HHC.

  12. Raman spectroscopy-based screening of IgM positive and negative sera for dengue virus infection

    Science.gov (United States)

    Bilal, M.; Saleem, M.; Bilal, Maria; Ijaz, T.; Khan, Saranjam; Ullah, Rahat; Raza, A.; Khurram, M.; Akram, W.; Ahmed, M.

    2016-11-01

    A statistical method based on Raman spectroscopy for the screening of immunoglobulin M (IgM) in dengue virus (DENV) infected human sera is presented. In total, 108 sera samples were collected and their antibody indexes (AI) for IgM were determined through enzyme-linked immunosorbent assay (ELISA). Raman spectra of these samples were acquired using a 785 nm wavelength excitation laser. Seventy-eight Raman spectra were selected randomly and unbiasedly for the development of a statistical model using partial least square (PLS) regression, while the remaining 30 were used for testing the developed model. An R-square (r 2) value of 0.929 was determined using the leave-one-sample-out (LOO) cross validation method, showing the validity of this model. It considers all molecular changes related to IgM concentration, and describes their role in infection. A graphical user interface (GUI) platform has been developed to run a developed multivariate model for the prediction of AI of IgM for blindly tested samples, and an excellent agreement has been found between model predicted and clinically determined values. Parameters like sensitivity, specificity, accuracy, and area under receiver operator characteristic (ROC) curve for these tested samples are also reported to visualize model performance.

  13. Polyclonal antibody to ovomucoid determination in gamma irradiated laying eggs

    International Nuclear Information System (INIS)

    Harder, Marcia N.C.; Arthur, Valter; Silva, Lucia C.A.S.; Lopes, Tatiana G.G.; Duarte, Keila M.R.; Canniatti-Brazaca, Solange G.; Savino, Vicente J.M.; Coelho, Antonio A.D.

    2009-01-01

    To determine allergenic food proteins, one of the most used tests is the immunoassays such as ELISA (enzyme linked immunosorbent assay), where the antibody recognizes the antigen and this connection is showed by an enzymatic system, in other words, optical density. The aim of this study was to determine the polyclonal antibody efficiency, produced in laboratory, to identify the presence the ovomucoid antigen in treated eggs by gamma irradiation for its inactivation. To evaluate the treatments, polyclonal antibody was produced in female rabbits immunized with bioconjugated ovomucoid. Was used Freund Complete Adjuvant at first immunization and PBS Buffer at four subsequently immunizations every fifteen days, plus a booster 48 hours before the blood retreated. The blood serum was tittered by PTA-ELISA (Plate trapped antigen). All procedures were according to European Norms for ethical and animal welfare. It was used, in nature, commercial laying eggs. So the samples were submitted to the gamma radiation coming from a source of Co 60 , type Multipurpose, under a dose rate of 19.4 and 31.8 Gy/hour, in the doses: 0 (control); 10 KGy; 20 KGy and 30 KGy, in all rates. By the ELISA.s test we can find the egg allergen ovomucoid and the radiation treatment do not showed considerable changes. So we can concluded that the antibody produced is capable of identify the ovomucoid allergenic protein and the gamma irradiation in such rates does not shows changes in that protein, therefore showed some changes in the color and visual viscosity of the egg samples. (author)

  14. Selective IgM Deficiency—An Underestimated Primary Immunodeficiency

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    Sudhir Gupta

    2017-09-01

    Full Text Available Although selective IgM deficiency (SIGMD was described almost five decades ago, it was largely ignored as a primary immunodeficiency. SIGMD is defined as serum IgM levels below two SD of mean with normal serum IgG and IgA. It appears to be more common than originally realized. SIGMD is observed in both children and adults. Patients with SIGMD may be asymptomatic; however, approximately 80% of patients with SIGMD present with infections with bacteria, viruses, fungi, and protozoa. There is an increased frequency of allergic and autoimmune diseases in SIGMD. A number of B cell subset abnormalities have been reported and impaired specific antibodies to Streptococcus pneumoniae responses are observed in more than 45% of cases. Innate immunity, T cells, T cell subsets, and T cell functions are essentially normal. The pathogenesis of SIGMD remains unclear. Mice selectively deficient in secreted IgM are also unable to control infections from bacterial, viral, and fungal pathogens, and develop autoimmunity. Immunological and clinical similarities and differences between mouse models of deficiency of secreted IgM and humans with SIGMD have been discussed. Patients with SIGMD presenting with recurrent infections and specific antibody deficiency responses appear to improve clinically on immunoglobulin therapy.

  15. The neutralizing role of IgM during early Chikungunya virus infection.

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    Chong-Long Chua

    Full Text Available The antibody isotype IgM appears earlier than IgG, within days of onset of symptoms, and is important during the early stages of the adaptive immune response. Little is known about the functional role of IgM during infection with chikungunya virus (CHIKV, a recently reemerging arbovirus that has caused large global outbreaks. In this study, we studied antibody responses in 102 serum samples collected during CHIKV outbreaks in Malaysia. We described the neutralizing role of IgM at different times post-infection and examined the independent contributions of IgM and IgG towards the neutralizing capacity of human immune sera during the early phase of infection, including the differences in targets of neutralizing epitopes. Neutralizing IgM starts to appear as early as day 4 of symptoms, and their appearance from day 6 is associated with a reduction in viremia. IgM acts in a complementary manner with the early IgG, but plays the main neutralizing role up to a point between days 4 and 10 which varies between individuals. After this point, total neutralizing capacity is attributable almost entirely to the robust neutralizing IgG response. IgM preferentially binds and targets epitopes on the CHIKV surface E1-E2 glycoproteins, rather than individual E1 or E2. These findings provide insight into the early antibody responses to CHIKV, and have implications for design of diagnostic serological assays.

  16. The prevalence of toxoplasma igG and IgM in pregnant women residing in Rawalpindi

    International Nuclear Information System (INIS)

    Kausar, N.; Akhtar, S.; Ikram, A.

    2010-01-01

    Objective: To determine the prevalence of Toxoplasma gondii antibodies (IgG/IgM) among pregnant women visiting Military Hospital Rawalpindi and to develop a relationship between various risk factors and disease prevalence. Methods: One thousand pregnant women reporting in out patient Gynaecology department of Military Hospital (MH) Rawalpindi from October 2008 through January 2009 for antenatal check up were included in the study. Their serum samples were tested for the presence of Toxoplasma IgM and IgG immunoglobulins. Enzyme Linked immunosorbent assay test kits for both IgG and IgM were used to detect 1: gondii immunoglobulins in serum samples. Rest of the serum was stored at -20 degree C. Results: Of the 1000 women sampled at hospital, 46 (4.6%) had evidence of past infection and were seropositive for immunoglobulins of T. gondii IgG, while none of them were seropositive for IgM immunoglobulin, suggesting absence of recent infections during pregnancy. Conclusion: In twin cities of Islamabad and Rawalpindi, the sero prevalence of T. gondii IgG in pregnant women is relatively high (4.6%) as compared to other areas nearby. Consequently, the risk of, primary infection during pregnancy and the potential for congenital infection of foetus remains there as a large number of pregnant women were sero-negative for both the antibodies.

  17. Structural definition by antibody engineering of an idiotypic determinant.

    Science.gov (United States)

    Sollazzo, M; Castiglia, D; Billetta, R; Tramontano, A; Zanetti, M

    1990-05-01

    Using computer-aided techniques for predicting molecular structure, we constructed an atomic model of the variable domain of a murine anti-thyroglobulin antibody whose immunodominant idiotypic determinant (Id62) was mapped by site-directed mutagenesis and immunochemical analysis. We previously showed that under experimental conditions this idiotype activates anti-idiotypic B cells and T cells, and modulates the response to thyroglobulin in mice. Because idiotype interactions are considered of physiological importance for immune regulation, we studied this idiotype as a model to understand the relationship between function and structure. To determine the contribution of heavy- and light-chain variable domains to the idiotype structure, we constructed chimeric expression vectors and introduced them into the (non-secreting) P3X63Ag8.653 myeloma cell line. Mutants of the heavy-chain variable domain were obtained by site-directed mutagenesis and transfected into the murine (lambda 1) light-chain producer J558L cell line. The expressed proteins were purified from culture supernatants of transfected cells and characterized. We provide evidence that the third hypervariable loop (D region) of the heavy-chain variable domain is the structural correlate of the idiotypic determinant of this autoantibody and is independent from the nature of the associated light chain. Substitution of residues of the first and second complementarity-determining regions do not affect idiotype expression. The results described here are discussed in relation to our understanding, at a molecular level, of the interaction of idiotopes with B- and T-cell compartments.

  18. Impact of the Yosemite hantavirus outbreak on hantavirus antibody testing at a national reference laboratory.

    Science.gov (United States)

    Prince, Harry E; Lieberman, Jay M

    2013-08-01

    In conjunction with the 2012 Yosemite hantavirus outbreak, the number of sera our facility tested for hantavirus antibodies increased. We tracked test results and used the data set to determine if a more efficient testing algorithm was possible. Sera were screened using laboratory-developed pan-hantavirus IgG and IgM enzyme immunoassays (EIAs), with an index of >1.10 defined as positive. Sera that were IgM positive by screening (screen IgM(+)) were tested for Sin Nombre virus (SNV)-specific IgM using a laboratory-developed EIA; screen IgM(+) IgG(+) sera were also tested for SNV IgG using a laboratory-developed immunoblot assay. SNV antibody-positive samples were sent to state public health laboratories (PHL) or the CDC for confirmation. Of 3,946 sera tested from July through December 2012, 205 were screen IgM(+) IgG negative (IgG(-)); 7/205 were SNV IgM(+), but only 1/5 sent to PHL/CDC was confirmed as SNV IgM(+). Of 61 screen IgM(+) IgG(+) sera, 16 were SNV antibody positive; 13/16 sera (from 11 patients) went to PHL/CDC, where SNV infection was confirmed for all patients. Of 12 confirmed patients, 7 had been exposed at Yosemite. A modified algorithm defining screen indices of ≥2.00 as positive identified 11/12 confirmed cases while reducing the number of sera requiring SNV-specific antibody testing by 65%; the patient missed was not tested until 3 months after the onset of symptoms. Hantavirus antibody testing at our facility identified 12 SNV-infected patients, including 7 exposed at Yosemite. Some screen IgM(+) IgG(-) SNV IgM(+) results were false positives, emphasizing the value of PHL/CDC confirmatory testing. We identified a modified algorithm requiring analysis of fewer specimens for SNV-specific antibodies without loss of sensitivity.

  19. Emerging functions of natural IgM and its Fc receptor FCMR in immune homeostasis

    Directory of Open Access Journals (Sweden)

    Hongsheng eWang

    2016-03-01

    Full Text Available Most natural IgM antibodies are encoded by germline Ig sequences and are produced in large quantities by both mice and humans in the absence of intentional immunization. Natural IgM are reactive with many conserved epitopes, including those shared by microorganisms and autoantigens. As a result, these antibodies play important roles in clearing intruding pathogens, as well as apoptotic/necrotic cells and otherwise damaged tissues. While natural IgM binds to target structures with low affinity due to a lack of significant selection by somatic hypermutation, its pentameric structure with 10 antigen binding sites enables these antibodies to bind multivalent target antigens with high avidity. Opsonization of antigen complexed with IgM is mediated by cell surface Fc receptors. While the existence of Fc alpha/mu receptor has been known for some time, only recently has the Fc receptor specific for IgM (FCMR been identified. In this review, we focus on our current understandings of how natural IgM and FCMR regulate the immune system and maintain homeostasis under physiological and pathological conditions.

  20. Characterization of the antibody response to a Haemophilus influenzae type b conjugate vaccine in children with recurrent lower respiratory tract infection

    DEFF Research Database (Denmark)

    Kristensen, K; Barington, T; Pressler, T

    1995-01-01

    once with a Haemophilus influenzae type b (Hib) conjugate vaccine. Total IgG subclasses, total antipolysaccharide Hib antibodies, and antipolysaccharide Hib antibodies of IgM, IgG, IgA, and IgG1-4 specificity were determined by ELISA. There were no significant differences between the two groups in any...

  1. Serological profile of HSV-2 in patients attending STI clinic: Evaluation of diagnostic utility of HSV-2 IgM detection

    Directory of Open Access Journals (Sweden)

    Choudhry Shilpee

    2009-07-01

    Full Text Available Objective: The present study was done to evaluate the serological profile of herpes simplex virus-2 (HSV-2 among patients attending sexually transmitted infections (STI clinic and to determine the utility of detecting HSV-2 IgM antibodies in such patients. A correlation of HSV-2 infection with other STI including HIV has also been attempted. Materials and Methods: Hundred consecutive patients who attended STI clinic, with one or more of the complaints as enunciated by WHO in syndromic approach for the diagnosis of STI, were included as subjects. All subjects were screened for common STI by standard laboratory procedures/ commercially available kits. HSV-1 and HSV-2 IgM antibody was detected by commercially available enzyme immuno assay kit in all patient′s sera. Sera were also tested for other STI, namely HIV, Hepatitis B virus, Hepatitis C virus and Treponema pallidum. Antigen detection for Chlamydia trachomatis was done in genital swabs of all patients by Bio-Rad Chlamydia Microplate EIA 31189 (United States kit. Results: Thirty patients were found to have genital herpes. In 17/30 (56.6% patients, HSV-2 serology was found to correlate with the clinical diagnosis. The coexistence of other infection in HSV-2 seropositive patients was detected in 8/30 patients. None of the patients having concomitant infections were clinically diagnosed accurately. Sensitivity, specificity, positive predictive value and negative predictive value of IgM antibodies for the diagnosis of genital herpes was 73.91%, 90.91%, 70.83% and 92.91% respectively. Conclusion: HSV-2 IgM detection could only be used as a supportive test for the diagnosis of genital herpes . It needs to be emphasized that the sensitivity and positive predictive value scores are pointers for further improvement in the commercial assay systems and a large sample size may determine the broader utility of such systems.

  2. Determination of Antibody Titres and Isolation of Salmonella species ...

    African Journals Online (AJOL)

    A total of 1,016 patients attending the University of Calabar Medical Centre with symptoms of fever resembling typhoid and enteric fever were used for this study. Fifty-five healthy controls from the University Community were also studied. Serum samples from patients and controls were screened for Salmonella antibodies ...

  3. Recognition of Porphyromonas gingivalis gingipain epitopes by natural IgM binding to malondialdehyde modified low-density lipoprotein.

    Directory of Open Access Journals (Sweden)

    S Pauliina Turunen

    Full Text Available OBJECTIVE: Increased risk for atherosclerosis is associated with infectious diseases including periodontitis. Natural IgM antibodies recognize pathogen-associated molecular patterns on bacteria, and oxidized lipid and protein epitopes on low-density lipoprotein (LDL and apoptotic cells. We aimed to identify epitopes on periodontal pathogen Porphyromonas gingivalis recognized by natural IgM binding to malondialdehyde (MDA modified LDL. METHODS AND RESULTS: Mouse monoclonal IgM (MDmAb specific for MDA-LDL recognized epitopes on P. gingivalis on flow cytometry and chemiluminescence immunoassays. Immunization of C57BL/6 mice with P. gingivalis induced IgM, but not IgG, immune response to MDA-LDL and apoptotic cells. Immunization of LDLR(-/- mice with P. gingivalis induced IgM, but not IgG, immune response to MDA-LDL and diminished aortic lipid deposition. On Western blot MDmAb bound to P. gingivalis fragments identified as arginine-specific gingipain (Rgp by mass spectrometry. Recombinant domains of Rgp produced in E. coli were devoid of phosphocholine epitopes but contained epitopes recognized by MDmAb and human serum IgM. Serum IgM levels to P. gingivalis were associated with anti-MDA-LDL levels in humans. CONCLUSION: Gingipain of P. gingivalis is recognized by natural IgM and shares molecular identity with epitopes on MDA-LDL. These findings suggest a role for natural antibodies in the pathogenesis of two related inflammatory diseases, atherosclerosis and periodontitis.

  4. Modulation of immune response by bacterial lipopolysaccharide (LPS): cellular basis of stimulatory and inhibitory effects of LPS on the in vitro IGM antibody response to a T-dependent antigen

    International Nuclear Information System (INIS)

    Uchiyama, T.; Jacobs, D.M.

    1978-01-01

    The role of thymus-derived lymphocytes (T cells) in LPS modulation of T cell-development antibody responses has been investigated. We have assessed the effect of LPS on the primary anti-TNP response to TNP-SRBC of cultures of whole spleen cells or T cell-depleted spleen cells that were supplemented with various subpopulations of carrier-primed (SRBC) spleen cells. The TNP-PFC response was enhanced in the presence of irradiated SRBC-primed spleen cells by addition of 0.16 to 20 μg/ml LPS, but inhibition was observed when irradiation of primed cells was omitted. Enhancement but no inhibition occurred when added primed cells were first passed through a nylon wool column. LPS-mediated enhancement was dependent on a T cell in the primed population. These results suggest that LPS modulation of antibody synthesis is dependent on two populations of antigen-specific cells that have opposing effects on B cell responses to a T-dependent antigen: a helper cell that is irradiation resistant, nonadherent to nylon wool, and sensitive to anti-T cell serum, and a suppressor cell that is irradiation sensitive and adherent to nylon wool

  5. Determination of specific IgG antibody by crossed radioimmunoelectrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Nordvall, S.L. (Dept. of Paediatrics, University Hospital, Uppsala, Sweden); Uhlin, T.; Einarsson, R. (Allergy Research, Pharmacia Diagnostics AB, Uppsala, Sweden)

    1983-01-01

    A crossed radioimmunoelectrophoretic method was developed for detection of honey bee venom specific IgG antibodies in patient sera. At the serum concentration 1/200 the contrast between specific binding and backgroud was the most favourable. The detection limit was fairly low, approximately 30 kU/l(IgG RAST units). A reference system based on the reference kits in Phadebas IgG-RAST was elaborated.

  6. Determination of specific IgG antibody by crossed radioimmunoelectrophoresis

    International Nuclear Information System (INIS)

    Nordvall, S.L.; Uhlin, T.; Einarsson, R.

    1983-01-01

    A crossed radioimmunoelectrophoretic method was developed for detection of honey bee venom specific IgG antibodies in patient sera. At the serum concentration 1/200 the contrast between specific binding and backgroud was the most favourable. The detection limit was fairly low, approximately 30 kU/l(IgG RAST units). A reference system based on the reference kits in Phadebas IgG-RAST was elaborated. (author)

  7. The new ParaDIgm: IgM from bench to clinic: November 15-16, 2011, Frankfurt, Germany.

    Science.gov (United States)

    Hanala, Sherif

    2012-01-01

    The inaugural IgM event entitled "The new ParaDIgm: IgM from bench to clinic" brought together the increasingly active and growing IgM antibody community to discuss recent advances and challenges facing the discovery and development of IgM antibody therapies and technologies. Researchers, clinicians and biomanufacturing experts delivered 21 talks on the basic science and isolation of IgM, upstream and downstream development, and formulation and clinical development of the molecules. Participants networked around topics aimed at exploring the full potential of IgM antibodies. The meeting was held at DECHEMA Gesellschaft für Chemische Technik und Biotechnologie e. V. (Society for Chemical Engineering and Biotechnology), a non-profit scientific and technical society based in Frankfurt am Main, Germany. The meeting was sponsored by Patrys, Laureate Biopharma, Bio-Rad Laboratories, BIA Separations, Percivia and the Bio Affinity Company (BAC). The second New ParaDIgm: IgM from bench to clinic meeting, will be held on April 23-24, 2013 in Frankfurt, Germany.

  8. Research advances in determination of hepatitis B core antibody level

    OpenAIRE

    WANG Wei; ZHAO Zhaoxia; LI Yuxiao

    2016-01-01

    With the development and application of the double antigen sandwich method for quantification of hepatitis B core antibody (HBcAb) in recent years, there is increasing knowledge of the ability of HBcAb to reflect the body′s anti-viral capability. This article introduces the commonly used measurement methods for HBcAb and the new trends in HBcAb measurement and summarizes the association of serum HBcAb level with viral antigen and the body′s immune response, as well as research advances in eff...

  9. Shift in epitope dominance of IgM and IgG responses to Plasmodium falciparum MSP1 block 4

    Directory of Open Access Journals (Sweden)

    Leke Rose GF

    2010-01-01

    Full Text Available Abstract Background Plasmodium falciparum merozoite surface protein-1 (MSP1 has been extensively studied as a blood-stage malaria vaccine candidate, with most work focused on the conserved 19 kDa and semi-conserved 42 kDa C-terminal regions (blocks 16-17 and the hypervariable N-terminal repeat region (block 2. However, recent genotyping studies suggest that additional regions of MSP1 may be under selective pressure, including a locus of intragenic recombination designated as block 4 within the 3' region of the gene. Methods The current study examined the antibody response to the two parental and two recombinant forms of block 4 and to blocks 16-17 (3D7 in study populations from Colombia, Papua New Guinea and Cameroon that differ in malaria transmission intensity and ethnic composition. Results IgM and IgG antibodies were detected against parental and recombinant MSP1 block 4 peptides in all three populations. Overall, 32-44% of the individuals produced IgM to one or more of the peptides, with most individuals having IgM antibodies reactive with both parental and recombinant forms. In contrast, IgG seropositivity to block 4 varied among populations (range 15-65%, with the majority of antibodies showing specificity for one or a pair of block 4 peptides. The IgG response to block 4 was significantly lower than that to blocks 16-17, indicating block 4 is subdominant. Antibodies to block 4 and blocks 16-17 displayed distinct IgG subclass biases, with block 4 responses biased toward IgG3 and blocks 16-17 toward IgG1. These patterns of responsiveness were consistently observed in the three study populations. Conclusions Production of antibodies specific for each parental and recombinant MSP1 block 4 allele in different populations exposed to P. falciparum is consistent with balancing selection of the MSP1 block 4 region by the immune response of individuals in areas of both low and high malaria transmission. MSP1 block 4 determinants may be important

  10. Salmonella Challenge Affects the Antibody Isotype Profile of Bile in Hens Differing in Metabolic Efficiency

    NARCIS (Netherlands)

    Cotter, P.F.; Eerden, van E.

    2006-01-01

    Gel precipitation reactions determined antibody isotypes in bile from hens differing in dietary efficiency. Ouchterlony double diffusion employing ¿-chain specific goat-anti-chicken IgA, rabbit anti-chicken IgG, goat anti-chicken IgM, black turtle bean (BTB), and Jacalin lectins as precipitating

  11. Research advances in determination of hepatitis B core antibody level

    Directory of Open Access Journals (Sweden)

    WANG Wei

    2016-07-01

    Full Text Available With the development and application of the double antigen sandwich method for quantification of hepatitis B core antibody (HBcAb in recent years, there is increasing knowledge of the ability of HBcAb to reflect the body′s anti-viral capability. This article introduces the commonly used measurement methods for HBcAb and the new trends in HBcAb measurement and summarizes the association of serum HBcAb level with viral antigen and the body′s immune response, as well as research advances in effective prediction of antiviral effect with baseline HBcAb measurement before antiviral therapy. It is also pointed out that the clinical application of HBcAb needs further investigation.

  12. Simultaneous use of serum IgG and IgM for risk scoring of suspected early Lyme borreliosis: graphical and bivariate analyses

    DEFF Research Database (Denmark)

    Dessau, Ram B; Ejlertsen, Tove; Hilden, Jørgen

    2010-01-01

    The laboratory diagnosis of early disseminated Lyme borreliosis (LB) rests on IgM and IgG antibodies in serum. The purpose of this study was to refine the statistical interpretation of IgM and IgG by combining the diagnostic evidence provided by the two immunoglobulins and exploiting the whole...

  13. Frequency of cytomegalo virus igm and igg in pre renal transplant donors and recipients

    International Nuclear Information System (INIS)

    Ashraf, S.

    2014-01-01

    One of the most opportunistic infection in kidney transplant recipients is Cytomegalovirus. This study was conducted to find the frequency of cytomegalo- virus IgM and IgG in kidney transplant donors and recipients. Methods: We carried out retrospective analysis of laboratory values of cytomegalovirus IgG and IgM antibodies of all donors and recipients presented from 2006 to 2013 to National Institute of Kidney diseases Sheikh Zayed Hospital Lahore Pakistan. Results: Total no. of patients were 410 (205 donors and 205 recipients). Among recipients 169 (82%) were male and 36 (18%) female while among donors 125 (60%) were male and 80(40%) female. Total 05 (2.4 %) recipients were CMV IgG negative and 03 (1.4%) CMV IgM positive. Out of two hundred five recipients 200 (98%) were IgG positive and 202 (99%) were IgM negative. All the donors were CMV IgM negative and CMV IgG positive. Conclusion: There is very high frequency of CMV IgM and IgG positivity in donors and recipient of renal transplant in our population so we should consider prophylactic therapy. (author)

  14. Toxoplasma-IgM and IgG-avidity in single samples from areas with a high infection rate can determine the risk of mother-to-child transmission Avidez de IgM e IgG de Toxoplasma em amostras de áreas com alta taxa de infecção pode determinar risco de transmissão materno-fetal

    Directory of Open Access Journals (Sweden)

    Myrian Morussi Reis

    2006-04-01

    Full Text Available Anti-Toxoplasma IgG-avidity was determined in 168 serum samples from IgG- and IgM-positive pregnant women at various times during pregnancy, in order to evaluate the predictive value for risk of mother-to-child transmission in a single sample, taking the limitations of conventional serology into account. The neonatal IgM was considered the serologic marker of transmission. Fluorometric tests for IgG, IgM (immunocapture and IgG-avidity were performed. Fifty-one of the 128 pregnant women tested gave birth in the hospital and neonatal IgM was obtained. The results showed 32 (62.75% pregnant women having high avidity, IgM indexes between 0.6 and 2.4, and no infected newborn. Nineteen (37.25% had low or inconclusive avidity, IgM indexes between 0.6 and 11.9, and five infected newborns and one stillbirth. In two infected newborns and the stillbirth maternal IgM indexes were low and in one infected newborn the only maternal parameter that suggested fetal risk was IgG-avidity. In the present study, IgG-avidity performed in single samples from positive IgM pregnant women helped to determine the risk of transmission at any time during pregnancy, especially when the indexes of the two tests were analysed with respect to gestational age. This model may be less expensive in developing countries where there is a high prevalence of infection than the follow-up of susceptible mothers until childbirth with monthly serology, and it creates a new perspective for the diagnosis of congenital toxoplasmosis.A avidez de IgG anti-Toxoplasma foi realizada em 168 amostras IgG e IgM positivas de gestantes, coletadas em qualquer período da gestação, para avaliar o valor preditivo do risco de transmissão materno-fetal em amostra única, considerando as limitações da sorologia convencional. A IgM neonatal foi considerada o marcador sorológico de transmissão. Testes fluorométricos foram realizados para IgG, IgM (imunocaptura e avidez de IgG. Cinqüenta e uma das 128

  15. Rationalization and Design of the Complementarity Determining Region Sequences in an Antibody-Antigen Recognition Interface

    Science.gov (United States)

    Chen, Ing-Chien; Lee, Yu-Ching; Chen, Jun-Bo; Tsai, Keng-Chang; Chen, Ching-Tai; Chang, Jeng-Yih; Yang, Ei-Wen; Hsu, Po-Chiang; Jian, Jhih-Wei; Hsu, Hung-Ju; Chang, Hung-Ju; Hsu, Wen-Lian; Huang, Kai-Fa; Ma, Alex Che; Yang, An-Suei

    2012-01-01

    Protein-protein interactions are critical determinants in biological systems. Engineered proteins binding to specific areas on protein surfaces could lead to therapeutics or diagnostics for treating diseases in humans. But designing epitope-specific protein-protein interactions with computational atomistic interaction free energy remains a difficult challenge. Here we show that, with the antibody-VEGF (vascular endothelial growth factor) interaction as a model system, the experimentally observed amino acid preferences in the antibody-antigen interface can be rationalized with 3-dimensional distributions of interacting atoms derived from the database of protein structures. Machine learning models established on the rationalization can be generalized to design amino acid preferences in antibody-antigen interfaces, for which the experimental validations are tractable with current high throughput synthetic antibody display technologies. Leave-one-out cross validation on the benchmark system yielded the accuracy, precision, recall (sensitivity) and specificity of the overall binary predictions to be 0.69, 0.45, 0.63, and 0.71 respectively, and the overall Matthews correlation coefficient of the 20 amino acid types in the 24 interface CDR positions was 0.312. The structure-based computational antibody design methodology was further tested with other antibodies binding to VEGF. The results indicate that the methodology could provide alternatives to the current antibody technologies based on animal immune systems in engineering therapeutic and diagnostic antibodies against predetermined antigen epitopes. PMID:22457753

  16. Padronização de ensaio imunoenzimático para pesquisa de anticorpos das classes IgM e IgG anti-Toxoplasma gondii e comparação com a técnica de imunofluorescência indireta Standardization of enzyme-linked immunosorbent assay ELISA to detect anti-Toxoplasma gondii IgM and IgG antibodies, and comparison with the indirect immunofluorescence technique

    Directory of Open Access Journals (Sweden)

    Cláudia Maria Antunes Uchôa

    1999-12-01

    Full Text Available A sorologia tem sido o método de escolha para o diagnóstico da toxoplasmose. Devido a isto, padronizamos um ensaio imunoenzimático (ELISA e comparamos seus resultados com a técnica de imunofluorescência indireta (IFI. A técnica padronizada apresentou na pesquisa de IgG sensibilidade (S de 96,7% e especificidade (E de 75%, com valor de predição de positividade (VPP de 83,3% e de negatividade (VPN de 94,7%, com uma concordância ajustada (K de 73,5%. A IFI apresentou S de 83,8%, E de 79,1% com VPP de 83,8 % e VPN de 79,1% com K de 63%. A concordância bruta entre os dois testes (ELISA/IFI foi de 88,3% para pesquisa de IgG e de 81,5% para pesquisa de IgM, sendo o K de 70,8% para IgG e de 1,3% para IgM, sendo o índice de correlação (r de 0,556 para IgG e de -0,023 para IgM. Podemos concluir que a ELISA-IgG padronizada é indicada nos processos de triagem sorológica, sendo a ELISA-IgM desaconselhada uma vez que apresentou baixos índices de concordância ajustada com a técnica de referência, sugerindo pouca confiabilidade dos resultados.Serology has been the most popular method to diagnose toxoplasmosis. Accordingly, this study standardizes an enzyme-linked immunosorbent assay (ELISA and compares its results with the IFI technique. In the IgG detection test, the standardized technique presented a sensibility (S of 96.77%, a specificity (SP of 75%, with a positive predictive value (PPV of 83.33%, a negative predictive value (NPV of 94.74%, and an adjusted concordance (K of 73.50%. The IFI exhibited 83.87% for S, 79.16% for SP, 83.81% for PPV, 79.16% for NPV, and 63% for K. The rough concordance between these two tests (ELISA/IFI was 88.35% for the IgG detection test and 81.55% for the IgM detection test. K was 70.82% and 1.31% for IgG and IgM, respectively, the correlation index (r being 0.556 for IgG and -0.023 for IgM. We can conclude that standardized ELISA-IgG is indicated in serologic selection processes, whereas the ELISA-IgM is

  17. Characterisation of monoclonal antibodies for human luteinising hormone, and mapping of antigenic determinants on the hormone

    International Nuclear Information System (INIS)

    Soos, M.; Siddle, K.

    1983-01-01

    Twelve mouse monoclonal antibodies for human luteinising hormone were produced. The affinities varied from 4 X 10 7 to 1 X 10 10 l/mol. The specificity of each antibody was assessed by determining the relative reactivities with luteinising hormone, thyroid stimulating hormone, follicle stimulating hormone and chorionic gonadotrophin. Six antibodies bound to the α-subunit as shown by similar reactivity with all hormones, and the remainder to the β-subunit as shown by specificity for luteinising hormone. This latter group of antibodies cross-reacted only weakly with thyroid stimulating hormone (approximately 10%) and follicle stimulating hormone (approximately 3%). Three of these antibodies also showed low reactivity towards chorionic gonadotrophin (<10%), though the others did not (80-300%). The ability of different antibodies to bind simultaneously to luteinising hormone was examined and it was shown that several distinct antigenic determinants existed on both subunits. The characterisation of monoclonal binding sites is discussed in relation to the use of antibodies in two-site immunoradiometric assays. (Auth.)

  18. Fatores associados com a soroprevalência de anticorpos IgG e IgM anti-toxoplasma em pessoas com HIV/AIDS atendidas em um serviço de assistência especializadaFactors associated with soroprevalence of antibodies IgG and IgM anti-toxoplasma in HIV/AIDS patients assisted in an expert assistance servicedoi:10.12662/2317-3076jhbs.v4i3.733.p145-151.2016

    Directory of Open Access Journals (Sweden)

    Joice Silva Alves

    2016-09-01

    Full Text Available Introdução: A AIDS é um dos principais problemas mundiais em nível de Saúde Pública. O indivíduo infectado pelo HIV torna-se mais suscetível a diversas infecções oportunistas, como a toxoplasmose e suas complicações potenciais, como a neurotoxoplasmose e a retinocoroidite. O entendimento do perfil sorológico Anti-toxoplasma e das características clínicas e laboratoriais de pessoas que vivem com HIV/AIDS é essencial para a elaboração de melhores alternativas de prevenção e de tratamento. Métodos: Foram analisados os dados de 547 pessoas com HIV/AIDS atendidas no Serviço de Assistência Especializada do município de Divinópolis–MG, de março de 2012 a janeiro de 2014. Por meio de modelos de regressão logística, foram pesquisados diferentes fatores associados com o perfil sorológico Anti-toxoplasma IgG e IgM dos participantes. Analisou-se, ainda, se as contagens de células T-CD4+ e se os valores de carga viral (CV diferiram de acordo com o resultado das sorologias. Resultados: As soroprevalências encontradas foram de 69% para IgG e de 6% para IgM. Observou-se que indivíduos solteiros e que não trabalhavam tiveram uma maior soropositividade para IgM. O não uso de TARV e uma menor escolaridade, por sua vez, foram associados com maiores prevalências de reatividade para IgG. Observou-se uma maior contagem de células T-CD4+ em indivíduos IgM positivos. Conclusão: O presente estudo identificou uma prevalência elevada de sorologias reagentes para a toxoplasmose em indivíduos com HIV/AIDS e apresentou variáveis a serem consideradas como prioritárias em ações de promoção e de prevenção.

  19. Identification of an immunodominant epitope in glycoproteins B and G of herpes simplex viruses (HSVs) using synthetic peptides as antigens in assay of antibodies to HSV in herpes simplex encephalitis patients.

    Science.gov (United States)

    Bhullar, S S; Chandak, N H; Baheti, N N; Purohit, H J; Taori, G M; Daginawala, H F; Kashyap, R S

    2014-01-01

    Herpes simplex encephalitis (HSE) is a severe viral infection of the central nervous system (CNS). Assay of antibody response is widely used in diagnostics of HSE. The aim of this study was to identify an immunodominant epitope determining the antibody response to herpes simplex viruses (HSVs) in cerebrospinal fluid (CSF) of HSE patients. The synthetic peptides that resembled type-common as well as type-specific domains of glycoproteins B (gB) and G (gG) of these viruses were evaluated for binding with IgM and IgG antibodies in CSF samples from HSE and non-HSE patients in ELISA. The QLHDLRF peptide, derived from gB of HSV was found to be an immunodominant epitope in the IgM and IgG antibody response. The patients with confirmed and suspected HSE showed in ELISA against this peptide 26% and 23% positivities for IgM, 43% and 37% positivities for IgG and 17% and 15% for both IgM and IgG antibodies, respectively. The total positivities of 86% and 75% for both IgM and IgG antibodies were obtained in the patients with confirmed and suspected HSE, respectively. These results demonstrate that a synthetic peptide-based diagnostics of HSE can be an efficient and easily accessible alternative. This is the first report describing the use of synthetic peptides derived from HSVs in diagnostics of HSE using patientsʹ CSF samples.

  20. Cross-Reactivity of Rapid Salmonella Typhi IgM Immunoassay in Dengue Fever Without Co-Existing Infection.

    Science.gov (United States)

    Bhatti, Adnan Bashir; Ali, Farhan; Satti, Siddique Akbar

    2015-12-04

    Dengue fever is endemic in developing nations worldwide with as many as 500,000 annual cases of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). A prompt and accurate diagnosis early in the disease course is essential for prompt identification and treatment of severe complications of the dengue virus infection (DVI). We identified cross-reactivity of a rapid IgM test for typhoid fever in patients with febrile illnesses that were determined to be due to dengue virus. All patients with documented DVI during a recent epidemic in Pakistan also underwent diagnostic testing for Salmonella enterica serovar Typhi. The diagnosis of DVI was made based on clinical findings and the positive results for dengue non-structural protein 1 antigen (NS1Ag) and/or dengue IgM antibody (anti-D IgM) during the acute phase of febrile illness. Patients with positive test results for Salmonella typhi (S. Typhi) IgM also had their blood cultures done. In the group of 322 patients with clinical and serological evidence of DVI, 107 also tested positive for S. Typhi IgM. Blood cultures were negative for S. Typhi bacteria in all patients. Principal disease features included fever, headache, myalgia, retro-orbital pain, and a rash accompanied by thrombocytopenia and leukopenia. Comparisons of clinical and routine laboratory findings between the S. Typhi-positive and negative groups showed no significant differences. Patients testing positive for both NS1Ag and anti-D IgM were significantly more likely to test positive for S. Typhi IgM, even in the absence of typhoid fever. No routine antibiotics were used and all patients survived. One-third of a large group of patients with primary DVI also demonstrated false positive results for typhoid fever. Cross-reactivity of a rapid immunoassay for typhoid fever has not been previously reported in DVI or any other flavivirus infections. Until these findings can be further evaluated, clinicians should be cautious in

  1. Selective IgM Deficiency: Clinical and Laboratory Features of 17 Patients and a Review of the Literature.

    Science.gov (United States)

    Chovancova, Zita; Kralickova, Pavlina; Pejchalova, Alena; Bloomfield, Marketa; Nechvatalova, Jana; Vlkova, Marcela; Litzman, Jiri

    2017-08-01

    Primary selective IgM deficiency (sIgMD) is a primary immunodeficiency with unclear pathogenesis and a low number of published cases. We reviewed clinical and laboratory manifestations of 17 sIgMD patients. Serum IgM, IgG, and its subclasses, IgA, IgE, antibodies against tetanus toxoid, pneumococcal polysaccharides and Haemophilus influenzae type b, isohemagglutinins, and T and B lymphocyte subsets, expressions of IgM on B cells and B lymphocyte production of IgM were compared with previously reported case reports and a small series of patients, which included 81 subjects in total. We found that some patients in our cohort (OC) and published cases (PC) had increased IgE levels (OC 7/15; PC 21/37), decreased IgG4 levels (OC 5/14), very low titers of isohemagglutinins (OC 8/8; PC 18/21), increased transitional B cell counts (OC 8/9), decreased marginal zone B cell counts (OC 8/9), and increased 21 low B cell counts (OC 7/9). Compared with the PC (20/20), only two of five OC patients showed very low or undetectable production of IgM after stimulation. A majority of the patients had normal antibody production to protein and polysaccharide antigens, basic lymphocyte subset counts, and expression of surface IgM molecules on B cells. Low IgM levels are associated with various immunopathological disorders; however, pathogenic mechanisms leading to decreased IgM serum level in selective IgM deficiency remain unclear. Moreover, it is difficult to elucidate how strong these associations are and if these immunopathological conditions are primary or secondary.

  2. Differential expression of IgE and IgG4 specific antibody responses in asymptomatic and chronic human filariasis

    NARCIS (Netherlands)

    Kurniawan, A.; Yazdanbakhsh, M.; van Ree, R.; Aalberse, R.; Selkirk, M. E.; Partono, F.; Maizels, R. M.

    1993-01-01

    A population of 164 adult individuals resident in an area endemic for Brugia malayi lymphatic filariasis has been studied for humoral immune responses to filarial parasites. Antibody levels to Ag extracted from adult worms were determined for each of the IgG subclasses, for IgM and for IgE. The

  3. High-throughput immunoturbidimetric assays for in-process determination of polyclonal antibody concentration and functionality in crude samples

    DEFF Research Database (Denmark)

    Bak, Hanne; Kyhse-Andersen, J.; Thomas, O.R.T.

    2007-01-01

    We present fast, simple immunoturbidimetric assays suitable for direct determination of antibody 'concentration' and 'functionality' in crude samples, such as in-process samples taken at various stages during antibody purification. Both assays display excellent linearity and analytical recovery. ...... antibodies, require only basic laboratory equipment, are robust, fast, cheap, easy to perform, and readily adapted to automation....

  4. Systemic lupus erythematosus patients contain significantly less igm against mono-methylated lysine than healthy subjects.

    Directory of Open Access Journals (Sweden)

    Sha Guo

    Full Text Available Post-translational modifications on proteins are important in biological processes but may create neo-epitopes that induce autoimmune responses. In this study, we measured the serum IgG and IgM response to a set of non-modified or acetyl- and methyl-modified peptides corresponding to residues 1-19 of the histone 3 N-terminal tail in systemic lupus erythematosus (SLE patients and healthy subjects. Our results indicated that the SLE patients and healthy subjects produced antibodies (Abs to the peptides, but the two groups had different Ab isotype and epitope preferences. Abs to the non-modified form, H31-19, were of the IgG isotype and produced by SLE patients. They could not recognize the scrambled H31-19, which contained the same amino acid composition but a different sequence as H31-19. In comparison, healthy subjects in general did not produce IgG against H31-19. However, about 70% of the healthy subjects produced IgM Abs against mono-methylated K9 of H31-19 (H31-19K9me. Our further studies revealed that ε-amine mono-methylated lysine could completely inhibit the IgM binding to H31-19K9me, but lysine had no inhibitory effect. In addition, the IgM Abs could bind peptides containing a mono-methylated lysine residue but with totally different sequences. Thus, mono-methylated lysine was the sole epitope for the IgM. Interestingly, SLE patients had much lower levels of this type of IgM. There was no obvious correlation between the IgM levels and disease activity and the decreased IgM was unlikely caused by medical treatments.We also found that the IgM Abs were not polyreactive to dsDNA, ssDNA, lipopolysaccharide (LPS or insulin and they did not exist in umbilical cord serum, implying that they were not natural Abs. The IgM Abs against mono-methylated lysine are present in healthy subjects but are significantly lower in SLE patients, suggesting a distinct origin of production and special physiological functions.

  5. A novel heavy domain antibody library with functionally optimized complementarity determining regions.

    Directory of Open Access Journals (Sweden)

    Ole Aalund Mandrup

    Full Text Available Today a number of synthetic antibody libraries of different formats have been created and used for the selection of a large number of recombinant antibodies. One of the determining factors for successful isolation of recombinant antibodies from libraries lies in the quality of the libraries i.e. the number of correctly folded, functional antibodies contained in the library. Here, we describe the construction of a novel, high quality, synthetic single domain antibody library dubbed Predator. The library is based on the HEL4 domain antibody with the addition of recently reported mutations concerning the amino acid composition at positions critical for the folding characteristics and aggregation propensities of domain antibodies. As a unique feature, the CDR3 of the library was designed to mimic the natural human immune response by designating amino acids known to be prevalent in functional antibodies to the diversity in CDR3. CDR randomizations were performed using trinucleotide synthesis to avoid the presence of stop codons. Furthermore a novel cycle free elongation method was used for the conversion of the synthesized single stranded DNA containing the randomized CDRs into double stranded DNA of the library. In addition a modular approach has been adopted for the scaffold in which each CDR region is flanked by unique restrictions sites, allowing easy affinity maturation of selected clones by CDR shuffling. To validate the quality of the library, one round phage display selections were performed on purified antigens and highly complex antigen mixtures such as cultured eukaryotic cells resulting in several specific binders. The further characterization of some of the selected clones, however, indicates a reduction in thermodynamic stability caused by the inclusion the additional mutations to the HEL4 scaffold.

  6. Characterization of antibody-chelator conjugates: Determination of chelator content by terbium fluorescence titration

    Energy Technology Data Exchange (ETDEWEB)

    Brandt, K.D.; Schnobrich, K.E.; Johnson, D.K. (Abbott Laboratories, Department 90M, Abbott Park, IL (United States))

    1991-01-01

    Fluorescence titrations were performed by adding varying mole ratios of terbium(III) to antibody conjugates formed by benzyl isothiocyanate derivatives of three different polyaminopolycarboxylate chelators (NTA, EDTA, and DTPA) and the results compared to values for average chelator content obtained by cobalt-57 binding assays. For two different murine monoclonal antibodies, the average chelator content obtained by terbium fluorescence titration correlated closely with that measured by the cobalt-57 binding assay. It is concluded that lanthanide fluorescence titrations provide a useful alternative to radiometal binding assays for the determination of chelator content in protein-chelator conjugates.

  7. Label free checkerboard assay to determine overlapping epitopes of Ebola virus VP-40 antibodies using surface plasmon resonance.

    Science.gov (United States)

    Anderson, George P; Liu, Jinny L; Zabetakis, Dan; Legler, Patricia M; Goldman, Ellen R

    2017-03-01

    Immunoassay formats, in which antibodies provide sensitivity and specificity, are often utilized to provide rapid and simple diagnostic tests. Surface plasmon resonance is frequently used to evaluate the suitability of antibodies by determining binding kinetics to agents or surrogate antigens. We used SPR to evaluate a number of commercial monoclonal antibodies as well as single domain antibodies produced in-house. All the antibodies targeted the Ebola virus viral protein 40 (VP40). We determined the ability of each antibody to bind to immobilized VP40, and ensured they did not bind Ebola glycoprotein or the nucleoprotein. A subset of the monoclonal antibodies was immobilized to characterize antigen capture in solution. It can be advantageous to utilize antibodies that recognize distinct epitopes when choosing reagents for detection and diagnostic assays. We determined the uniqueness of the epitope recognized by the anti-VP40 antibodies using a checkerboard format that exploits the 6×6 array of interactions monitored by the Bio-Rad ProteOn XPR36 SPR instrument. The results demonstrate the utility of surface plasmon resonance to characterize monoclonal and recombinant antibodies. Additionally, the analysis presented here enabled the identification of pairs of anti-VP40 antibodies which could potentially be utilized in sandwich type immunoassays for the detection of Ebola virus. Published by Elsevier B.V.

  8. Frequency of anti-Chlamydia trachomatis antibodies in infertile women referred to Tabriz Al-Zahra hospital

    Directory of Open Access Journals (Sweden)

    Mahtab Sattari

    2017-08-01

    Full Text Available Background: Infertility is one of the major issues in society and its incidence is estimated to be almost 10-15%. Chlamydia trachomatis (C. trachomatis is an important cause of sexually transmitted diseases leading to infertility. Objective: This study was designed to determine the frequency of anti-C. trachomatis antibodies in infertile women at Al-zahra hospital, Tabriz, Iran. Materials and Methods: In this cross-sectional study, the blood samples were collected randomly from 184 infertile women (case group and 100 pregnant women (control group. The frequency of specific IgG and IgM anti-C. trachomatis antibodies were evaluated using ELISA method. Results: The frequency of IgG anti-C. trachomatis antibody in the control and case groups was 18% and 35.88%, respectively. IgM anti-C. trachomatis antibody was found in 2% of controls and 5.44% of infertile women. Our results showed the significant differences between the case and control groups in anti-C. trachomatis antibodies (IgG, p=0.035 and IgM, p=0.004. Also, no significant relation was seen between the frequency of anti-C. trachomatis antibodies and age, location, and tubal factor infertility in our two study groups. Conclusion: According to high frequency of antibody anti-C. trachomatis among infertile women in competition to the control group, evaluation and treatment of Chlamydia infections is necessary in these patients

  9. Anti-xanthine oxidase antibodies in sera and synovial fluid of patients with rheumatoid arthritis and other joint inflammations

    International Nuclear Information System (INIS)

    Arrar, L.; Hanachi, N.; Rouba, K.; Charef, N.; Khennouf, S.; Baghiani, A.

    2008-01-01

    Objective was to study anti-bovine milk xanthine oxidoreductase XOPR antibody levels in synovial fluid as well as in serum of patients suffering from rheumatoid affections to assess a possible correlation between antibody titres and severity of disease. Sera and synovial fluids were collected from volunteer donors at Setif University Hospital, Setif, Algeria from 2001-2007 with the consent of patients. Human IgG and IgM levels of free and bound anti-bovine milk XOR antibodies were determined using bovine XOR as antigen, with enzyme-linked immunosorbent assay ELISA. Serum IgG anti-bovine milk XOR titres in 30 healthy normal subjects 2.74+-2.31 microgram/mL are in agreement with that reported in the literature. Immunoglobulin G and IgM anti-bovine milk XOR antibody titres were found to be significantly higher in serum from patients with rheumatoid arthritis RA and latex positives subjects. Synovial IgM antibody titres to bovine XOR were found to be significantly higher in rheumatoid arthritis patients compared to patients with other joint inflammations. In rheumatoid arthritis patients, high concentrations of antibodies against XOR were noticed. These antibodies may play a major role in RA by inhibiting both xanthine and NADH oxidase activities of XOR. They may also play a key role in eliminating XOR from serum and synovial fluid positive role but unfortunately, immune complex formation could also activate complement and participate in self maintenance of inflammation. (author)

  10. Localization of immunodominant epitopes within the "a" determinant of hepatitis B surface antigen using monoclonal antibodies.

    Science.gov (United States)

    Golsaz-Shirazi, Forough; Mohammadi, Hamed; Amiri, Mohammad Mehdi; Khoshnoodi, Jalal; Kardar, Gholam Ali; Jeddi-Tehrani, Mahmood; Shokri, Fazel

    2016-10-01

    The common "a" determinant is the major immunodominant region of hepatitis B surface antigen (HBsAg) shared by all serotypes and genotypes of hepatitis B virus (HBV). Antibodies against this region are thought to confer protection against HBV and are essential for viral clearance. Mutations within the "a" determinant may lead to conformational changes in this region, which can affect the binding of neutralizing antibodies. There is an increasing concern about identification and control of mutant viruses which is possible by comprehensive structural investigation of the epitopes located within this region. Anti-HBs monoclonal antibodies (mAbs) against different epitopes of HBsAg are a promising tool to meet this goal. In the present study, 19 anti-HBs mAbs were employed to map epitopes localized within the "a" determinant, using a panel of recombinant mutant HBsAgs. The topology of the epitopes was analyzed by competitive enzyme-linked immunosorbent assay (ELISA). Our results indicate that all of the mAbs seem to recognize epitopes within or in the vicinity of the "a" determinant of HBsAg. Different patterns of binding with mutant forms were observed with different mAbs. Amino acid substitutions at positions 123, 126, 129, 144, and 145 dramatically reduced the reactivity of antibodies with HBsAg. The T123N mutation had the largest impact on antibody binding to HBsAg. The reactivity pattern of our panel of mAbs with mutant forms of HBsAg could have important clinical implications for immunoscreening, diagnosis of HBV infection, design of a new generation of recombinant HB vaccines, and immunoprophylaxis of HBV infection as an alternative to therapy with hepatitis B immune globulin (HBIG).

  11. Splenectomy associated changes in IgM memory B cells in an adult spleen registry cohort.

    Directory of Open Access Journals (Sweden)

    Paul U Cameron

    Full Text Available Asplenic patients have a lifelong risk of overwhelming post-splenectomy infection and have been reported to have low numbers of peripheral blood IgM memory B cells. The clinical value of quantitation of memory B cells as an indicator of splenic abnormality or risk of infection has been unclear. To assess changes in B cell sub-populations after splenectomy we studied patients recruited to a spleen registry (n = 591. A subset of 209 adult asplenic or hyposplenic subjects, and normal controls (n = 140 were tested for IgM memory B cells. We also determined a changes in IgM memory B cells with time after splenectomy using the cross-sectional data from patients on the registry and b the kinetics of changes in haematological markers associated with splenectomy(n = 45. Total B cells in splenectomy patients did not differ from controls, but memory B cells, IgM memory B cells and switched B cells were significantly (p<0.001 reduced. The reduction was similar for different indications for splenectomy. Changes of asplenia in routine blood films including presence of Howell-Jolly bodies (HJB, occurred early (median 25 days and splenectomy associated thrombocytosis and lymphocytosis peaked by 50 days. There was a more gradual decrease in IgM memory B cells reaching a stable level within 6 months after splenectomy. IgM memory B cells as proportion of B cells was the best discriminator between splenectomized patients and normal controls and at the optimal cut-off of 4.53, showed a true positive rate of 95% and false positive rate of 20%. In a survey of 152 registry patients stratified by IgM memory B cells around this cut-off there was no association with minor infections and no registry patients experienced OPSI during the study. Despite significant changes after splenectomy, conventional measures of IgM memory cells have limited clinical utility in this population.

  12. Splenectomy Associated Changes in IgM Memory B Cells in an Adult Spleen Registry Cohort

    Science.gov (United States)

    Cameron, Paul U.; Jones, Penelope; Gorniak, Malgorzata; Dunster, Kate; Paul, Eldho; Lewin, Sharon; Woolley, Ian; Spelman, Denis

    2011-01-01

    Asplenic patients have a lifelong risk of overwhelming post-splenectomy infection and have been reported to have low numbers of peripheral blood IgM memory B cells. The clinical value of quantitation of memory B cells as an indicator of splenic abnormality or risk of infection has been unclear. To assess changes in B cell sub-populations after splenectomy we studied patients recruited to a spleen registry (n = 591). A subset of 209 adult asplenic or hyposplenic subjects, and normal controls (n = 140) were tested for IgM memory B cells. We also determined a) changes in IgM memory B cells with time after splenectomy using the cross-sectional data from patients on the registry and b) the kinetics of changes in haematological markers associated with splenectomy(n = 45). Total B cells in splenectomy patients did not differ from controls, but memory B cells, IgM memory B cells and switched B cells were significantly (psplenectomy. Changes of asplenia in routine blood films including presence of Howell-Jolly bodies (HJB), occurred early (median 25 days) and splenectomy associated thrombocytosis and lymphocytosis peaked by 50 days. There was a more gradual decrease in IgM memory B cells reaching a stable level within 6 months after splenectomy. IgM memory B cells as proportion of B cells was the best discriminator between splenectomized patients and normal controls and at the optimal cut-off of 4.53, showed a true positive rate of 95% and false positive rate of 20%. In a survey of 152 registry patients stratified by IgM memory B cells around this cut-off there was no association with minor infections and no registry patients experienced OPSI during the study. Despite significant changes after splenectomy, conventional measures of IgM memory cells have limited clinical utility in this population. PMID:21829713

  13. The etiology of Rubella IgM positivity in patients with rubella-like illness in Iran from 2011 to 2013.

    Science.gov (United States)

    Khorrami, Seyed Mahmood Seyed; Mokhtari-Azad, Talat; Yavarian, Jila; Nasab, Gazal Sadat Fatemi; Naseri, Maryam; Jandaghi, Nazanin Zahra Shafiei

    2015-11-01

    Rubella is a mild self-limiting contagious viral disease caused by the rubella virus (RV). Although symptoms are often mild, the concern is centralized around the possible effect on a fetus growth and development in case of primary infection during early months of pregnancy. Recently acquired rubella is commonly confirmed by RV-specific IgM antibody detection in the serum. However, rubella primary infection is not always the only cause of IgM positivity. Other possible causes of rubella IgM positivity may include IgM persistence following vaccination or naturally acquired infection or even re-infection. Moreover, nonspecific IgM reactivity can cause false-positive results. There are few articles to differentiate the aetiology of rash in rubella-like illnesses. However, limited studies have been conducted on clarifying the source of IgM positivity in these cases. This article reports the study of 10,896 clinical cases demonstrating rubella-like illness between 2011 and 2013 in Iran. The rate of IgM positivity among these cases was 0.52% (57 cases). As predicted based on the high coverage of vaccination in Iran fewer than 16% of cases with ELISA IgM positive result, were due to current rubella primary infections. The greater part of the positive IgM reactions occurred in cross reactivity with other viruses (31.6%) or in prolonged IgM response post vaccination (24.6%). This research confirmed that the positive result of rubella IgM assay in vaccinated individuals is mainly caused by prolonged IgM production, rubella re-infection, and false positivity due to infection with other viruses, rather than the rubella primary infection itself. © 2015 Wiley Periodicals, Inc.

  14. The effect of X-rays on the precursors of antibody forming cells (B cells) as measured with the in vitro limiting dilution assay

    NARCIS (Netherlands)

    Lubbe, F.H.; Hooijkaas, H.; Preesman, A.A.

    1982-01-01

    The effect of X-irradiation upon murine antibody-forming cell precursors (B cells) was established in cultures of spleen cells stimulated with the B cell mitogen lipopolysaccharide (LPS). At day 5 and 7 the numbers of IgM- and IgG2-secreting cells were determined in cultures of irradiated and

  15. Detection and differentiation of cytomegalovirus antibodies by radioimmunoassay

    International Nuclear Information System (INIS)

    Jankowski, M.A.; Gut, W.; Nawrocka, E.; Kantoch, M.

    1980-01-01

    A sensitive radioimmunological method was adapted for the assay of anticytomegalovirus IgG and IgM antibodies. Binding of immunoglobulins G with cytomegalovirus (CMV) antigens was greatly reduced when an antigen prepared by sonication of the infected cells in glycine buffer was used in place of the antigen obtained by freezing and thawing of the cells. The application of labelled serum against the Fc fragment of IgG made it possible to identify selectively the antibodies bound with the virus antigen by antigenic determinants. (Auth.)

  16. Fundamental roles of the innate-like repertoire of natural antibodies in immune homeostasis

    Directory of Open Access Journals (Sweden)

    Jaya eVas

    2013-02-01

    Full Text Available The composition of the early immune repertoire is biased with prominent expression of spontaneously arising B-cell clones that produce IgM with recurrent and often autoreactive binding specificities. Amongst these naturally-arising antibodies (NAbs are IgM antibodies that specifically recognize damaged and senescent cells, often via oxidation-associated neo-determinants. These NAbs are present from birth and can be further boosted by apoptotic cell challenge. Recent studies have shown that IgM NAb to apoptotic cells can enhance phagocytic clearance, as well as suppress pro-inflammatory responses induced via Toll-like receptors, and block pathogenic IgG-immune complex (IC-mediated inflammatory responses. Specific antibody effector functions appear to be involved, as these anti-inflammatory properties are dependent on IgM-mediated recruitment of the early recognition factors of complement. Clinical surveys have suggested that anti-AC IgM NAbs may modulate disease activity in some patients with autoimmune disease. In mechanistic studies, anti-AC NAbs were shown to act in dendritic cells by inhibition of the Mitogen Activated Protein Kinase (MAPK pathway, a primary signal transduction pathway that controls inflammatory responses. This immunomodulatory pathway has an absolute requirement for the induction of MAPK Phosphatase-1. Taken together, recent studies have elucidated the novel properties of a class of protective NAbs, which may directly blunt inflammatory responses through a primitive pathway for regulation of the innate immune system.

  17. Complement deposition in autoimmune hemolytic anemia is a footprint for difficult-to-detect IgM autoantibodies

    NARCIS (Netherlands)

    Meulenbroek, Elisabeth M.; de Haas, Masja; Brouwer, Conny; Folman, Claudia; Zeerleder, Sacha S.; Wouters, Diana

    2015-01-01

    In autoimmune hemolytic anemia autoantibodies against erythrocytes lead to increased clearance of the erythrocytes, which in turn results in a potentially fatal hemolytic anemia. Depending on whether IgG or IgM antibodies are involved, response to therapy is different. Proper identification of the

  18. A mouse monoclonal antibody against a polymorphic determinant in a defined subset of DR molecules.

    Science.gov (United States)

    Mazzilli, M C; Tanigaki, N; Cascino, I; Costanzi Porrini, S; Trabace, S; Cappellacci, S; Testa, L; Gandini, E

    1986-06-01

    The hybridoma technique was used to produce a mouse monoclonal antibody, designated as XI 21.4, which belongs to the IgG2a class. It is active in complement-dependent cytotoxicity and detects a B-cell antigenic determinant associated with DR1, DR4, DRw10, and, possibly, DRw9. Microfingerprinting of the immunoprecipitate from a homozygous DR4 cell line shows a typical alpha DR pattern and a beta pattern coinciding with that of DR4 molecules.

  19. Determination of neuronal antibodies in suspected and definite Creutzfeldt-Jakob disease

    OpenAIRE

    Grau-Rivera, O.; Sánchez del Valle Díaz, Raquel; Saiz Hinajeros, Albert; Molinuevo, José L.; Bernabé, Reyes; Munteis, Elvira; Pujadas, Francesc; Salvador, Antoni; Saura, Júlia; Ugarte, Antonio; Titulaer, Maarten; Dalmau Obrador, Josep; Graus Ribas, Francesc

    2014-01-01

    IMPORTANCE Creutzfeldt-Jakob disease (CJD) and autoimmune encephalitis with antibodies against neuronal surface antigens (NSA-abs) may present with similar clinical features. Establishing the correct diagnosis has practical implications in the management of care for these patients. OBJECTIVE To determine the frequency of NSA-abs in the cerebrospinal fluid of patients with suspected CJD and in patients with pathologically confirmed (ie, definite) CJD. DESIGN, SETTING, AND PARTICIPANTS A mixed ...

  20. Prevalence and basic determinants of hepatitis C antibodies in medical students in Katowice, Poland.

    Science.gov (United States)

    Braczkowska, Bogumiła; Kowalskan, Małgorzata; Zejda, Jan E; Beniowski, Marek; Kulawik, Ilona; Kuleszyński, Jaromir; Jagodziński, Leszek

    2006-01-01

    Potential exposure of medical students to infectious material and the lack of specific prevention measures could contribute to an increased risk of HCV infection in this group. To find out if the occurrence of HCV antibodies is more frequent in medical students than in non-medical students and to determine risk factors for HCV antibodies in students. A cross-sectional study performed in 2003-2004 included 566 medical students (Medical University, Katowice) and 517 students of non-medical faculties (University, Katowice). HCV antibodies were determined using the 3rd generation EIA test, in doubtful cases a Western-blot INNO-LIA-HCV confirmation test was applied. Risk factors were identified based on questionnaire derived data. Between-group difference in the prevalence of positive anti-HCV tests was evaluated by means of ch2 test and candidates for risk factors were evaluated according to logistic odds ratios. Prevalence of HCV antibodies was statistically insignificantly lower in medical students (1.4%) than that in non-medical students (1.9%). In the group of 1068 subjects with no history of ever-diagnosed viral hepatitis the respective figures were 1.4% and 1.7%. In this group the presence of the positive anti-HCV tests was associated with dermal cosmetic interventions in the past (logOR=4.85), percutaneous medical interventions (excluding surgery) (logOR=3.18) and individual history of blood transfusion (logOR=1.71). However, the results were not statistically significant. Medical students at the Medical University in Katowice, Poland are not at increased risk of HCV infection. The findings suggest an important role of cosmetic and percutaneous medical interventions, and blood transfusion as risk factors for HCV infection.

  1. Pediatric Selective IgM Immunodeficiency

    Directory of Open Access Journals (Sweden)

    Marc F. Goldstein

    2008-01-01

    Full Text Available Objective. Limited information exists on features of pediatric Selective IgM immunodeficiency (SIgMID. Previously published pediatric cases and 2 new cases are reviewed. Methods. English literature from PubMed and references from relevant articles were reviewed. Previously reported cases and 2 new cases from an allergy/immunology practice were analyzed. Results. Forty-nine reported cases of SIgMID presented with respiratory infections (77.6%, gastrointestinal disease (16.3%, skin disease (12.2%, and meningitis (8.2%. Mean serum IgM level was 16.5±13.8 mg/dL. Two patients were identified with SIgMID among 6300 active pediatric patients (0.03% presenting with asthma, vasomotor rhinitis, and recurrent respiratory infections. In the 51 cases reported, none developed lymphoproliferative disease nor evolved into panhypogammaglobulinemia; four fatalities were reported. Conclusions. The prevalence of SIgMID in our pediatric population was 0.03%. In general, respiratory infections are the common comorbid conditions. Death and autoimmune disease are uncommon complications of pediatric SIgMID.

  2. Comparison of Rubazyme-M and MACRIA for the detection of rubella-specific IgM

    Energy Technology Data Exchange (ETDEWEB)

    Best, J.M.; Palmer, S.J.; Morgan-Capner, P.; Hodgson, J. (Saint Thomas' Hospital, London (UK))

    1984-02-01

    One hundred and eighty-six carefully selected sera were tested for rubella-specific IgM by Rubazyme-M (Abbott Diagnostics) and an M-antibody capture radioimmunoassay (MACRIA). Eleven of these sera were from cases of infectious mononucleosis, six of which gave positive results in MACRIA, while one gave a positive result in Rubazyme-M. Of the remaining 175 sera, 158 gave concordant results whilst 17 sera gave discordant results; these 17 were also tested by serum fractionation. Problems were encountered with all assay systems used. It is therefore recommended that the results of all tests for rubella-specific IgM should be interpreted with caution.

  3. IgM monoclonal gammopathy and neuropathy in two siblings

    DEFF Research Database (Denmark)

    Jensen, T S; Schrøder, H D; Jønsson, V

    1988-01-01

    patients contained antibodies directed to bovine peripheral nerve myelin as determined by ELISA technique and to normal human peripheral nerve myelin as demonstrated by indirect immunofluorescence histochemistry. These siblings may have a genetic predisposition to the formation of autoantibodies...

  4. Sensitive double-antibody method for simultaneous determination of insulin and growth hormone

    International Nuclear Information System (INIS)

    Koparanova, O.; Sotirov, G.; Tyrkolev, N.

    1982-01-01

    A method is described for simultaneous determination of insulin and growth hormone in one sample, using double-antibody technique. The method is characterized by appreciable sensitivity (2.5 μE/ml for insulin and a.2 ng/ml for growth hormone), exactness (variation quotient 6-16 per cent) and reproducibility (96.9-117 per cent). There was no statistically significant difference in the insulin and growth hormone values of the same sera, determined by the here suggested and the standard methods. The necessary test material for examination of either hormone is minimal (0.2 ml). One may thus extend the possibilities for radioimmunologic determination of insulin and growth hormone, when only minor amounts of serum or other biological fluid are available. The method is also less time consuming. Results are reported of statistical processing of an experimental model and different sera determined by the standard method and the one described by the authors. (author)

  5. Seroprevalence of anti-Chlamydia trachomatis IgM in neonatal respiratory tract infections in Hungary.

    Science.gov (United States)

    Balla, Eszter; Donders, Gilbert G G; Petrovay, Fruzsina; Urbán, Edit

    2017-08-04

    To determine the seroprevalence of specific IgM indicative of respiratory tract infection (RTI) due to Chlamydia trachomatis (CT) among symptomatic infants. A descriptive study was conducted on young infants up to 5 months old at the Bacterial Sexually Transmitted Infections Reference Laboratory, National Centre for Epidemiology, Budapest, covering the period 2008-2016. Serum samples from infants suffering from RTIs were screened with a micro-immunofluorescence test (Focus, Cypress, USA) for the presence of anti-Chlamydia trachomatis-specific IgM. A parallel Bordetella pertussis screening was performed by an indirect immunofluorescence test (Euroimmun, Lübeck, Germany) that detected specific IgM. The CT-specific serum IgM was highly reactive in 50 (19.1 %) of the 262 neonates with RTIs, while all proved negative for Bordetella pertussis-specific IgM. Vertically transmitted C. trachomatis must be regarded as a common pathogen among symptomatic neonates with RTIs in Hungary. Routine screening and treatment of pregnant women could be one option to help prevent these conditions. Focused laboratory testing based on raised clinical awareness should enable early diagnosis and appropriate therapy for symptomatic infants.

  6. Truly selective primary IgM deficiency is probably very rare.

    Science.gov (United States)

    Janssen, L M A; Macken, T; Creemers, M C W; Pruijt, J F M; Eijk, J J J; de Vries, E

    2018-02-01

    Isolated decreased serum-immunoglobulin (Ig)M has been associated with severe and/or recurrent infections, atopy and autoimmunity. However, the reported high prevalence of clinical problems in IgM-deficient patients may reflect the skewed tertiary centre population studied so far. Also, many papers on IgM deficiency have included patients with more abnormalities than simply IgM-deficiency. We studied truly selective primary IgM deficiency according to the diagnostic criteria of the European Society for Immunodeficiencies (ESID) (true sIgMdef) by reviewing the literature (261 patients with primary decreased serum-IgM in 46 papers) and analysing retrospectively all patients with decreased serum-IgM in a large teaching hospital in 's-Hertogenbosch, the Netherlands [1 July 2005-23 March 2016; n = 8049 IgM IgA normal/IgM deficiencies (unPAD) instead. The diagnosis was often uncertain (possible sIgMdef): data on IgG subclasses and/or vaccination responses were lacking in 192 of 261 (74%) literature cases and 42 of 45 (93%) cases from our cohort. Our results also illustrate the clinical challenge of determining the relevance of a serum sample with decreased IgM; a larger cohort of true sIgMdef patients is needed to explore fully its clinical consequences. The ESID online Registry would be a useful tool for this. © 2017 British Society for Immunology.

  7. Drug-to-antibody determination for an antibody-drug-conjugate utilizing cathepsin B digestion coupled with reversed-phase high-pressure liquid chromatography analysis.

    Science.gov (United States)

    Adamo, Michael; Sun, Guoyong; Qiu, Difei; Valente, Joseph; Lan, Wenkui; Song, Hangtian; Bolgar, Mark; Katiyar, Amit; Krishnamurthy, Girija

    2017-01-20

    Antibody drug conjugates or ADCs are currently being evaluated for their effectiveness as targeted chemotherapeutic agents across the pharmaceutical industry. Due to the complexity arising from the choice of antibody, drug and linker; analytical methods for release and stability testing are required to provide a detailed understanding of both the antibody and the drug during manufacturing and storage. The ADC analyzed in this work consists of a tubulysin drug analogue that is randomly conjugated to lysine residues in a human IgG1 antibody. The drug is attached to the lysine residue through a peptidic, hydrolytically stable, cathepsin B cleavable linker. The random lysine conjugation produces a heterogeneous mixture of conjugated species with a variable drug-to-antibody ratio (DAR), therefore, the average amount of drug attached to the antibody is a critical parameter that needs to be monitored. In this work we have developed a universal method for determining DAR in ADCs that employ a cathepsin B cleavable linker. The ADC is first cleaved at the hinge region and then mildly reduced prior to treatment with the cathepsin B enzyme to release the drug from the antibody fragments. This pre-treatment allows the cathepsin B enzyme unrestricted access to the cleavage sites and ensures optimal conditions for the cathepsin B to cleave all the drug from the ADC molecule. The cleaved drug is then separated from the protein components by reversed phase high performance liquid chromatography (RP-HPLC) and quantitated using UV absorbance. This method affords superior cleavage efficiency to other methods that only employ a cathepsin digestion step as confirmed by mass spectrometry analysis. This method was shown to be accurate and precise for the quantitation of the DAR for two different random lysine conjugated ADC molecules. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Complement deposition in autoimmune hemolytic anemia is a footprint for difficult-to-detect IgM autoantibodies

    Science.gov (United States)

    Meulenbroek, Elisabeth M.; de Haas, Masja; Brouwer, Conny; Folman, Claudia; Zeerleder, Sacha S.; Wouters, Diana

    2015-01-01

    In autoimmune hemolytic anemia autoantibodies against erythrocytes lead to increased clearance of the erythrocytes, which in turn results in a potentially fatal hemolytic anemia. Depending on whether IgG or IgM antibodies are involved, response to therapy is different. Proper identification of the isotype of the anti-erythrocyte autoantibodies is, therefore, crucial. However, detection of IgM autoantibodies can be challenging. We, therefore, set out to improve the detection of anti-erythrocyte IgM. Direct detection using a flow cytometry-based approach did not yield satisfactory improvements. Next, we analyzed whether the presence of complement C3 on a patient’s erythrocytes could be used for indirect detection of anti-erythrocyte IgM. To this end, we fractionated patients’ sera by size exclusion chromatography and tested which fractions yielded complement deposition on erythrocytes. Strikingly, we found that all patients with C3 on their erythrocytes according to standard diagnostic tests had an IgM anti-erythrocyte component that could activate complement, even if no such autoantibody had been detected with any other test. This also included all tested patients with only IgG and C3 on their erythrocytes, who would previously have been classified as having an IgG-only mediated autoimmune hemolytic anemia. Depleting patients’ sera of either IgG or IgM and testing the remaining complement activation confirmed this result. In conclusion, complement activation in autoimmune hemolytic anemia is mostly IgM-mediated and the presence of covalent C3 on patients’ erythrocytes can be taken as a footprint of the presence of anti-erythrocyte IgM. Based on this finding, we propose a diagnostic workflow that will aid in choosing the optimal treatment strategy. PMID:26354757

  9. Clinical evaluation of dengue RNA, NS1, and IgM for diagnosis of dengue in Southern China.

    Science.gov (United States)

    Chen, Xinliang; Chen, Rui; Gu, Wenshen; He, Jian; Cai, Weipeng; Li, Jiajia; Duan, Chaohui; Yan, Haiyan

    2016-01-01

    In 2014, a large outbreak of dengue occurred in Guangzhou, China. This outbreak prompted us to evaluate NS1 and RNA for the early diagnosis of acute dengue infection, in addition to the combination with IgM antibody. We aimed to find the differences of three assays about dengue diagnosis. This study was an evaluation of diagnosis test. Based on WHO criteria 2009, dengue RNA, NS1, and IgM/IgG were detected from 294 patients (180 dengue patients, 114 non-dengue patients) by three diagnostic kits made in China. The χ(2) test, sensitivity, and specificity were used in statistical analysis. The ratios of dengue patients with low platelet counts (dengue patients (P Dengue NS1 was shown sensitive (93.9%) for diagnostic use. RNA had a better performance with 98.1% of sensitivity from day 1 to day 4 after illness onset. IgM performed better at day 5 or more with 74.0% of sensitivity. The diagnostic rate using a combination of RNA and IgM was 97.8% and 96.7% using NS1 and IgM. A patient with low platelet and white blood cell counts needs additional tests for dengue during an epidemic. RNA and NS1 were most valuable for early diagnosis of dengue, whereas IgM was best suited as a supplementary method for patients at day 5 or more after illness onset. © 2015 Wiley Periodicals, Inc.

  10. Generation and characterization of rabbit polyclonal antibodies against Vasohibin-2 for determination of its intracellular localization.

    Science.gov (United States)

    Sun, Jie; Tu, Min; Han, Bei; Xue, Xiaofeng; Zhang, Ye; Wei, Jishu; Chen, Jianmin; Lu, Zipeng; An, Yong; Cai, Baobao; Lv, Nan; Jiang, Kuirong; Miao, Yi; Gao, Wentao

    2013-07-01

    Vasohibin-2 was recently identified as an important pro-angiogenesis factor in solid tumor and intracellular localization of its variants is important for elucidating the downstream mechanism(s) of its effects. Currently there are no reported antibodies affordable for intracellular localization. The aim of this study was to generate and characterize polyclonal antibodies against Vasohibin-2 and to determine the intracellular localization of Vasohibin-2. In this study, two polypeptides were synthesized and one prokaryotic Vasohibin-2 recombinant protein was custom-made. New Zealand rabbits were immunized with the polypeptide mixture and prokaryotic recombinant protein, respectively. The purified antibodies from the antiserum were validated by ELISA, western blotting (WB), immunofluorescence (IF), immunohistochemistry (IHC) and immunoprecipitation (IP). In order to determine intracellular localization, the cytoplasmic and nuclear proteins of the human liver cancer cell line HepG2 were isolated for the detection of Vasohibin-2 by western blotting. Vasohibin-2 cDNA, coding for 311 and 355 amino acid residues, fused with or without a DDK/V5 tag at the c-terminus, respectively, was cloned into the Lv-CMV-EGFP vector. Lentiviruses were successfully packaged. Vasohibin-2-overexpressing HepG2-VASH2 (355 amino acid residues) and HepG2-VASH2-V5 (311 amino acid residues fused with V5 tag at the c-terminus) human liver cancer cell lines were established. Approximately 1-2x106 HepG2, HepG2-VASH2 and HepG2-VASH2-V5 cells were injected subcutaneously into the flanks of BALB/c nude mice. Xenograft tumors were harvested for immunohistochemistry. HepG2 cells were transiently transfected with the Lv-CMV-EGFP vectors containing Vasohibin-2 cDNA (coding for 311/355 amino acid residues with a DDK tag at the c-terminal), followed by anti-DDK immunofluorescence. The antibodies obtained were able to detect human VASH2 successfully as applied in western blotting, IF, IHC and IP. Results from

  11. Prevalence of serum antibodies to TORCH infection in the first trimester of the pregnancy in Kashan, Iran

    Directory of Open Access Journals (Sweden)

    sareh bagheri josheghani

    2015-03-01

    Full Text Available Introduction:TORCH infections causing via Toxoplasma gondii, other microorganisms (e.g., Treponema pallidum, Rubella virus, Cytomegalovirus (CMV and the Herpes Simplex Virus (HSV types 1 and 2 during the first trimester of pregnancy can lead to severe fetal anomalies or even fetal loss. The current study determined the serological data of TORCH infections in women who were in their first trimesters of pregnancy.This descriptive study was carried out on 80 pregnant women in their first trimester in Kashan, Iran. Methods: To detect specific IgM antibodies and specific IgG antibodies against the TORCH infections via ELISA, Sera were collected from the pregnant women. Results: The specific IgG antibodies were found to be positive in 30(37.5% cases for toxoplasmosis, in 74 (92.5% cases for the Rubella virus, in 79(98.8% cases for CMV and in 73 samples (91.3% for the HSV types 1 and 2 infection. 3.8% of cases were found to be seropositive for Toxoplasma IgM antibody (95% CI, 0.38-7.9, 5% were positive for CMV IgM antibody (95% CI, 0.23-9.77 and 7.5% were positive for the HSV IgM antibody (95% CI, 1.8-13.2. 63.8% of pregnant women were at risk for at least to one of the TORCH agents. Conclusion: This study showed a high prevalence of infections caused by TORCH agents among pregnant women. Therefore, national screening programmed is necessary to screen the TORCH infections routinely and to prevent and treat congenital TORCH infection

  12. Prevalence of serum antibodies to TORCH infection in the first trimester of the pregnancy in Kashan, Iran

    Directory of Open Access Journals (Sweden)

    sareh bagheri josheghani

    2015-03-01

    Full Text Available Introduction:TORCH infections causing via Toxoplasma gondii, other microorganisms (e.g., Treponema pallidum, Rubella virus, Cytomegalovirus (CMV and the Herpes Simplex Virus (HSV types 1 and 2 during the first trimester of pregnancy can lead to severe fetal anomalies or even fetal loss. The current study determined the serological data of TORCH infections in women who were in their first trimesters of pregnancy.This descriptive study was carried out on 80 pregnant women in their first trimester in Kashan, Iran.Methods: To detect specific IgM antibodies and specific IgG antibodies against the TORCH infections via ELISA, Sera were collected from the pregnant women.Results: The specific IgG antibodies were found to be positive in 30(37.5% cases for toxoplasmosis, in 74 (92.5% cases for the Rubella virus, in 79(98.8% cases for CMV and in 73 samples (91.3% for the HSV types 1 and 2 infection. 3.8% of cases were found to be seropositive for Toxoplasma IgM antibody (95% CI, 0.38-7.9, 5% were positive for CMV IgM antibody (95% CI, 0.23-9.77 and 7.5% were positive for the HSV IgM antibody (95% CI, 1.8-13.2. 63.8% of pregnant women were at risk for at least to one of the TORCH agents.Conclusion: This study showed a high prevalence of infections caused by TORCH agents among pregnant women. Therefore, national screening programmed is necessary to screen the TORCH infections routinely and to prevent and treat congenital TORCH infection

  13. Genetic variants are major determinants of CSF antibody levels in multiple sclerosis.

    Science.gov (United States)

    Goris, An; Pauwels, Ine; Gustavsen, Marte W; van Son, Brechtje; Hilven, Kelly; Bos, Steffan D; Celius, Elisabeth Gulowsen; Berg-Hansen, Pål; Aarseth, Jan; Myhr, Kjell-Morten; D'Alfonso, Sandra; Barizzone, Nadia; Leone, Maurizio A; Martinelli Boneschi, Filippo; Sorosina, Melissa; Liberatore, Giuseppe; Kockum, Ingrid; Olsson, Tomas; Hillert, Jan; Alfredsson, Lars; Bedri, Sahl Khalid; Hemmer, Bernhard; Buck, Dorothea; Berthele, Achim; Knier, Benjamin; Biberacher, Viola; van Pesch, Vincent; Sindic, Christian; Bang Oturai, Annette; Søndergaard, Helle Bach; Sellebjerg, Finn; Jensen, Poul Erik H; Comabella, Manuel; Montalban, Xavier; Pérez-Boza, Jennifer; Malhotra, Sunny; Lechner-Scott, Jeannette; Broadley, Simon; Slee, Mark; Taylor, Bruce; Kermode, Allan G; Gourraud, Pierre-Antoine; Sawcer, Stephen J; Andreassen, Bettina Kullle; Dubois, Bénédicte; Harbo, Hanne F

    2015-03-01

    as female gender, age at onset and severity. This is the largest study population so far investigated for the genetic influence on antibody levels in the cerebrospinal fluid in multiple sclerosis, including 6950 patients. We confirm that genetic factors underlie these antibody levels and identify both the major histocompatibility complex and immunoglobulin heavy chain region as major determinants. © The Author (2015). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  14. Genetic variants are major determinants of CSF antibody levels in multiple sclerosis

    Science.gov (United States)

    Pauwels, Ine; Gustavsen, Marte W.; van Son, Brechtje; Hilven, Kelly; Bos, Steffan D.; Celius, Elisabeth Gulowsen; Berg-Hansen, Pål; Aarseth, Jan; Myhr, Kjell-Morten; D’Alfonso, Sandra; Barizzone, Nadia; Leone, Maurizio A.; Martinelli Boneschi, Filippo; Sorosina, Melissa; Liberatore, Giuseppe; Kockum, Ingrid; Olsson, Tomas; Hillert, Jan; Alfredsson, Lars; Bedri, Sahl Khalid; Hemmer, Bernhard; Buck, Dorothea; Berthele, Achim; Knier, Benjamin; Biberacher, Viola; van Pesch, Vincent; Sindic, Christian; Bang Oturai, Annette; Søndergaard, Helle Bach; Sellebjerg, Finn; Jensen, Poul Erik H.; Comabella, Manuel; Montalban, Xavier; Pérez-Boza, Jennifer; Malhotra, Sunny; Lechner-Scott, Jeannette; Broadley, Simon; Slee, Mark; Taylor, Bruce; Kermode, Allan G.; Gourraud, Pierre-Antoine; Sawcer, Stephen J.; Andreassen, Bettina Kullle; Dubois, Bénédicte; Harbo, Hanne F.

    2015-01-01

    as female gender, age at onset and severity. This is the largest study population so far investigated for the genetic influence on antibody levels in the cerebrospinal fluid in multiple sclerosis, including 6950 patients. We confirm that genetic factors underlie these antibody levels and identify both the major histocompatibility complex and immunoglobulin heavy chain region as major determinants. PMID:25616667

  15. Antibody complementarity-determining regions (CDRs: a bridge between adaptive and innate immunity.

    Directory of Open Access Journals (Sweden)

    Elena Gabrielli

    2009-12-01

    Full Text Available It has been documented that, independently from the specificity of the native antibody (Ab for a given antigen (Ag, complementarity determining regions (CDR-related peptides may display differential antimicrobial, antiviral and antitumor activities.In this study we demonstrate that a synthetic peptide with sequence identical to V(HCDR3 of a mouse monoclonal Ab (mAb specific for difucosyl human blood group A is easily taken up by macrophages with subsequent stimulation of: i proinflammatory cytokine production; ii PI3K-Akt pathway and iii TLR-4 expression. Significantly, V(HCDR3 exerts therapeutic effect against systemic candidiasis without possessing direct candidacidal properties.These results open a new scenario about the possibility that, beyond the half life of immunoglobulins, Ab fragments may effectively influence the antiinfective cellular immune response in a way reminiscent of regulatory peptides of innate immunity.

  16. Clonal analysis of a human antimouse antibody (HAMA) response.

    Science.gov (United States)

    Thorpe, S J; Turner, C; Heath, A; Feavers, I; Vatn, I; Natvig, J B; Thompson, K M

    2003-01-01

    Circulating human antimouse antibodies (HAMAs) directed to mouse immunoglobulin G (IgG) are clinically significant, compromising mouse antibody therapy and imaging, and interfering in immunological assays. To investigate the HAMA response, 20 stable cell lines secreting human monoclonal antibodies reactive with mouse IgG were established from a donor with a history of exposure to mice. Their subclass and domain specificities were established by solid-phase binding, indirect haemagglutination assays and immunoblotting, using Igs of known subclass and Ig fragments. The heavy-chain variable region gene usage was determined for 12 HAMAs. Eight HAMAs were IgM, 11 HAMAs were IgG4 and one HAMA was IgG1, indicating an IgG4-dominated response. All of the IgG HAMAs reacted with epitopes present on the Fc portion; one was subclass-specific, nine were subclass-restricted and two were pan-IgG-reactive. Measurement of their affinities gave dissociation constants typically in the nanomolar range. Seven and five HAMAs were derived from variable heavy-chain 3 (VH3) and VH1 gene segments, respectively. The IgG HAMAs used different VH segments to the IgM HAMAs. JH regions were coded by JH4 in eight HAMAs. DH segment usage appeared to be restricted in the IgM HAMAs. Two IgG HAMAs were clonally related. These monoclonal HAMAs are potentially useful as reagents for detecting mouse IgG and as reference reagents for the investigation of the HAMA response in patients undergoing mouse monoclonal antibody therapy and for the investigation of the influence of HAMAs on immunodiagnostic tests.

  17. Improvement in Diagnosis of Histoplasma Meningitis by Combined Testing for Histoplasma Antigen and Immunoglobulin G and Immunoglobulin M Anti-Histoplasma Antibody in Cerebrospinal Fluid

    Science.gov (United States)

    Bloch, Karen C; Myint, Thein; Raymond-Guillen, Luke; Hage, Chadi A; Davis, Thomas E; Wright, Patty W; Chow, Felicia C; Woc-Colburn, Laila; Khairy, Raed N; Street, Alan C; Yamamoto, Tomotaka; Albers, Amanda; Wheat, L Joseph

    2018-01-01

    Abstract Background Central nervous system (CNS) histoplasmosis is a life-threatening condition and represents a diagnostic and therapeutic challenge. Isolation of Histoplasma capsulatum from cerebrospinal fluid (CSF) or brain tissue is diagnostic; however, culture is insensitive and slow growth may result in significant treatment delay. We performed a retrospective multicenter study to evaluate the sensitivity and specificity of a new anti-Histoplasma antibody enzyme immunoassay (EIA) for the detection of IgG and IgM antibody in the CSF for diagnosis of CNS histoplasmosis, the primary objective of the study. The secondary objective was to determine the effect of improvements in the Histoplasma galactomannan antigen detection EIA on the diagnosis of Histoplasma meningitis. Methods Residual CSF specimens from patients with Histoplasma meningitis and controls were tested for Histoplasma antigen and anti-Histoplasma immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody using assays developed at MiraVista Diagnostics. Results A total of 50 cases and 157 controls were evaluated. Fifty percent of patients with CNS histoplasmosis were immunocompromised, 14% had other medical conditions, and 36% were healthy. Histoplasma antigen was detected in CSF in 78% of cases and the specificity was 97%. Anti-Histoplasma IgG or IgM antibody was detected in 82% of cases and the specificity was 93%. The sensitivity of detection of antibody by currently available serologic testing including immunodiffusion and complement fixation was 51% and the specificity was 96%. Testing for both CSF antigen and antibody by EIA was the most sensitive approach, detecting 98% of cases. Conclusions Testing CSF for anti-Histoplasma IgG and IgM antibody complements antigen detection and improves the sensitivity for diagnosis of Histoplasma meningitis. PMID:29020213

  18. Generation, characterization and in vivo biological activity of two distinct monoclonal anti-PEG IgMs

    Energy Technology Data Exchange (ETDEWEB)

    Hashimoto, Yosuke; Shimizu, Taro; Mima, Yu [Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, 1-78-1, Sho-machi, Tokushima 770-8505 (Japan); Abu Lila, Amr S. [Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, 1-78-1, Sho-machi, Tokushima 770-8505 (Japan); Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Zagazig University, Sharqia Governorate, Zagazig 44519 (Egypt); Ishida, Tatsuhiro, E-mail: ishida@tokushima-u.ac.jp [Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, 1-78-1, Sho-machi, Tokushima 770-8505 (Japan); Kiwada, Hiroshi [Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, 1-78-1, Sho-machi, Tokushima 770-8505 (Japan)

    2014-05-15

    PEGylation, the attachment of polyethylene glycol (PEG) to nanocarriers and proteins, is a widely accepted approach to improving the in vivo efficacy of the non-PEGylated products. However, both PEGylated liposomes and PEGylated proteins reportedly trigger the production of specific antibodies, mainly IgM, against the PEG moiety, which possibly leads to a reduction in safety and therapeutic efficacy of the PEGylated products. In the present study, two monoclonal anti-PEG IgMs — HIK-M09 via immunization with an intravenous injection of PEGylated liposomes (SLs) and HIK-M11 via immunization with a subcutaneous administration of PEGylated ovalbumin (PEG-OVA) were successfully generated. The generated IgMs showed efficient reactivity to mPEG{sub 2000} conjugated to 1,2-distearoyl-sn-glycero-3-phospho-ethanolamine (DSPE), PEGylated liposome (SL) and PEG-OVA. It appears that HIK-M09 recognizes ethoxy (OCH{sub 2}CH{sub 2}) repeat units along with a terminal motif of PEG, while HIK-M11 recognizes only ethoxy repeat units of PEG. Such unique properties allow HIK-M09 to bind with dense PEG. In addition, their impact on the in vivo clearance of the PEGylated products was investigated. It was found that the generated ant-PEG IgMs induced a clearance of SL as they were intravenously administered with SL. Interestingly, the HIK-M11, generated by PEG-OVA, induced the clearance of both SL and PEG-OVA, while the HIK-M09, generated by SL, induced the clearance of SL only. We here revealed that the presence of serum anti-PEG IgM and the subsequent binding of anti-PEG IgM to the PEGylated products are not necessarily related to the enhanced clearance of the products. It appears that subsequent complement activation following anti-PEG IgM binding is the most important step in dictating the in vivo fate of PEGylated products. This study may have implications for the design, development and clinical application of PEGylated products and therapeutics. - Highlights: • Two monoclonal

  19. Generation, characterization and in vivo biological activity of two distinct monoclonal anti-PEG IgMs

    International Nuclear Information System (INIS)

    Hashimoto, Yosuke; Shimizu, Taro; Mima, Yu; Abu Lila, Amr S.; Ishida, Tatsuhiro; Kiwada, Hiroshi

    2014-01-01

    PEGylation, the attachment of polyethylene glycol (PEG) to nanocarriers and proteins, is a widely accepted approach to improving the in vivo efficacy of the non-PEGylated products. However, both PEGylated liposomes and PEGylated proteins reportedly trigger the production of specific antibodies, mainly IgM, against the PEG moiety, which possibly leads to a reduction in safety and therapeutic efficacy of the PEGylated products. In the present study, two monoclonal anti-PEG IgMs — HIK-M09 via immunization with an intravenous injection of PEGylated liposomes (SLs) and HIK-M11 via immunization with a subcutaneous administration of PEGylated ovalbumin (PEG-OVA) were successfully generated. The generated IgMs showed efficient reactivity to mPEG 2000 conjugated to 1,2-distearoyl-sn-glycero-3-phospho-ethanolamine (DSPE), PEGylated liposome (SL) and PEG-OVA. It appears that HIK-M09 recognizes ethoxy (OCH 2 CH 2 ) repeat units along with a terminal motif of PEG, while HIK-M11 recognizes only ethoxy repeat units of PEG. Such unique properties allow HIK-M09 to bind with dense PEG. In addition, their impact on the in vivo clearance of the PEGylated products was investigated. It was found that the generated ant-PEG IgMs induced a clearance of SL as they were intravenously administered with SL. Interestingly, the HIK-M11, generated by PEG-OVA, induced the clearance of both SL and PEG-OVA, while the HIK-M09, generated by SL, induced the clearance of SL only. We here revealed that the presence of serum anti-PEG IgM and the subsequent binding of anti-PEG IgM to the PEGylated products are not necessarily related to the enhanced clearance of the products. It appears that subsequent complement activation following anti-PEG IgM binding is the most important step in dictating the in vivo fate of PEGylated products. This study may have implications for the design, development and clinical application of PEGylated products and therapeutics. - Highlights: • Two monoclonal anti-PEG IgMs

  20. Determination of antibody levels to Candida albicans in healthy and hospitalised adults using a radioimmunoassay

    International Nuclear Information System (INIS)

    Cobb, S.J.; Parratt, D.

    1978-01-01

    A radioimmunoassay for antibody to Candida albicans is described. The test uses whole, killed of organisms as the antigen and radiolabelled sheep anti-human globulins to quantitate different classes of antibody to C. albicans. The assay has been compared with an Ouchterlony precipitin method and found to be simpler, more rapid, and more sensitive than the latter. Results obtained from two groups of symptomless adults indicated that the range of antibody level was wider for a hospitalised group than for a group of blood transfusion donors, particularly in respect of IgG and IgA antibody. The reason for the increase of antibody in hospital patients was not clear but may have been related to antibiotic therapy. The difficulties in interpretation of Candida serology have therefore been re-assessed in the light of more detailed knowledge of the range and type of antibody to be expected in normal individuals. (author)

  1. Immunoglobulin M and G antibody responses to Plasmodium falciparum glutamate-rich protein

    DEFF Research Database (Denmark)

    Dziegiel, M; Rowe, P; Bennett, S

    1993-01-01

    The aims of the present study were to describe the age-related immunoglobulin M (IgM) and IgG response to part of a 220-kDa glutamate-rich protein (GLURP) from Plasmodium falciparum and to determine possible correlations of possession of these antibodies with malaria morbidity. IgM and IgG levels...... in May and in October. Seropositivity rates increased with age to a maximum of 77% for IgM and 95% for IgG in adults. High prevalences of seropositivity were associated with certain human leukocyte antigen class II alleles (DRw8, DR9, DR7, DR4, DQw7, and DQw2) or haplotypes. The relationship between anti......-GLURP489-1271 antibodies and clinical immunity is not clear; asymptomatically infected children aged 5 to 8 years had significantly higher levels of IgG than clinically ill children of the same age, suggesting that antibodies to the carboxy-terminal part of the GLURP may contribute to immunity to P...

  2. Application of solid-phase radioimmunoassay in determining antibodies to Aujeszky's disease virus in blood serum of vaccinated pigs

    International Nuclear Information System (INIS)

    Rodak, L.; Smid, B.; Valicek, L.

    1983-01-01

    In the blood sera of pigs vaccinated with inactivated vaccines manufactured by three different manufacturers the RIA method was used to determine the specific antibodies to the virus of Aujeszky's disease. In certain groups of vaccinated pigs the results of the RIA examination are unfavourably affected by the bond of antibodies to the cellular antigenous determinants. This proves that following vaccination antibodies are formed not only against the viral antigen but also against the antigens of cells on which the vaccination virus is propagated. These shortcomings are eliminated by the use of suitable cellular cultures for the preparation of viral and control antigens. Antigens are applicable for RIA and for ELISA examinations of blood sera of infected and vaccinated pigs. The advantages are described of the RIA and ELISA methods as compared with the virus neutralization test. (author)

  3. Self assertion modeled as a network repertoire of multi-determinant antibodies

    NARCIS (Netherlands)

    Takumi, K.; Boer, R.J. de

    1996-01-01

    We study repertoire selection in a network of natural antibodies that is maintained by stimulatory idiotypic interactions. The natural antibody repertoire develops in an environment of self epitopes to which the self-reactive B cell clones are completely tolerant. For the modeling formalism, we

  4. Determining the feline immunodeficiency virus (FIV) status of FIV-vaccinated cats using point-of-care antibody kits.

    Science.gov (United States)

    Westman, Mark E; Malik, Richard; Hall, Evelyn; Sheehy, Paul A; Norris, Jacqueline M

    2015-10-01

    This study challenges the commonly held view that the feline immunodeficiency virus (FIV) infection status of FIV-vaccinated cats cannot be determined using point-of-care antibody test kits due to indistinguishable antibody production in FIV-vaccinated and naturally FIV-infected cats. The performance of three commercially available point-of-care antibody test kits was compared in a mixed population of FIV-vaccinated (n=119) and FIV-unvaccinated (n=239) cats in Australia. FIV infection status was assigned by considering the results of all antibody kits in concert with results from a commercially available PCR assay (FIV RealPCR™). Two lateral flow immunochromatography test kits (Witness FeLV/FIV; Anigen Rapid FIV/FeLV) had excellent overall sensitivity (100%; 100%) and specificity (98%; 100%) and could discern the true FIV infection status of cats, irrespective of FIV vaccination history. The lateral flow ELISA test kit (SNAP FIV/FeLV Combo) could not determine if antibodies detected were due to previous FIV vaccination, natural FIV infection, or both. The sensitivity and specificity of FIV RealPCR™ for detection of viral and proviral nucleic acid was 92% and 99%, respectively. These results will potentially change the way veterinary practitioners screen for FIV in jurisdictions where FIV vaccination is practiced, especially in shelter scenarios where the feasibility of mass screening is impacted by the cost of testing. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Radioimmunoassay for the quantitative determination of human anti-meningococcal antibodies. Annual report 1 Feb 78-30 Apr 79

    International Nuclear Information System (INIS)

    Gotschlich, E.C.

    1979-01-01

    During the last year of contract support, we have performed a large number of antibody determinations on sera of children vaccinated with Group A or Group C meningococcal polysaccharide. In addition, a number of sera from monkeys vaccinated with different preparations of Group C were also studied. These sera were provided through the efforts of the staff of NIAID and also from other sources

  6. Rapid Proliferation and Differentiation of a Subset of Circulating IgM Memory B Cells to a CpG/Cytokine Stimulus In Vitro.

    Directory of Open Access Journals (Sweden)

    Camilo Vásquez

    Full Text Available Circulating human IgM expressing memory B cells have been incompletely characterized. Here, we compared the phenotype and in vitro functional response (capacity to proliferate and differentiate to antibody secreting cells in response to CpG and a cytokine cocktail (IL-2, IL-6, and IL-10 of sorted naïve B cells, IgM memory B cells and isotype-switched circulating memory B cells. Compared to naïve B cells, IgM memory B cells had lower integrated mean fluorescence intensity (iMFI of BAFF-R, CD38, CD73, and IL-21R, but higher iMFI of CD95, CD11c, TLR9, PD-1, and CD122. Compared to switched memory B cells, IgM memory B cells had higher iMFI of BAFF-R, PD-1, IL-21R, TLR9, and CD122, but lower iMFI of CD38, CD95, and CD73. Four days after receiving the CpG/cytokine cocktail, higher frequencies of IgM than switched memory B cells-and these in turn greater than naïve cells-proliferated and differentiated to antibody secreting cells. At this time point, a small percentage (median of 7.6% of stimulated IgM memory B cells changed isotype to IgG. Thus, among the heterogeneous population of human circulating IgM memory B cells a subset is capable of a rapid functional response to a CpG/cytokine stimulus in vitro.

  7. Tissue Distribution of a Therapeutic Monoclonal Antibody Determined by Large Pore Microdialysis.

    Science.gov (United States)

    Jadhav, Satyawan B; Khaowroongrueng, Vipada; Fueth, Matthias; Otteneder, Michael B; Richter, Wolfgang; Derendorf, Hartmut

    2017-09-01

    Therapeutic monoclonal antibodies (mAbs) exhibit limited distribution to the target tissues. Determination of target tissue interstitial concentration of mAbs is an important aspect in the assessment of their pharmacokinetic/pharmacodynamics relationship especially for mAbs targeting membrane bound receptors. The pharmacokinetics of R7072, a full length mAb (IgG) targeting human insulin-like growth factor-1 receptor was evaluated following a single intravenous dose at 1, 6.25, and 25 mg/kg in healthy female SCID-beige mice. R7072 showed linear pharmacokinetics over the dose range tested and was characterized by low systemic clearance and long terminal half-life. Furthermore, interstitial distribution of R7072 was evaluated in liver, skin, kidney, and muscle tissues using large pore microdialysis (MD) after intravenous administration of 10 mg/kg dose in mice. The relative recoveries of R7072 were consistent and similar between in vitro and in vivo MD experiments. The tissue and interstitial concentrations were significantly lower compared to serum concentrations and found to be highest in liver and lowest in muscle. The interstitial concentrations of R7072 were approximately 2-fold to 4-fold lower than corresponding total tissue concentrations. Large pore MD appears to be an attractive approach for direct measurement of pharmacologically relevant concentrations of therapeutic mAbs in tissue interstitial fluid. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  8. IgG subclass and vaccination stimulus determine changes in antigen specific antibody glycosylation in mice.

    Science.gov (United States)

    Kao, Daniela; Lux, Anja; Schaffert, Anja; Lang, Roland; Altmann, Friedrich; Nimmerjahn, Falk

    2017-12-01

    Immunoglobulin G (IgG) glycosylation can modulate antibody effector functions. Depending on the precise composition of the sugar moiety attached to individual IgG glycovariants either pro- or anti-inflammatory effector pathways can be initiated via differential binding to type I or type II Fc-receptors. However, an in depth understanding of how individual IgG subclasses are glycosylated during the steady state and how their glycosylation pattern changes during vaccination is missing. To monitor IgG subclass glycosylation during the steady state and upon vaccination of mice with different T-cell dependent and independent antigens, tryptic digests of serum, and antigen-specific IgG preparations were analyzed by reversed phase-liquid chromatography-mass spectrometry. We show that there is a remarkable difference with respect to how individual IgG subclasses are glycosylated during the steady state. More importantly, upon T-cell dependent and independent vaccinations, individual antigen-specific IgG subclasses reacted differently with respect to changes in individual glycoforms, suggesting that the IgG subclass itself is a major determinant of restricting or allowing alterations in specific IgG glycovariants. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Hyper IgM Syndrome with low IgM and thrombocytosis: an unusual case of immunodeficiency.

    Science.gov (United States)

    Yousef, Ejaz; Arshad Alvi, M

    2016-09-01

    We report a 5 years old male child with low serum IgG, IgA and IgM levels, who presented with recurrent perianal and oral ulcers, intermittent fever, and protracted diarrhea. Despite the lack of typical respiratory symptoms, low serum IgM level and persistent thrombocytosis, an X-linked hyper-IgM syndrome (X-HIGM) was considered. Laboratory investigations revealed a diagnosis of hyper-IgM syndrome caused by CD40L deficiency.

  10. Determination of neuronal antibodies in suspected and definite Creutzfeldt-Jakob disease.

    Science.gov (United States)

    Grau-Rivera, Oriol; Sánchez-Valle, Raquel; Saiz, Albert; Molinuevo, José Luis; Bernabé, Reyes; Munteis, Elvira; Pujadas, Francesc; Salvador, Antoni; Saura, Júlia; Ugarte, Antonio; Titulaer, Maarten; Dalmau, Josep; Graus, Francesc

    2014-01-01

    Creutzfeldt-Jakob disease (CJD) and autoimmune encephalitis with antibodies against neuronal surface antigens (NSA-abs) may present with similar clinical features. Establishing the correct diagnosis has practical implications in the management of care for these patients. To determine the frequency of NSA-abs in the cerebrospinal fluid of patients with suspected CJD and in patients with pathologically confirmed (ie, definite) CJD. A mixed prospective (suspected) and retrospective (definite) CJD cohort study was conducted in a reference center for detection of NSA-abs. The population included 346 patients with suspected CJD and 49 patients with definite CJD. Analysis of NSA-abs in cerebrospinal fluid with brain immunohistochemistry optimized for cell-surface antigens was performed. Positive cases in the suspected CJD group were further studied for antigen specificity using cell-based assays. All definite CJD cases were comprehensively tested for NSA-abs, with cell-based assays used for leucine-rich glioma-inactivated 1 (LGI1), contactin-associated protein-like 2 (CASPR2), N-methyl-d-aspartate (NMDA), and glycine (GlY) receptors. Neuronal surface antigens were detected in 6 of 346 patients (1.7%) with rapid neurologic deterioration suggestive of CJD. None of these 6 patients fulfilled the diagnostic criteria for probable or possible CJD. The target antigens included CASPR2, LGI1, NMDAR, aquaporin 4, Tr (DNER [δ/notch-like epidermal growth factor-related receptor]), and an unknown protein. Four of the patients developed rapidly progressive dementia, and the other 2 patients had cerebellar ataxia or seizures that were initially considered to be myoclonus without cognitive decline. The patient with Tr-abs had a positive 14-3-3 test result. Small cell lung carcinoma was diagnosed in the patient with antibodies against an unknown antigen. All patients improved or stabilized after appropriate treatment. None of the 49 patients with definite CJD had NSA-abs. A low, but

  11. The clinical significance of antibody determination to cyclic citrullinated peptides in systemic sclerosis

    Directory of Open Access Journals (Sweden)

    Stamenković Bojana

    2012-01-01

    Full Text Available Introduction. Anti-citrullinated peptides antibodies (ACPA are present in 80% of sera of rheumatoid arthritis (RA patients with high specificity for diagnosis and prediction for the development of early erosive arthritis. A few studies have reported a low frequency ACPA in systemic sclerosis (SSc patients with the presence of arthritis. Objective. The aim of our study was to determine the frequency of ACPA in systemic sclerosis (SSc patients, their correlation with clinical manifestations and radiographic features. Methods. The study included 82 patients with SSc, mean age 54.4 years, 59 with the limited (lSSc and 23 with the diffuse (dSSc form of the disease. The control group included 28 healthy age and sex matched subjects. ACPA and rheumatoid factor (RF were determined in all SSc patients and healthy subjects in whom standard radiography of hands and wrists was also done. Results. The presence of ACPA was detected in 11 (13.4% of SSc patients. Their level was not increased in any of the controls. Positive RF was found in 15.9% of SSc patients. Arthritis was present in 17.1%, as well as marginal bone erosions. There was a statistically significant association between positive ACPA and arthritis (p<0.0001 and positive ACPA and marginal bone erosions (p=0.0002. Conclusion. The research confirmed the correlation between ACPA with clinical signs of arthritis and radiographic damage of hand joints. ACPA is a useful diagnostic marker in the identification of SSc patients with arthritis and anatomic bone damage enabling the use of adequate therapy in order to prevent joint damage and poor quality of life.

  12. Prevalence of antibody to hantaviruses in humans and rodents in the Caribbean region of Colombia determined using Araraquara and Maciel virus antigens

    Directory of Open Access Journals (Sweden)

    Camilo Guzmán

    2013-04-01

    Full Text Available We tested sera from 286 agricultural workers and 322 rodents in the department of Córdoba, northeastern Colombia, for antibodies against two hantaviruses. The sera were analysed by indirect ELISA using the lysate of Vero E6 cells infected with Maciel virus (MACV or the N protein of Araraquara virus (ARAV as antigens for the detection of antibodies against hantaviruses. Twenty-four human sera were IgG positive using one or both antigens. We detected anti-MACV IgG antibodies in 10 sera (3.5% and anti-ARAV antibodies in 21 sera (7.34%. Of the 10 samples that were positive for MACV, seven (70% were cross-reactive with ARAV; seven of the 21 ARAV-positive samples were cross-reactive with MACV. Using an ARAV IgM ELISA, two of the 24 human sera (8.4% were positive. We captured 322 rodents, including 210 Cricetidae (181 Zygodontomys brevicauda, 28 Oligoryzomys fulvescens and 1 Oecomys trinitatis, six Heteromys anomalus (Heteromyidae, one Proechimys sp. (Echimyidae and 105 Muridae (34 Rattus rattus and 71 Mus musculus. All rodent sera were negative for both antigens. The 8.4% detection rate of hantavirus antibodies in humans is much higher than previously found in serosurveys in North America, suggesting that rural agricultural workers in northeastern Colombia are frequently exposed to hantaviruses. Our results also indicate that tests conducted with South American hantavirus antigens could have predictive value and could represent a useful alternative for the diagnosis of hantavirus infection in Colombia.

  13. An overview of Western blotting for determining antibody specificities for immunohistochemistry.

    Science.gov (United States)

    Kurien, Biji T; Dorri, Yaser; Dillon, Skyler; Dsouza, Anil; Scofield, R Hal

    2011-01-01

    Despite its overall simplicity, protein blotting or Western blotting has been proven to be a powerful procedure for the immunodetection of proteins, especially those that are of low abundance, following electrophoresis. The usefulness of this procedure stems from its ability to provide simultaneous resolution of multiple immunogenic antigens within a sample for detection by specific antibodies. Protein blotting has evolved greatly since its inception and researchers have a variety of ways and means to carry out this transfer. This procedure is used in combination with other important antibody-based detection methods such as enzyme-linked immunosorbant assay and immunohistochemistry to provide confirmation of results both in research and diagnostic testing. Specificity of antibodies used for immunohistochemistry is of critical importance and therefore Western blot is a "must" to address antibodies' specificity.

  14. Cross-reactive Carbohydrate Determinant Contributes to the False Positive IgE Antibody to Peanut

    Directory of Open Access Journals (Sweden)

    Komei Ito

    2005-01-01

    Conclusions: Social education about the features of peanut allergy is needed in Japan. Anti-CCD IgE antibody was suggested to be one of the mechanisms contributing to the false positive detection of peanut IgE. Detection of anti-HRP or anti-bromelain IgE can be a useful tool to recognize the presence of anti-CCD antibodies.

  15. Pneumococcal polysaccharide vaccination elicits IgG anti-AB blood group antibodies in healthy individuals and patients with Type I diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Wendelin Wolfram

    2016-11-01

    Full Text Available Hypothesis: Blood group antibodies are natural antibodies that develop early in life in response to cross-reactive environmental antigens in the absence of antigen encounter. Even later in life structural similarities in saccharide composition between environmental antigens such as bacterial polysaccharides and blood group A/B antigens could lead to changes in serum levels, IgM/IgG isotype and affinity maturation of blood group anti-A/B antibodies. We adressed the question whether immunization with pneumococcal polysaccharide (PnP vaccine (PPV Pneumovax®23 could have such an effect in patients with with type I diabetes mellitus (DM I, an autoimmune disease where an aberrant immune response to microbial antigens likely plays a role.Methods: Anti-PnP IgM and IgG responses were determined by ELISA and the Diamed-ID Micro Typing System was used to screen anti-A/B antibody titer before and after Pneumovax®23 immunization in 28 healthy individuals and 16 patients with DM I. In addition, surface plasmon resonance (SPR technology using the Biacore® device and a synthetic blood group A/B trisaccharide as the antigen was applied to investigate IgM and IgG anti-A/B antibodies and to measure antibody binding dynamics. Results: All healthy individuals and DM I patients responded with anti-PnP IgM and IgG antibody production four to six weeks after Pneumovax®23 (Pn23 immunization, while no increase in blood group anti-A/B antibody titer was observed when measured by the Diamed-ID Micro Typing System. Interestingly, isotype-specific testing by SPR-technology revealed an increase in blood group anti-A/B IgG, but not IgM, following Pn23 immunization in both patients and controls. No change in binding characteristics of blood group anti-A/B antibodies could be detected following Pn23 vaccination, supporting the assumption of an increase in IgG antibody titer with no or very little affinity maturation.Conclusion: The study provides evidence for epitope sharing

  16. Timing of the human prenatal antibody response to Plasmodium falciparum antigens.

    Directory of Open Access Journals (Sweden)

    Samuel Tassi Yunga

    Full Text Available Plasmodium falciparum (Pf-specific T- and B-cell responses may be present at birth; however, when during fetal development antibodies are produced is unknown. Accordingly, cord blood samples from 232 preterm (20-37 weeks of gestation and 450 term (≥37 weeks babies were screened for IgM to Pf blood-stage antigens MSP1, MSP2, AMA1, EBA175 and RESA. Overall, 25% [95% CI = 22-28%] of the 682 newborns were positive for IgM to ≥1 Pf antigens with the earliest response occurring at 22 weeks. Interestingly, the odds of being positive for cord blood Pf IgM decreased with gestational age (adjusted OR [95% CI] at 20-31 weeks = 2.55 [1.14-5.85] and at 32-36 weeks = 1.97 [0.92-4.29], with ≥37 weeks as reference; however, preterm and term newborns had similar levels of Pf IgM and recognized a comparable breadth of antigens. Having cord blood Pf IgM was associated with placental malaria (adjusted OR [95% CI] = 2.37 [1.25-4.54]. To determine if in utero exposure occurred via transplacental transfer of Pf-IgG immune complexes (IC, IC containing MSP1 and MSP2 were measured in plasma of 242 mother-newborn pairs. Among newborns of IC-positive mothers (77/242, the proportion of cord samples with Pf IC increased with gestational age but was not associated with Pf IgM, suggesting that fetal B cells early in gestation had not been primed by IC. Finally, when cord mononuclear cells from 64 term newborns were cultured in vitro, only 11% (7/64 of supernatants had Pf IgM; whereas, 95% (61/64 contained secreted Pf IgG. These data suggest fetal B cells are capable of making Pf-specific IgM from early in the second trimester and undergo isotype switching to IgG towards term.

  17. N+1 Engineering of an Aspartate Isomerization Hotspot in the Complementarity-Determining Region of a Monoclonal Antibody.

    Science.gov (United States)

    Patel, Chetan N; Bauer, Scott P; Davies, Julian; Durbin, Jim D; Shiyanova, Tatiyana L; Zhang, Kai; Tang, Jason X

    2016-02-01

    Aspartate (Asp) isomerization is a common degradation pathway and a potential critical quality attribute that needs to be well characterized during the optimization and development of therapeutic antibodies. A putative Asp-serine (Ser) isomerization motif was identified in the complementarity-determining region of a humanized monoclonal antibody and shown to be a developability risk using accelerated stability analyses. To address this issue, we explored different antibody engineering strategies. Direct engineering of the Asp residue resulted in a greater than 5× loss of antigen-binding affinity and bioactivity, indicating a critical role for this residue. In contrast, rational engineering of the Ser residue at the n+1 position had a negligible impact on antigen binding affinity and bioactivity compared with the parent molecule. Furthermore, the n+1 engineering strategy effectively eliminated Asp isomerization as determined by accelerated stability analysis. This outcome affirms that the rate of Asp isomerization is strongly dependent on the identity of the n+1 residue. This report highlights a systematic antibody engineering strategy for mitigating an Asp isomerization developability risk during lead optimization. Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  18. Determining the binding affinity of therapeutic monoclonal antibodies towards their native unpurified antigens in human serum.

    Science.gov (United States)

    Bee, Christine; Abdiche, Yasmina N; Pons, Jaume; Rajpal, Arvind

    2013-01-01

    Monoclonal antibodies (mAbs) are a growing segment of therapeutics, yet their in vitro characterization remains challenging. While it is essential that a therapeutic mAb recognizes the native, physiologically occurring epitope, the generation and selection of mAbs often rely on the use of purified recombinant versions of the antigen that may display non-native epitopes. Here, we present a method to measure both, the binding affinity of a therapeutic mAb towards its native unpurified antigen in human serum, and the antigen's endogenous concentration, by combining the kinetic exclusion assay and Biacore's calibration free concentration analysis. To illustrate the broad utility of our method, we studied a panel of mAbs raised against three disparate soluble antigens that are abundant in the serum of healthy donors: proprotein convertase subtilisin/kexin type 9 (PCSK9), progranulin (PGRN), and fatty acid binding protein (FABP4). We also determined the affinity of each mAb towards its purified recombinant antigen and assessed whether the interactions were pH-dependent. Of the six mAbs studied, three did not appear to discriminate between the serum and recombinant forms of the antigen; one mAb bound serum antigen with a higher affinity than recombinant antigen; and two mAbs displayed a different affinity for serum antigen that could be explained by a pH-dependent interaction. Our results highlight the importance of taking pH into account when measuring the affinities of mAbs towards their serum antigens, since the pH of serum samples becomes increasingly alkaline upon aerobic handling.

  19. An experimental design approach to optimize an amperometric immunoassay on a screen printed electrode for Clostridium tetani antibody determination.

    Science.gov (United States)

    Patris, Stéphanie; Vandeput, Marie; Kenfack, Gersonie Momo; Mertens, Dominique; Dejaegher, Bieke; Kauffmann, Jean-Michel

    2016-03-15

    An immunoassay for the determination of anti-tetani antibodies has been developed using a screen printed electrode (SPE) as solid support for toxoid (antigen) immobilization. The assay was performed in guinea pig serum. The immunoreaction and the subsequent amperometric detection occurred directly onto the SPE surface. The assay consisted of spiking the anti-tetani sample directly onto the toxoid modified SPE, and then a second antibody, i.e. a HRP-labeled anti-immunoglobulin G, was deposited onto the biosensor. Subsequent amperometric detection was realized by spiking 10 µL of a hydroquinone (HQ) solution into 40 µL of buffer solution containing hydrogen peroxide. An experimental design approach was implemented for the optimization of the immunoassay. The variables of interest, such as bovine serum albumin (BSA) concentration, incubation times and labeled antibody dilution, were optimized with the aid of the response surface methodology using a circumscribed central composite design (CCCD). It was observed that two factors exhibited the greatest impact on the response, i.e. the anti-tetani incubation time and the dilution factor of the labeled antibody. It was discovered that in order to maximize the response, the dilution factor should be small, while the anti-tetani antibody incubation time should be long. The BSA concentration and the HRP-anti-IgG incubation had very limited influence. Under the optimized conditions, the immunoassay had a limit of detection of 0.011 IU/mL and a limit of quantification of 0.012 IU/mL. These values were below the protective human antibody limit of 0.06 IU/mL. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. [Human anti-murine immunoglobulin antibodies as disturbing factors in TSH determination].

    Science.gov (United States)

    Horneff, G; Becker, W; Wolf, F; Kalden, J R; Burmester, G R

    1991-03-18

    Monoclonal murine antibodies are increasingly used for immunotherapy and in vivo diagnostic procedures such as immunoscintigraphy. The therapeutic or diagnostic reagent however, is a foreign antigen, which may induce host reactivity. This may interfere with the therapeutic or diagnostic reagent in vivo, resulting in a loss of efficacy or the necessity to increase dosages. In addition, there is an important interference to in vitro immunoassays detecting specific antigens utilizing murine monoclonal antibodies. In the present study, sera of patients who had undergone a therapeutic trial using 140 mg of an anti-CD4 antibody, were investigated. Human anti-murine-immunoglobulin-antibodies (HAMA) were detected 2-3 weeks after treatment was started and reached maximal amounts of 0.8 micrograms/ml after a single and 2 micrograms/ml after a repeated treatment course. Parallely raised values of TSH were found in sera containing HAMAs of more than 0.3 micrograms/ml. Elevations of TSH levels up to 13 microU/ml were most pronounced after a repeated trial of the murine antibody and were detectable up to 20 weeks.

  1. Determining infants' age for measles vaccination based on persistence of protective level of maternal measles antibody.

    Science.gov (United States)

    Shilpi, Tanjida; Sattar, Humayun; Miah, Md Ruhul Amin

    2009-12-01

    The present study was conducted over a period of one year to find the right time for measles vaccination when maternal antibody titer in infants was decayed rendering them susceptible to wild virus infection. Blood samples were collected from the cord of new born (147), 2-5 months (47) and 5 to 7.5 months (24) of age. The mean measles IgG antibody titer detected in cord blood at birth (0 months) was 348.8 mlU/mL which steeply decreased to 155.6 mlU/mL by the age of 2-3 months. After that the fall in antibody becomes relatively slower and decreased to 101.6 mIU/mL by the age of 3-5 months and 38.8 mlU/mL by the age of 5-6 months and to 19.2 mIU/mL between the age of 6 to 7.5 months. The fall in antibody level with the advance of age was statistically significant (p < 0.001 ). Majority of the subjects (97.6%) exhibited protective level of antibody at birth. But only a little above one-quarter (25.5%) of them persisted the protective level between the age of 2-5 months and none had protective level from 5 months onwards.

  2. Lack of immunoglobulin M suppression by immunoglobulin G antibody in thymectomized, irradiated, and bone marrow-reconstituted mice infected with Toxoplasma gondii.

    OpenAIRE

    Aryanpour, J; Hafizi, A; Modabber, F

    1980-01-01

    Thymectomized, irradiated, bone marrow-reconstituted (T-deprived) mie infected with an avirulent strain of Toxoplasma gondii produced antibody titers comparable to those produced in intact syngeneic mice. Both immunoglobulin M (IgM) and IgG antibodies were produced in T-deprived animals; however, the IgM antibody remained constant in the presence of increasing amounts of IgG. In the intact animals, IgM became undetectable by day 50 postinfection as expected. Feedback inhibition of IgM by IgG ...

  3. Lack of immunoglobulin M suppression by immunoglobulin G antibody in thymectomized, irradiated, and bone marrow-reconstituted mice infected with Toxoplasma gondii.

    Science.gov (United States)

    Aryanpour, J; Hafizi, A; Modabber, F

    1980-03-01

    Thymectomized, irradiated, bone marrow-reconstituted (T-deprived) mie infected with an avirulent strain of Toxoplasma gondii produced antibody titers comparable to those produced in intact syngeneic mice. Both immunoglobulin M (IgM) and IgG antibodies were produced in T-deprived animals; however, the IgM antibody remained constant in the presence of increasing amounts of IgG. In the intact animals, IgM became undetectable by day 50 postinfection as expected. Feedback inhibition of IgM by IgG seems to be dependent upon T-cells in Toxoplasma-infected mice.

  4. Antiprothrombin Antibodies

    Directory of Open Access Journals (Sweden)

    Polona Žigon

    2015-05-01

    Full Text Available In patients with the antiphospholipid syndrome (APS, the presence of a group of pathogenic autoantibodies called antiphospholipid antibodies causes thrombosis and pregnancy complications. The most frequent antigenic target of antiphospholipid antibodies are phospholipid bound β2-glycoprotein 1 (β2GPI and prothrombin. The international classification criteria for APS connect the occurrence of thrombosis and/or obstetric complications together with the persistence of lupus anticoagulant, anti-cardiolipin antibodies (aCL and antibodies against β2GPI (anti-β2GPI into APS. Current trends for the diagnostic evaluation of APS patients propose determination of multiple antiphospholipid antibodies, among them also anti-prothrombin antibodies, to gain a common score which estimates the risk for thrombosis in APS patients. Antiprothrombin antibodies are common in APS patients and are sometimes the only antiphospholipid antibodies being elevated. Methods for their determination differ and have not yet been standardized. Many novel studies confirmed method using phosphatidylserine/prothrombin (aPS/PT ELISA as an antigen on solid phase encompass higher diagnostic accuracy compared to method using prothrombin alone (aPT ELISA. Our research group developed an in-house aPS/PT ELISA with increased analytical sensitivity which enables the determination of all clinically relevant antiprothrombin antibodies. aPS/PT exhibited the highest percentage of lupus anticoagulant activity compared to aCL and anti-β2GPI. aPS/PT antibodies measured with the in-house method associated with venous thrombosis and presented the strongest independent risk factor for the presence of obstetric complications among all tested antiphospholipid antibodies

  5. Oral antibiotics enhance antibody responses to keyhole limpet hemocyanin in orally but not muscularly immunized chickens.

    Science.gov (United States)

    Murai, Atsushi; Kitahara, Kazuki; Okumura, Shouta; Kobayashi, Misato; Horio, Fumihiko

    2016-02-01

    Recent studies have emphasized the crucial role of gut microbiota in triggering and modulating immune response. We aimed to determine whether the modification of gut microbiota by oral co-administration of two antibiotics, ampicillin and neomycin, would lead to changes in the antibody response to antigens in chickens. Neonatal chickens were given or not given ampicillin and neomycin (0.25 and 0.5 g/L, respectively) in drinking water. At 2 weeks of age, the chicks were muscularly or orally immunized with antigenic keyhole limpet hemocyanin (KLH), and then serum anti-KLH antibody levels were examined by ELISA. In orally immunized chicks, oral antibiotics treatment enhanced antibody responses (IgM, IgA, IgY) by 2-3-fold compared with the antibiotics-free control, while the antibiotics did not enhance antibody responses in the muscularly immunized chicks. Concomitant with their enhancement of antibody responses, the oral antibiotics also lowered the Lactobacillus species in feces. Low doses of antibiotics (10-fold and 100-fold lower than the initial trial), which failed to change the fecal Lactobacillus population, did not modify any antibody responses when chicks were orally immunized with KLH. In conclusion, oral antibiotics treatment enhanced the antibody response to orally exposed antigens in chickens. This enhancement of antibody response was associated with a modification of the fecal Lactobacillus content, suggesting a possible link between gut microbiota and antibody response in chickens. © 2015 Japanese Society of Animal Science.

  6. Genetic variants are major determinants of CSF antibody levels in multiple sclerosis

    DEFF Research Database (Denmark)

    Goris, An; Pauwels, Ine; Gustavsen, Marte W

    2015-01-01

    Immunological hallmarks of multiple sclerosis include the production of antibodies in the central nervous system, expressed as presence of oligoclonal bands and/or an increased immunoglobulin G index-the level of immunoglobulin G in the cerebrospinal fluid compared to serum. However, the underlying...... differences between oligoclonal band-positive and -negative patients with multiple sclerosis and reasons for variability in immunoglobulin G index are not known. To identify genetic factors influencing the variation in the antibody levels in the cerebrospinal fluid in multiple sclerosis, we have performed...... of being oligoclonal band positive and 7.75% of the variation in immunoglobulin G index. Both traits are associated with clinical features of disease such as female gender, age at onset and severity. This is the largest study population so far investigated for the genetic influence on antibody levels...

  7. Bead injection ELISA for the determination of antibodies implicated in type 1 diabetes mellitus.

    Science.gov (United States)

    Carroll, Andrea D; Scampavia, Louis; Luo, Dong; Lernmark, Ake; Ruzicka, Jaromir

    2003-09-01

    This work introduces a novel analytical method for the detection and study of GAD65 autoantibodies, which have been implicated in the onset of type 1 diabetes. There is a clinical need for a rapid and automated assay for GAD65 autoantibodies. Therefore, this method was designed to exploit the advantages of bead injection (BI) analysis for enzyme-linked immunosorbent assays (ELISA). BI ELISA is a microscale technique that uses enzyme labeled secondary antibodies to detect the capture of target antibodies on immobilized antigen in the flow cell of the lab-on-valve (LOV) manifold. A detection limit of 20 ng mL(-1) of GAD65 monoclonal antibody 144 compares favorably with the sensitivity and precision of a standard ELISA currently employed to detect GAD65 autoantibodies. Compared to the standard ELISA protocol, BI ELISA offers a significantly reduced assay time and complete automation of solution handling and detection.

  8. Evaluation of a rapid IgM detection test for diagnosis of acute leptospirosis in dogs.

    Science.gov (United States)

    Lizer, J; Grahlmann, M; Hapke, H; Velineni, S; Lin, D; Kohn, B

    2017-05-27

    Recently, a lateral flow assay (LFA) for detection of Leptospira -specific IgM in canine sera became commercially available in Europe. The present study aims to evaluate the diagnostic performance of this assay using canine sera from a collection of diagnostic accessions. Diagnostic sensitivity was assessed by testing 37 acute-phase and 9 corresponding convalescent-phase sera from dogs with a confirmed diagnosis of leptospirosis. Specificity was determined by testing sera from sick dogs with non-leptospiral infections (n=15) and healthy dogs with incomplete history of vaccination (n=45). During acute phase of illness, LFA scored positive for 28/37 sera with a sensitivity of 75.7 per cent while only 9/37 (24.3 per cent) samples were positive on microscopic agglutination test. The specificity of the LFA was 98.3 per cent (59/60). This test showed 89.7 and 100 per cent overall agreements with clinical diagnosis for acute-phase and convalescent-phase sera, respectively. The impact of vaccination on the LFA was also determined and vaccine-stimulated IgM responses were negative in 19/25 (76 per cent) dogs at 12 weeks post vaccination. In conclusion, the LFA is a rapid and reliable test for early detection of Leptospira -specific IgM during acute phase of canine leptospirosis. However, interpretation of a positive result must be made in the context of clinical signs and vaccination history. British Veterinary Association.

  9. Antibody complementarity-determining regions (CDRs can display differential antimicrobial, antiviral and antitumor activities.

    Directory of Open Access Journals (Sweden)

    Luciano Polonelli

    Full Text Available BACKGROUND: Complementarity-determining regions (CDRs are immunoglobulin (Ig hypervariable domains that determine specific antibody (Ab binding. We have shown that synthetic CDR-related peptides and many decapeptides spanning the variable region of a recombinant yeast killer toxin-like antiidiotypic Ab are candidacidal in vitro. An alanine-substituted decapeptide from the variable region of this Ab displayed increased cytotoxicity in vitro and/or therapeutic effects in vivo against various bacteria, fungi, protozoa and viruses. The possibility that isolated CDRs, represented by short synthetic peptides, may display antimicrobial, antiviral and antitumor activities irrespective of Ab specificity for a given antigen is addressed here. METHODOLOGY/PRINCIPAL FINDINGS: CDR-based synthetic peptides of murine and human monoclonal Abs directed to: a a protein epitope of Candida albicans cell wall stress mannoprotein; b a synthetic peptide containing well-characterized B-cell and T-cell epitopes; c a carbohydrate blood group A substance, showed differential inhibitory activities in vitro, ex vivo and/or in vivo against C. albicans, HIV-1 and B16F10-Nex2 melanoma cells, conceivably involving different mechanisms of action. Antitumor activities involved peptide-induced caspase-dependent apoptosis. Engineered peptides, obtained by alanine substitution of Ig CDR sequences, and used as surrogates of natural point mutations, showed further differential increased/unaltered/decreased antimicrobial, antiviral and/or antitumor activities. The inhibitory effects observed were largely independent of the specificity of the native Ab and involved chiefly germline encoded CDR1 and CDR2 of light and heavy chains. CONCLUSIONS/SIGNIFICANCE: The high frequency of bioactive peptides based on CDRs suggests that Ig molecules are sources of an unlimited number of sequences potentially active against infectious agents and tumor cells. The easy production and low cost of small

  10. A comparison of Rubazyme-M and MACRIA for the detection of rubella-specific IgM

    International Nuclear Information System (INIS)

    Best, J.M.; Palmer, S.J.; Morgan-Capner, P.; Hodgson, J.

    1984-01-01

    One hundred and eighty-six carefully selected sera were tested for rubella-specific IgM by Rubazyme-M (Abbott Diagnostics) and an M-antibody capture radioimmunoassay (MACRIA). Eleven of these sera were from cases of infectious mononucleosis, six of which gave positive results in MACRIA, while one gave a positive result in Rubazyme-M. Of the remaining 175 sera, 158 gave concordant results whilst 17 sera gave discordant results; these 17 were also tested by serum fractionation. Problems were encountered with all assay systems used. It is therefore recommended that the results of all tests for rubella-specific IgM should be interpreted with caution. (Auth.)

  11. Indirect immunofluorescence assay for the simultaneous detection of antibodies against clinically important old and new world hantaviruses.

    Directory of Open Access Journals (Sweden)

    Sabine Lederer

    Full Text Available In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV, Puumala (PUUV, Seoul (SEOV, Saaremaa (SAAV, Dobrava (DOBV, Sin Nombre (SNV or Andes (ANDV. For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n=97; SEOV (China, n=5; DOBV (Romania, n=7; SNV (Canada, n=23; ANDV (Argentina and Chile, n=52. The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n=177; 96% and/or IgM (n=131; 72%, while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99% of the patient sera were IgG and 131 (71% IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%. Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%. Of the 89 control sera, 2 (2% showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in >90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV.

  12. Selection of apoptotic cell specific human antibodies from adult bone marrow.

    Directory of Open Access Journals (Sweden)

    Caroline Grönwall

    Full Text Available Autoreactive antibodies that recognize neo-determinants on apoptotic cells in mice have been proposed to have protective, homeostatic and immunoregulatory properties, although our knowledge about the equivalent antibodies in humans has been much more limited. In the current study, human monoclonal antibodies with binding specificity for apoptotic cells were isolated from the bone marrow of healthy adults using phage display technology. These antibodies were shown to recognize phosphorylcholine (PC-associated neo-determinants. Interestingly, three of the four identified apoptotic cell-specific antibody clones were encoded by VH3 region rearrangements with germline or nearly germline configuration without evidence of somatic hypermutation. Importantly, the different identified antibody clones had diverse heavy chain CDR3 and deduced binding surfaces as suggested by structure modeling. This may suggest a potentially great heterogeneity in human antibodies recognizing PC-related epitopes on apoptotic cells. To re-construct the postulated structural format of the parental anti-PC antibody, the dominant clone was also expressed as a recombinant human polymeric IgM, which revealed a substantially increased binding reactivity, with dose-dependent and antigen-inhibitable binding of apoptotic cells. Our findings may have implication for improved prognostic testing and therapeutic interventions in human inflammatory disease.

  13. Improved Diagnosis of Acute Pulmonary Histoplasmosis by Combining Antigen and Antibody Detection.

    Science.gov (United States)

    Richer, Sarah M; Smedema, Melinda L; Durkin, Michelle M; Herman, Katie M; Hage, Chadi A; Fuller, Deanna; Wheat, L Joseph

    2016-04-01

    Acute pulmonary histoplasmosis can be severe, especially following heavy inoculum exposure. Rapid diagnosis is critical and often possible by detection of antigen, but this test may be falsely negative in 17% of such cases. Antibody detection by enzyme immunoassay (EIA) may increase sensitivity and permit the measurement of immunoglobulin M (IgM) and immunoglobulin G (IgG) classes of antibodies separately. Microplates coated with Histoplasma antigen were used for testing of serum from patients with acute pulmonary histoplasmosis and controls in the MVista Histoplasma antibody EIA. Results for IgG and IgM were reported independently. IgG antibodies were detected in 87.5%, IgM antibodies in 67.5%, and IgG and/or IgM antibodies in 88.8% of patients with acute pulmonary histoplasmosis in this assay, while immunodiffusion, complement fixation, and antigen testing showed sensitivities of 55.0%, 73.1%, and 67.5%, respectively (n = 80). Combining antigen and antibody detection increased the sensitivity to 96.3%. The MVista Histoplasma antibody EIA offers increased sensitivity over current antibody tests while also allowing separate detection of IgG and IgM antibodies and complementing antigen detection. Combining antigen and EIA antibody testing provides an optimal method for diagnosis of acute pulmonary histoplasmosis. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America.

  14. Origin, diversity and maturation of human antiviral antibodies analyzed by high-throughput sequencing

    Directory of Open Access Journals (Sweden)

    Ponraj ePrabakaran

    2012-08-01

    Full Text Available Our understanding of how antibodies are generated and function could help develop effective vaccines and antibody-based therapeutics against viruses such as HIV-1, SARS Coronavirus (CoV, and Hendra and Nipah viruses (henipaviruses. Although broadly neutralizing antibodies (bnAbs against the HIV-1 were observed in patients, elicitation of such bnAbs remains a major challenge when compared to other viral targets. We previously hypothesized that HIV-1 could have evolved a strategy to evade the immune system due to absent or very weak binding of germline antibodies to the conserved epitopes that may not be sufficient to initiate and/or maintain an effective immune response. To further explore our hypothesis, we used the 454 sequence analysis of a large naïve library of human IgM antibodies which had been used for selecting antibodies against SARS Coronavirus (CoV receptor-binding domain (RBD, and soluble G proteins (sG of Hendra and Nipah viruses (henipaviruses. We found that the human IgM repertoires from the 454 sequencing have diverse germline usages, recombination patterns, junction diversity and a lower extent of somatic mutation. In this study, we identified germline intermediates of antibodies specific to HIV-1 and other viruses as observed in normal individuals, and compared their genetic diversity and somatic mutation level along with available structural and functional data. Further computational analysis will provide framework for understanding the underlying genetic and molecular determinants related to maturation pathways of antiviral bnAbs that could be useful for applying novel approaches to the design of effective vaccine immunogens and antibody-based therapeutics.

  15. Stochastic simulation modeling to determine time to detect Bovine Viral Diarrhea antibodies in bulk tank milk

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Enøe, Claes; Krogh, Kaspar

    2014-01-01

    A stochastic simulation model was developed to estimate the time from introduction ofBovine Viral Diarrhea Virus (BVDV) in a herd to detection of antibodies in bulk tank milk(BTM) samples using three ELISAs. We assumed that antibodies could be detected, after afixed threshold prevalence of seroco......A stochastic simulation model was developed to estimate the time from introduction ofBovine Viral Diarrhea Virus (BVDV) in a herd to detection of antibodies in bulk tank milk(BTM) samples using three ELISAs. We assumed that antibodies could be detected, after afixed threshold prevalence...... of seroconverted milking cows was reached in the herd. Differentthresholds were set for each ELISA, according to previous studies. For each test, antibodydetection was simulated in small (70 cows), medium (150 cows) and large (320 cows)herds. The assays included were: (1) the Danish blocking ELISA, (2......, which was the most efficient ELISA, could detect antibodiesin the BTM of a large herd 280 days (95% prediction interval: 218; 568) after a transientlyinfected (TI) milking cow has been introduced into the herd. The estimated time to detectionafter introduction of one PI calf was 111 days (44; 605...

  16. Determining Vaccination Frequency in Farmed Rainbow Trout Using Vibrio anguillarum O1 Specific Serum Antibody Measurements

    DEFF Research Database (Denmark)

    Holten-Andersen, Lars; Dalsgaard, Inger; Nylén, Jørgen

    2012-01-01

    Background Despite vaccination with a commercial vaccine with a documented protective effect against Vibrio anguillarum O1 disease outbreaks caused by this bacterium have been registered among rainbow trout at Danish fish farms. The present study examined specific serum antibody levels as a valid...

  17. Determination of neuronal antibodies in suspected and definite Creutzfeldt-Jakob disease

    NARCIS (Netherlands)

    O. Grau-Rivera (Oriol); R. Sánchez-Valle (Raquel); A. Saiz (Albert Abe); J.L. Molinuevo (José Luis); R. Bernabé (Reyes); E. Munteis (Elvira); F. Pujadas (Francesc); A. Salvador (Antoni); J. Saura (Júlia); A. Ugarte (Antonio); M.J. Titulaer (Maarten); J. Dalmau (Josep); F. Graus (Francesc)

    2014-01-01

    textabstractIMPORTANCE: Creutzfeldt-Jakob disease (CJD) and autoimmune encephalitis with antibodies against neuronal surface antigens (NSA-abs) may present with similar clinical features. Establishing the correct diagnosis has practical implications in the management of care for these patients.

  18. Human anti-mouse antibody response induced by anti-CD4 monoclonal antibody therapy in patients with rheumatoid arthritis.

    Science.gov (United States)

    Horneff, G; Winkler, T; Kalden, J R; Emmrich, F; Burmester, G R

    1991-04-01

    The development of human anti-mouse monoclonal antibodies (HAMAs) was investigated in 10 patients with rheumatoid arthritis (RA) who had undergone an experimental therapeutic trial with an anti-CD4 monoclonal antibody. In this patient group, the antibody 16H5 of the IgG1 isotype had been administered in a median total dosage of 140 mg per treatment cycle. Four patients took part in a second treatment regimen 6-8 weeks later. After the first treatment cycle, detectable HAMAs developed in 5 out of 10 patients. In 4 individuals undergoing a second course of therapy, increases of HAMAs were evident only in the 3 patients with previous HAMA responses. HAMAs were primarily of the IgG isotype, while the presence of rheumatoid factors usually interfered with the detectability of IgM HAMAs. However, using isolated F(ab)2 fragments of the monoclonal reagent used for therapy, HAMAs of the IgM isotype were also detectable. HAMAs of the IgG isotype did not exceed levels of 2.0 mg/liter after a single treatment cycle and 2.2 mg/liter after a repeated cycle. No IgE responses were detectable. Absorption experiments indicated that approximately 25% of the HAMA activity was directed against specific determinants of the 16H5 monoclonal antibody, presumably including anti-idiotypic reactivities. These data demonstrate that HAMAs developed only in a proportion of RA patients treated with the anti-CD4 monoclonal antibody 16H5. However, the amounts were rather low compared to other monoclonal reagents used in cancer patients and were therefore allowed for repeated applications without an apparent loss of efficacy.

  19. Sandwich ELISA Using a Mouse/Human Chimeric CSLEX-1 Antibody.

    Science.gov (United States)

    Yamashita, J; Kobayashi, I; Tatematsu, K; Sezutsu, H; Noda, K; Ishihara, H

    2016-11-01

    An assay using a mouse antisialyl Lewis X (sLeX) antibody (CSLEX-1) is used clinically for screening and monitoring patients with breast cancer in Japan. However, the IgM isoform of CSLEX-1 is not preferred for the assay because the bulkiness of IgM generally causes poor accessibility to the antigen. To solve this problem, we developed an antisLeX mouse/human chimeric IgG antibody, CH-CSLEX-1, using transgenic silkworms. The performance of a homologous sandwich ELISA of CH-CSLEX1 was then evaluated. To generate CH-CSLEX-1, we used a GAL4/UAS binary gene expression system in transgenic silkworms. The reactivities of CSLEX-1 and CH-CSLEX-1 were determined in a Biacore analysis. To confirm antigen specificity, 3 antigens [sLeX, sLeA, and Lewis Y (LeY)] were used. CH-CSLEX-1 formed correctly as an IgG class of immunoglobulin molecule with an isoelectric point close to the predicted value. The best combination for capturing and probing in a sandwich ELISA was determined as a homologous combination of CH-CSLEX-1. The CH-CSLEX-1 assay specifically detected sLeX, but not sLeA and LeY. A correlation analysis with 107 human samples showed good concordance between the conventional CSLEX-1 assay (homologous sandwich ELISA using CSLEX-1) and the CH-CSLEX-1 assay (r = 0.98). Moreover, the CH-CSLEX-1 assay was not affected by either human antimouse IgG antibodies (HAMA IgG) or HAMA IgM. The mouse/human chimeric antibody CH-CSLEX-1 allowed the establishment of a highly specific sandwich ELISA for sLeX that was not affected by HAMA. © 2016 American Association for Clinical Chemistry.

  20. Evaluation of six immunoassays for detection of dengue virus-specific immunoglobulin M and G antibodies

    NARCIS (Netherlands)

    J. Groen (Jan); P. Koraka (Penelope); J. Velzing (Jans); C. Copra (Cederick); A.D.M.E. Osterhaus (Albert)

    2000-01-01

    textabstractThe performance of six commercially available immunoassay systems for the detection of dengue virus-specific immunoglobulin M (IgM) and IgG antibodies in serum was evaluated. These included two IgM and IgG enzyme immunoassays (EIA) from MRL Laboratories and PanBio, a rapid

  1. The Epoch of IGM heating by early sources of X-rays

    Science.gov (United States)

    Eide, Marius B.; Graziani, Luca; Ciardi, Benedetta; Feng, Yu; Kakiichi, Koki; Di Matteo, Tiziana

    2018-02-01

    Observations of the 21 cm line from neutral hydrogen indicate that an Epoch of Heating (EoH) might have preceded the later Epoch of Reionization (EoR). Here we study the effects on the ionization state and the thermal history of the Intergalactic Medium (IGM) during the EoH induced by different assumptions on ionizing sources in the high redshift Universe: (i) stars, (ii) X-ray binaries (XRBs), (iii) thermal bremsstrahlung of the hot Interstellar Medium (ISM), and (iv) accreting nuclear black holes (BHs). To this aim, we post-process outputs from the (100 h-1 cMpc)3 hydrodynamical simulation MassiveBlack-II with the cosmological 3D radiative transfer code CRASH, which follows the propagation of UV and X-ray photons, computing the thermal and ionization state of hydrogen and helium through the EoH. We find that stars determine the fully ionized morphology of the IGM, while the spectrally hard XRBs pave way for efficient subsequent heating and ionization by the spectrally softer ISM. With the seeding prescription in MassiveBlack-II, BHs do not contribute significantly to either ionization or heating. With only stars, most of the IGM remains in a cold state (with a median T = 11 K at z = 10), however, the presence of more energetic sources raises the temperature of regions around the brightest and more clustered sources above that of the CMB, opening the possibility to observing the 21 cm signal in emission.

  2. The importance of IgM positivity in laboratory diagnosis of gestational and congenital syphilis.

    Science.gov (United States)

    Nemes-Nikodém, E; Vörös, E; Pónyai, K; Párducz, L; Kárpáti, S; Rozgonyi, F; Ostorházi, E

    2012-06-01

    From January 1, 2009 through December 31, 2011, from 33,753 blood samples for syphilis screening, Treponema pallidum infections were confirmed in 241 pregnant women at the Department of Dermatology, Venerology, and Dermatooncology of Semmelweis University Budapest. In this period, four children born to inadequately or untreated women were confirmed to have connatal syphilis. The height of rapid plasma reagin (RPR) titer was measured to determine the stage of the infection and to examine the success of the antilues therapy. The diagnosis of maternal syphilis infection was confirmed with enzyme linked immunosorbent assay (ELISA), T. pallidum particle agglutination (TPPA), and IgG and IgM immunoblots. Maternal IgM immunoblot results identify mothers at risk of delivering babies with connatal syphilis better than the height of maternal RPR titer. The standard serological tests are less useful in newborns because of IgG transfer across the placenta. IgM test which depends on the infant's response has more specificity in diagnosing connatal syphilis.

  3. A Cross Section Study to Determine the Prevalence of Antibodies against HIV Infection among Hepatitis B and C Infected Individuals

    Directory of Open Access Journals (Sweden)

    Geane L. Flores

    2016-03-01

    Full Text Available (1 Background: There are limited data regarding human immunodeficiency virus (HIV prevalence among hepatitis B virus (HBV or hepatitis C virus (HCV infected individuals. The aim of this cross-sectional study is to determine the prevalence of HBV and HCV infection among HIV individuals; (2 Methods: A total of 409 patients (126 HBV+ and 283 HCV+ referred to the Brazilian Reference Laboratory for Viral Hepatitis from 2010 to 2013 donated serum samples. Anti-HIV, HBsAg, anti-HBc, anti-HBs, anti-HBcIgM, anti-HBe, HBeAg, and anti-HCV antibodies were measured, and anti-HCV positive samples were tested for viral RNA and genotype; (3 Results: The anti-HIV antibody prevalence was 10.31% and 4.59% among HBV+ and HCV+ patients, respectively. The HCV mean (SD viral load was log 5.14 ± 1.64 IU/mL, and genotype I was most prevalent (163/283. Anti-HBs and anti-HBc were detected in 40% and 26% of HCV+ individuals, respectively. Among the HBV+ population, the presence of anti-HIV antibodies was associated with male gender, marital status (married, tattoo, sexual orientation, sexual practices (oral sex and anal sex, history of sexually transmitted diseases (STDs, history of viral hepatitis treatment, and a sexual partner with hepatitis or HIV. For the HCV+ group, the presence of anti-HIV antibodies was associated with female gender, marital status (married, anal intercourse, previous history of STDs, and number of sexual partners; (4 Conclusion: A high prevalence of anti-HIV antibodies was found among individuals with HBV and HCV, showing the importance of education programmes towards HIV infection among HBV- and HCV-infected individuals.

  4. Seroprevalence of Rubella antibodies among pregnant women with bad obstetric history in tertiary care hospital

    Directory of Open Access Journals (Sweden)

    Shashi Chopra

    2015-01-01

    Full Text Available Background Infection with rubella virus can be disastrous in early gestation. Rubella is a major cause of birth defects in infants and the risk of having congenital rubella in seronegative pregnant women is more in developing countries. Aim This study was carried to determine the seroprevalence of rubella antibodies in pregnant women. Setting and Design Three years retrospective study in the department of Microbiology, PIMS, Jalandhar. Material and method This study comprised of 250 pregnant women, whose blood samples were collected, sera were screened for rubella specific IgM and IgG antibodies by RecomLine ToRCH Result Out of 250 samples (Study group, 53% (n=132 were seropositive for IgG antibodies and 4.5% (n=9 were seropositive for IgM antibodies. Conclusion Our study demonstrates a strong association between rubella infection and BOH in women. Serosurveillance of Rubella among adolescent girls and women of child bearing age should be considered at a national level to prevent innumerable abortions, stillbirth and congenital anomalies due to rubella.

  5. Detection of Antibodies against Human and Plant Aquaporins in Patients with Multiple Sclerosis

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    Aristo Vojdani

    2015-01-01

    Full Text Available Multiple sclerosis (MS is an autoimmune disease that affects the body’s central nervous system. Around 90% of MS sufferers are diagnosed with relapsing-remitting MS (RRMS. We used ELISA to measure IgG, IgA, and IgM antibodies against linear epitopes of human and plant aquaporins (AQP4 as well as neural antigens in RRMS patients and controls to determine whether patients suffering from RRMS have simultaneous elevations in antibodies against these peptides and antigens. In comparison to controls, significant elevations in isotype-specific antibodies against human and plant AQP4 and neural antigens such as MBP, MOG, and S100B were detected in RRMS patients, indicating a high correlation in antibody reaction between plant aquaporins and brain antigens. This correlation between the reactivities of RRMS patients with various tested antigens was the most significant for the IgM isotype. We conclude that a subclass of patients with RRMS reacts to both plant and human AQP4 peptides. This immune reaction against different plant aquaporins may help in the development of dietary modifications for patients with MS and other neuroimmune disorders.

  6. Using Monoclonal Antibody to Determine Lead Ions with a Localized Surface Plasmon Resonance Fiber-optic Biosensor

    Directory of Open Access Journals (Sweden)

    Mon-Fu Chung

    2008-01-01

    Full Text Available A novel reflection-based localized surface plasmon resonance (LSPR fiber-optic probe has been developed to determine the heavy metal lead ion concentration. Monoclonal antibody as the detecting probe containing massive amino groups to capture Pb(II-chelate complexes was immobilized onto gold nanoparticle-modified optical fiber (NMAuOF. The optimal immobilizing conditions of monoclonal antibody on to the NMAuOF are 189 μg/mL in pH7.4 PBS for 2 h at 25°C. The absorbability of the functionalized NMAuOF sensor increases to 12.2 % upon changing the Pb(II-EDTA level from 10 to 100 ppb with a detection limit of 0.27 ppb. The sensor retains 92.7 % of its original activity and gives reproducible results after storage in 5% D-( -Trehalose dehydrate solution at 4°C for 35 days. In conclusion, the monoclonal antibody-functionalized NMAuOF sensor shows a promising result for determining the concentration of Pb(II with high sensitivity.

  7. Molecular Determinants for Antibody Binding on Group 1 House Dust Mite Allergens

    Energy Technology Data Exchange (ETDEWEB)

    Chruszcz, Maksymilian; Pomés, Anna; Glesner, Jill; Vailes, Lisa D.; Osinski, Tomasz; Porebski, Przemyslaw J.; Majorek, Karolina A.; Heymann, Peter W.; Platts-Mills, Thomas A.E.; Minor, Wladek; Chapman, Martin D. (INDOOR Bio.); (UV); (UVHS)

    2012-07-11

    House dust mites produce potent allergens, Der p 1 and Der f 1, that cause allergic sensitization and asthma. Der p 1 and Der f 1 are cysteine proteases that elicit IgE responses in 80% of mite-allergic subjects and have proinflammatory properties. Their antigenic structure is unknown. Here, we present crystal structures of natural Der p 1 and Der f 1 in complex with a monoclonal antibody, 4C1, which binds to a unique cross-reactive epitope on both allergens associated with IgE recognition. The 4C1 epitope is formed by almost identical amino acid sequences and contact residues. Mutations of the contact residues abrogate mAb 4C1 binding and reduce IgE antibody binding. These surface-exposed residues are molecular targets that can be exploited for development of recombinant allergen vaccines.

  8. Circulating microparticles in patients with antiphospholipid antibodies: characterization and associations.

    Science.gov (United States)

    Chaturvedi, Shruti; Cockrell, Erin; Espinola, Ricardo; Hsi, Linda; Fulton, Stacey; Khan, Mohammad; Li, Liang; Fonseca, Fabio; Kundu, Suman; McCrae, Keith R

    2015-01-01

    The antiphospholipid syndrome is characterized by venous or arterial thrombosis and/or recurrent fetal loss in the presence of circulating antiphospholipid antibodies. These antibodies cause activation of endothelial and other cell types leading to the release of microparticles with procoagulant and pro-inflammatory properties. The aims of this study were to characterize the levels of endothelial cell, monocyte or platelet derived, and tissue factor-bearing microparticles in patients with antiphospholipid antibodies, to determine the association of circulating microparticles with anticardiolipin and anti-β2-glycoprotein antibodies, and to define the cellular origin of microparticles that express tissue factor. Microparticle content within citrated blood from 47 patients with antiphospholipid antibodies and 144 healthy controls was analyzed within 2hours of venipuncture. Levels of Annexin-V, CD105 and CD144 (endothelial derived), CD41 (platelet derived) and tissue factor positive microparticles were significantly higher in patients than controls. Though levels of CD14 (monocyte-derived) microparticles in patient plasma were not significantly increased, increased levels of CD14 and tissue factor positive microparticles were observed in patients. Levels of microparticles that stained for CD105 and CD144 showed a positive correlation with IgG (R=0.60, p=0.006) and IgM anti-beta2-glycoprotein I antibodies (R=0.58, p=0.006). The elevation of endothelial and platelet derived microparticles in patients with antiphospholipid antibodies and their correlation with anti-β2-glycoprotein I antibodies suggests a chronic state of vascular cell activation in these individuals and an important role for β2-glycoprotein I in development of the pro-thrombotic state associated with antiphospholipid antibodies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

    OpenAIRE

    Yang, Danlin; Singh, Ajit; Wu, Helen; Kroe-Barrett, Rachel

    2017-01-01

    Label-free optical biosensors are powerful tools in drug discovery for the characterization of biomolecular interactions. In this study, we describe the use of four routinely used biosensor platforms in our laboratory to evaluate the binding affinity and kinetics of ten high-affinity monoclonal antibodies (mAbs) against human proprotein convertase subtilisin kexin type 9 (PCSK9). While both Biacore T100 and ProteOn XPR36 are derived from the well-established Surface Plasmon Resonance (SPR) te...

  10. Determining vaccination frequency in farmed rainbow trout using Vibrio anguillarum O1 specific serum antibody measurements.

    Directory of Open Access Journals (Sweden)

    Lars Holten-Andersen

    Full Text Available BACKGROUND: Despite vaccination with a commercial vaccine with a documented protective effect against Vibrio anguillarum O1 disease outbreaks caused by this bacterium have been registered among rainbow trout at Danish fish farms. The present study examined specific serum antibody levels as a valid marker for assessing vaccination status in a fish population. For this purpose a highly sensitive enzyme-linked immunosorbent assay (ELISA was developed and used to evaluate sera from farmed rainbow trout vaccinated against V. anguillarum O1. STUDY DESIGN: Immune sera from rainbow trout immunised with an experimental vaccine based on inactivated V. anguillarum O1 bacterin in Freund's incomplete adjuvant were used for ELISA optimisation. Subsequently, sera from farmed rainbow trout vaccinated with a commercial vaccine against V. anguillarum were analysed with the ELISA. The measured serum antibody levels were compared with the vaccine status of the fish (vaccinated/unvaccinated as evaluated through visual examination. RESULTS: Repeated immunisation with the experimental vaccine lead to increasing levels of specific serum antibodies in the vaccinated rainbow trout. The farmed rainbow trout responded with high antibody levels to a single injection with the commercial vaccine. However, the diversity in responses was more pronounced in the farmed fish. Primary visual examinations for vaccine status in rainbow trout from the commercial farm revealed a large pool of unvaccinated specimens (vaccination failure rate=20% among the otherwise vaccinated fish. Through serum analyses using the ELISA in a blinded set-up it was possible to separate samples collected from the farmed rainbow trout into vaccinated and unvaccinated fish. CONCLUSIONS: Much attention has been devoted to development of new and more effective vaccines. Here we present a case from a Danish rainbow trout farm indicating that attention should also be directed to the vaccination procedure in

  11. Determination of specific antibodies titre to salmonella enteritidis by elisa technique in several selected flocks of laying hens

    Directory of Open Access Journals (Sweden)

    Velhner Maja

    2004-01-01

    Full Text Available In this paper, the antibody titre to Salmonella enteritidis (SE was examined by the ELISA method in two flocks of laying hens, where during routine bacteriological investigations Salmonellae was never isolated, and in one flock where Colysepticemia was diagnosed and Salmonella isolated accidentally. In the flocks were Salmonellae were not isolated, a titre with a high level of specific antibodies to SE was discovered (15 and 45%, while the flock with accidental findings of SE was poorly positive (5%. These results point to the necessity of introducing serological monitoring to SE so that the infection of salmonella may be discovered early and the prevalence in the flock determined, and also for the purpose of applying adequate measures that could reduce the possibility of secretion of SE through eggs.

  12. Persistence of the protective immunity and kinetics of the isotype specific antibody response against the viral nucleocapsid protein after experimental Schmallenberg virus infection of sheep.

    Science.gov (United States)

    Poskin, Antoine; Verite, Stephanie; Comtet, Loic; Van der Stede, Yves; Cay, Brigitte; De Regge, Nick

    2015-10-15

    Schmallenberg virus (SBV) is an Orthobunyavirus that induces abortion, stillbirths and congenital malformations in ruminants. SBV infection induces a long lasting seroconversion under natural conditions. The persistence of the protective immunity and the isotype specific antibody response upon SBV infection of sheep has however not been studied in detail. Five sheep were kept in BSL3 facilities for more than 16 months and subjected to repeated SBV infections. Blood was regularly sampled and organs were collected at euthanasia. The presence of SBV RNA in serum and organs was measured with quantitative real-time PCR. The appearance and persistence of neutralizing and SBV nucleoprotein (N) isotype specific antibodies was determined with virus neutralization tests (VNT) and ELISAs. The primo SBV infection protected ewes against clinical signs, viraemia and virus replication in organs upon challenge infections more than 15 months later. Production of neutralizing SBV specific antibodies was first detected around 6 days post primo-inoculation with VNT and correlated with the appearance of SBV-N specific IgM antibodies. These IgM antibodies remained present for 2 weeks. SBV-N specific IgG antibodies were first detected between 10 and 21 dpi and reached a plateau at 28 dpi. This plateau remained consistently high and no significant decrease in titre was found over a period of more than 1 year. Similar results were found for the neutralising antibody response. In conclusion, the SBV specific IgM response probably eliminates SBV from the blood and the protective immunity induced by SBV infection protects sheep against reinfection for at least 16 months.

  13. Deterioration of lung function is associated with presence of IgM rheumatoid factor and smoking in patients with systemic sclerosis

    DEFF Research Database (Denmark)

    Broholm, B.; Ullman, S.; Halberg, P.

    2008-01-01

    of patients with systemic sclerosis (SSc) the influence of smoking and IgM RF on the lung function was calculated. One hundred fifty-five persons with SSc had vital capacity (VC) and diffusing capacity (DLco) measured at least twice with at least 1-year interval as percents of predicted values according...... and whether the presence of IgM RF in smoking patients with SSc actually confers an increased risk of pulmonary damage remains to be determined Udgivelsesdato: 2008/8......Smoking is a known risk factor for the development of several lung diseases, autoimmune diseases, and IgM rheumatoid factor (RF) in nonrheumatic persons. In patients with rheumatoid arthritis and IgM RF the diffusion capacity is decreased in smokers but not in nonsmokers. In the present study...

  14. Deterioration of lung function is associated with presence of IgM rheumatoid factor and smoking in patients with systemic sclerosis

    DEFF Research Database (Denmark)

    Broholm, Berit; Ullman, Susanne; Halberg, Poul

    2008-01-01

    of patients with systemic sclerosis (SSc) the influence of smoking and IgM RF on the lung function was calculated. One hundred fifty-five persons with SSc had vital capacity (VC) and diffusing capacity (DLco) measured at least twice with at least 1-year interval as percents of predicted values according...... and whether the presence of IgM RF in smoking patients with SSc actually confers an increased risk of pulmonary damage remains to be determined.......Smoking is a known risk factor for the development of several lung diseases, autoimmune diseases, and IgM rheumatoid factor (RF) in nonrheumatic persons. In patients with rheumatoid arthritis and IgM RF the diffusion capacity is decreased in smokers but not in nonsmokers. In the present study...

  15. Determination of specificity and pattern of antinuclear antibodies (ana) in systemic rheumatic disease patients positive for ana testing

    International Nuclear Information System (INIS)

    Nawaz, H.; Bashir, M.M.; Iqbal, W.

    2018-01-01

    To determine probability of finding antinuclear antibodies (ANA) and anti extractable nuclear antigens (ENA) positive samples and associating ANA patterns with anti-ENA reactivities among a consecutive cohort of samples of systemic rheumatic disease patients referred for ANA testing. Study Design:Prospective cohort study. Place and Duration of Study:Immunology Department, Armed Forces Institute of Pathology, Rawalpindi, Pakistan, from January to June 2016. Methodology:All the samples referred for ANA testing with clinical suspicion of systemic rheumatic disease were included. After screening, ANA positive samples were subjected to anti-ENA antibodies testing (including anti-SSA, anti-SSB, anti-Sm, anti-RNP, anti-SCL-70 and anti-Jo-1 antibodies) and ANA pattern and titer determination. Results:Of 4,347 samples received, 397 were positive for ANA (9%). Of 397, 96 (24%) samples positive on ENA screen were tested for anti-ENA reactivity. Anti-SSA antibodies were found in 59 samples. Commonest ANA patterns were coarse and fine speckled (43 and 22 samples of 81 tested), while majority of samples carried ANA in titers of 1:40 and 1:80 (22 and 18 samples of 81 tested). No specific ANA pattern was associated with any particular anti-ENA reactivity. Conclusion:Among samples/patients referred for investigations of autoimmune disorders, probability of finding positive ANA is approximately 9%. Of these 9%, about 24% also show reactivity against ENA. Commonest ANA pattern is coarse speckled and majority of such patients carry ANA in titers ranging from 1:40 to 1:80. Commonest ENA reactivity was against SSA. (author)

  16. Anti-ganglioside antibodies in amyotrophic lateral sclerosis revisited.

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    Katja Kollewe

    Full Text Available Amyotrophic Lateral Sclerosis (ALS is a devastating neurodegenerative disorder with typical onset in the 5th- 6th decade of life. The hypothesis of an autoimmune origin of ALS receives less attention today, but immunological phenomena still seem to be involved and mechanisms such as protective autoimmunity may be important. Detection of antibodies against a variety of gangliosides has been repeatedly described in ALS-patients by several authors, but widely differing frequencies and titres have been reported. Therefore, we investigated the presence of six common antibodies with a commercially available test panel for GA1, GM1, GM2, GD1a, GD1b and GQ1b in a large group of clinically well-characterized ALS patients and compared them to a collective of 200 healthy blood donors.IgG and IgM antibodies to the six gangliosides asialoGM1 (GA1, GM1, GM2, GD1a, GD1b, GQ1b were determined by GanglioCombi ELISA in sera of 84 ALS patients. Results were expressed as a %-ratio of a highly positive control and categorized as negative (100%. The values obtained from 200 Swiss blood donors served as a reference group.In twenty-two (26.2% ALS-patients elevated anti-ganglioside antibodies could be detected: Taking all subspecific antibodies together, IgG antibodies were found in 9/84 (10.7% and IgM in 15/84 (17.9% patients. There was no correlation between age, gender, site of onset or survival and anti-ganglioside-positive/-negative titres in ALS-patients. No statistically significant difference in the frequency of anti-ganglioside antibodies compared to the group of healthy blood donors was found.Even with this more comprehensive approach, anti-ganglioside antibody frequencies and patterns in our ALS cohort closely resembled the values measured in healthy controls. In accordance with other studies, we did not observe any association of a distinct ALS phenotype with elevated anti-ganglioside antibodies or an impact on survival.

  17. Utility of IgM ELISA, TaqMan real-time PCR, reverse transcription PCR, and RT-LAMP assay for the diagnosis of Chikungunya fever.

    Science.gov (United States)

    Reddy, Vijayalakshmi; Ravi, Vasanthapuram; Desai, Anita; Parida, Manmohan; Powers, Ann M; Johnson, Barbara W

    2012-11-01

    Chikungunya fever a re-emerging infection with expanding geographical boundaries, can mimic symptoms of other infections like dengue, malaria which makes the definitive diagnosis of the infection important. The present study compares the utility of four laboratory diagnostic methods viz. IgM capture ELISA, an in house reverse transcription PCR for the diagnosis of Chikungunya fever, TaqMan real-time PCR, and a one step reverse transcription-loop mediated isothermal amplification assay (RT-LAMP). Out of the 70 serum samples tested, 29 (41%) were positive for Chikungunya IgM antibody by ELISA and 50 (71%) samples were positive by one of the three molecular assays. CHIKV specific nucleic acid was detected in 33/70 (47%) by reverse transcription PCR, 46/70 (66%) by TaqMan real-time PCR, and 43/70 (62%) by RT-LAMP assay. A majority of the samples (62/70; 89%) were positive by at least one of the four assays used in the study. The molecular assays were more sensitive for diagnosis in the early stages of illness (2-5 days post onset) when antibodies were not detectable. In the later stages of illness, the IgM ELISA is a more sensitive diagnostic test. In conclusion we recommend that the IgM ELISA be used as an initial screening test followed one of the molecular assays in samples that are collected in the early phase of illness and negative for CHIKV IgM antibodies. Such as approach would enable rapid confirmation of the diagnosis and implementation of public health measures especially during outbreaks. Copyright © 2012 Wiley Periodicals, Inc.

  18. Comparison of Seven Commercial Antigen and Antibody Enzyme-Linked Immunosorbent Assays for Detection of Acute Dengue Infection

    Science.gov (United States)

    Jarman, Richard G.; Gibbons, Robert V.; Tanganuchitcharnchai, Ampai; Mammen, Mammen P.; Nisalak, Ananda; Kalayanarooj, Siripen; Bailey, Mark S.; Premaratna, Ranjan; de Silva, H. Janaka; Day, Nicholas P. J.; Lalloo, David G.

    2012-01-01

    Seven commercial assays were evaluated to determine their suitability for the diagnosis of acute dengue infection: (i) the Panbio dengue virus Pan-E NS1 early enzyme-linked immunosorbent assay (ELISA), second generation (Alere, Australia); (ii) the Panbio dengue virus IgM capture ELISA (Alere, Australia); (iii) the Panbio dengue virus IgG capture ELISA (Alere, Australia); (iv) the Standard Diagnostics dengue virus NS1 antigen ELISA (Standard Diagnostics, South Korea); (v) the Standard Diagnostics dengue virus IgM ELISA (Standard Diagnostics, South Korea); (vi) the Standard Diagnostics dengue virus IgG ELISA (Standard Diagnostics, South Korea); and (vii) the Platelia NS1 antigen ELISA (Bio-Rad, France). Samples from 239 Thai patients confirmed to be dengue virus positive and 98 Sri Lankan patients negative for dengue virus infection were tested. The sensitivities and specificities of the NS1 antigen ELISAs ranged from 45 to 57% and 93 to 100% and those of the IgM antibody ELISAs ranged from 85 to 89% and 88 to 100%, respectively. Combining the NS1 antigen and IgM antibody results from the Standard Diagnostics ELISAs gave the best compromise between sensitivity and specificity (87 and 96%, respectively), as well as providing the best sensitivity for patients presenting at different times after fever onset. The Panbio IgG capture ELISA correctly classified 67% of secondary dengue infection cases. This study provides strong evidence of the value of combining dengue virus antigen- and antibody-based test results in the ELISA format for the diagnosis of acute dengue infection. PMID:22441389

  19. Antiphospholipid Antibody Titers and Clinical Outcomes in Patients with Recurrent Miscarriage and Antiphospholipid Antibody Syndrome: A Prospective Study

    Directory of Open Access Journals (Sweden)

    Yu Song

    2017-01-01

    Conclusions: Anti-β2-GP1 IgM was the predominant form of antibody in patients with RM and APS. The decreases in antiphospholipid antibody titers correlated with better pregnancy outcomes. The shorter treatment regimen was effective and economical.

  20. Evaluation of a radioimmunoassay for the determination of anti-native DNA antibodies

    International Nuclear Information System (INIS)

    Johanet, C.; Soulie, E.; Absalon, Y.B.; Ocwieja, T.; Abuaf, N.

    1991-01-01

    The anti-native DNA antibodies were measured by a radioimmunoassay (RIA) type Farr assay in the sera from 648 patients: 108 with active or inactive systemic lupus erythematosus (SLE), 181 with clinical symptoms of another connective tissue disease, 171 with liver diseases, 29 with different pathology and 159 normal sera were obtained from a blood bank. The anti-DNA kit has been calibrated against the first international units/ml. This assay has proved to be sensitive and specific, and appears to be reliable for the diagnosis and follow-up of SLE patients. The authors propose a new reference cut-off level higher than producer's one [fr

  1. Radioimmunoassay of measles virus antibodies in SSPE

    International Nuclear Information System (INIS)

    Jankowski, M.A.; Gut, W.; Kantoch, M.

    1982-01-01

    A sensitive radioimmunoassay (RIA) was introduced for detecting measles virus IgG and IgM antibodies. The hyperimmune response to the measles virus could be demonstrated more accurately by RIA than by haemagglutination inhibition (HI). The ratio between RIA and HI antibody titres was decidedly higher in sera and cerebrospinal fluids of patients with subacute sclerosing panencephalitis than in those of other groups tested. (author)

  2. [Detection of cerebrospinal fluid anti-enterovirus 71 IgM in children with severe hand, food and mouth disease induced by enterovirus 71 infection and its clinical significance].

    Science.gov (United States)

    Wu, Yidong; Chen, Dong; Zhou, Jun; Wang, Jie; Zhao, Shiyong; Shao, Qimin; Wei, Yi

    2015-05-01

    To detect the anti-enterovirus 71 (EV71) IgM level in cerebrospinal fluid (CSF) of children with severe hand, foot and mouth disease (HFMD) induced by EV71 and then analyze the relationships among the IgM antibody levels, CSF routine examination and patients' clinical features, and thus to evaluate the clinical significance of anti-EV71 IgM as a new indicator for early diagnosis of children with severe HFMD induced by EV71. A total of 294 laboratory-confirmed cases of children with severe HFMD infected with EV71 were enrolled into the research group from March 2014 to June 2014, consisting of 53 fatal cases and 241 severe cases, and their CSF samples underwent enzyme-linked immunosorbent assay (ELISA) for anti-EV71 IgM levels, CSF routine and biochemical tests. Forty-one cases of children with severe HFMD induced by other enteroviruses were collected as antibody-testing control group during the same period. In the research group, the total positive rate of anti-EV71 IgM in 294 CSF samples of children with severe HFMD infected by EV71 was 60.2% (177/294); the positive rate of anti-EV71 IgM in the fatal HFMD subgroup was 62.3% (33/53); the positive rate of anti-EV71 IgM in the severe HFMD subgroup was 59.8% (144/241). In the control group, the results of CSF anti-EV71 IgM tests were all negative (0/41). In the research group, patients in antibody-positive subgroup (2.5±1.2) years old were younger than those in antibody-negative subgroup (2.9±1.1) years old (t=2.595, P=0.010). And within the antibody-positive subgroup, the patients ((1.9±0.7) years old) with fatal type disease were younger than those ((2.6±1.2) years old) with severe type disease (t=3.150, P=0.002). The CSF nucleated cells count and positive rates (105 (56,180) ×10(6) /L; 97.7% (173/177)) in antibody-positive subgroup were higher than those (62(30,150) ×10(6) /L; 83.8% (98/117)) in antibody-negative subgroup (Z=3.663, P=0.000; χ(2)=19.089, P=0.000). In antibody-positive subgroup, the percentage

  3. Determinants of tick-borne encephalitis virus antibody presence in roe deer (Capreolus capreolus) sera.

    Science.gov (United States)

    Kiffner, C; Vor, T; Hagedorn, P; Niedrig, M; Rühe, F

    2012-03-01

    In order to identify variables associated with the presence of the tick-borne encephalitis (TBE) virus, we conducted a serological survey of roe deer [Capreolus capreolus (Artiodactyla: Cervidae, Linnaeus 1758)] in three forest districts of southern Hesse, Germany. Overall, 24 out of 105 (22.9%) of the sera were positive (≥1 : 10 plaque reduction neutralization test). Using a logistic regression approach, we found that unexplained spatial variation, indexed roe deer density (positive correlation), hind foot length of the tested roe deer (positive correlation) and infestation with female Ixodes spp. ticks (negative correlation) predicted the probability of TBE virus antibody presence in individual roe deer sera. Spring temperature increase and host sex were rejected as explanatory variables. We found considerable differences in TBE virus antibody seroprevalence (50.0% vs. 17.6%) between two forest districts located in the same county; this finding questions the current county-resolution of public health recordings. Given the high seroprevalence of roe deer and the considerable explanatory power of our model, our approach appears suitable to delineate science-based risk maps at a smaller spatial scale and to abandon the current human incidence per county criterion. Importantly, using roe deer as sentinels would eliminate the inherent bias of risk maps based on human incidence (varying levels of immunization and exposure of humans). © 2011 The Authors. Medical and Veterinary Entomology © 2011 The Royal Entomological Society.

  4. Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA.

    Science.gov (United States)

    Wang, Ting; Li, Peiwu; Zhang, Qi; Zhang, Wen; Zhang, Zhaowei; Wang, Tong; He, Ting

    2017-06-28

    Aspergillus and its poisonous mycotoxins are distributed worldwide throughout the environment and are of particular interest in agriculture and food safety. In order to develop a specific method for rapid detection of Aspergillus flavus to forecast diseases and control aflatoxins, a nanobody, PO8-VHH, highly reactive to A. flavus was isolated from an immunized alpaca nanobody library by phage display. The nanobody was verified to bind to the components of extracellular and intracellular antigen from both A. flavus and A. parasiticus. To construct a sandwich format immunoassay, polyclonal antibodies against Aspergillus were raised with rabbits. Finally, a highly selective nanobody-polyclonal antibody sandwich enzyme-linked immunosorbent assay was optimized and developed. The results revealed that the detection limits of the two fungi were as low as 1 μg mL -1 , and that it is able to detect fungal concentrations below to 2 μg mg -1 of peanut and maize grains in both artificially and naturally contaminated samples. Therefore, we here provided a rapid and simple method for monitoring Aspergillus spp. contamination in agricultural products.

  5. Antiphospholipid antibodies in Brazilian hepatitis C virus carriers

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    A.M. Atta

    2008-06-01

    Full Text Available Hepatitis C, a worldwide viral infection, is an important health problem in Brazil. The virus causes chronic infection, provoking B lymphocyte dysfunction, as represented by cryoglobulinemia, non-organ-specific autoantibody production, and non-Hodgkin's lymphoma. The aim of this research was to screen for the presence of antiphospholipid autoantibodies in 109 Brazilian hepatitis C virus carriers without clinical history of antiphospholipid syndrome. Forty healthy individuals were used as the control group. IgA, IgG, and IgM antibodies against cardiolipin and β2-glycoprotein I were measured with an enzyme-linked immunosorbent assay, using a cut-off point of either 20 UPL or 20 SBU. While 24 (22.0% hepatitis C carriers had moderate titers of IgM anticardiolipin antibodies (median, 22.5 MPL; 95%CI: 21.5-25.4 MPL, only three carriers (<3% had IgG anticardiolipin antibodies (median, 23 GPL; 95%CI: 20.5-25.5 GPL. Furthermore, IgA anticardiolipin antibodies were not detected in these individuals. Male gender and IgM anticardiolipin seropositivity were associated in the hepatitis C group (P = 0.0004. IgA anti-β2-glycoprotein-I antibodies were detected in 29 of 109 (27.0% hepatitis C carriers (median, 41 SAU; 95%CI: 52.7-103.9 SAU. Twenty patients (18.0% had IgM anti-β2-glycoprotein I antibodies (median, 27.6 SMU; 95%CI: 23.3-70.3 SMU, while two patients had IgG antibodies against this protein (titers, 33 and 78 SGU. Antiphospholipid antibodies were detected in only one healthy individual, who was seropositive for IgM anticardiolipin. We concluded that Brazilian individuals chronically infected with hepatitis C virus present a significant production of antiphospholipid antibodies, mainly IgA anti-β2-glycoprotein I antibodies, which are not associated with clinical manifestations of antiphospholipid syndrome.

  6. Possible role of specific immunoglobulin M antibodies to Plasmodium falciparum antigens in immunoprotection of humans living in a hyperendemic area, Burkina Faso

    DEFF Research Database (Denmark)

    Boudin, C; Chumpitazi, B; Dziegiel, M

    1993-01-01

    of antibodies to crude extracts of Plasmodium falciparum (IgG or IgM antisomatic and IgG antiexoantigens) were tested by IFI or enzyme-linked immunosorbent assay and were followed up according to the fluctuations of the parasite densities. Specific IgG antibodies had the same evolution as parasite densities....... Group 3 was composed of immunoprotected adults. Specific IgM and IgG antibodies to crude extracts or a recombinant antigen (glutamate-rich protein) of P. falciparum were tested. Specific IgM antibodies were lower in group 1 (nonimmune) than in groups 2 (semiimmune) and 3 (immunoprotected). Furthermore...

  7. Immunotherapy for IgM anti-myelin-associated glycoprotein paraprotein-associated peripheral neuropathies.

    Science.gov (United States)

    Lunn, Michael Pt; Nobile-Orazio, Eduardo

    2016-10-04

    Serum monoclonal anti-myelin-associated glycoprotein (anti-MAG) antibodies may be pathogenic in some people with immunoglobulin M (IgM) paraprotein and demyelinating neuropathy. Immunotherapies aimed at reducing the level of these antibodies might be expected to be beneficial. This is an update of a review first published in 2003 and previously updated in 2006 and 2012. To assess the effects of immunotherapy for IgM anti-MAG paraprotein-associated demyelinating peripheral neuropathy. On 1 February 2016 we searched the Cochrane Neuromuscular Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, and Embase for randomised controlled trials (RCTs). We also checked trials registers and bibliographies, and contacted authors and experts in the field. We included randomised controlled trials (RCTs) or quasi-RCTs involving participants of any age treated with any type of immunotherapy for anti-MAG antibody-associated demyelinating peripheral neuropathy with monoclonal gammopathy of undetermined significance and of any severity.Our primary outcome measures were numbers of participants improved in disability assessed with either or both of the Neuropathy Impairment Scale (NIS) or the modified Rankin Scale (mRS) at six months after randomisation. Secondary outcome measures were: mean improvement in disability, assessed with either the NIS or the mRS, 12 months after randomisation; change in impairment as measured by improvement in the 10-metre walk time, change in a validated linear disability measure such as the Rasch-built Overall Disability Scale (R-ODS) at six and 12 months after randomisation, change in subjective clinical scores and electrophysiological parameters at six and 12 months after randomisation; change in serum IgM paraprotein concentration or anti-MAG antibody titre at six months after randomisation; and adverse effects of treatments. We followed standard methodological procedures expected by Cochrane. We identified eight

  8. Antiphospholipid Antibodies in Lupus Nephritis.

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    Ioannis Parodis

    Full Text Available Lupus nephritis (LN is a major manifestation of systemic lupus erythematosus (SLE. It remains unclear whether antiphospholipid antibodies (aPL alter the course of LN. We thus investigated the impact of aPL on short-term and long-term renal outcomes in patients with LN. We assessed levels of aPL cross-sectionally in SLE patients diagnosed with (n = 204 or without (n = 294 LN, and prospectively in 64 patients with active biopsy-proven LN (52 proliferative, 12 membranous, before and after induction treatment (short-term outcomes. Long-term renal outcome in the prospective LN cohort was determined by the estimated glomerular filtration rate (eGFR and the Chronic Kidney Disease (CKD stage, after a median follow-up of 11.3 years (range: 3.3-18.8. Cross-sectional analysis revealed no association between LN and IgG/IgM anticardiolipin or anti-β2-glycoprotein I antibodies, or lupus anticoagulant. Both aPL positivity and levels were similar in patients with active LN and non-renal SLE. Following induction treatment for LN, serum IgG/IgM aPL levels decreased in responders (p<0.005 for all, but not in non-responders. Both at active LN and post-treatment, patients with IgG, but not IgM, aPL had higher creatinine levels compared with patients without IgG aPL. Neither aPL positivity nor levels were associated with changes in eGFR from either baseline or post-treatment through long-term follow-up. Moreover, aPL positivity and levels both at baseline and post-treatment were similar in patients with a CKD stage ≥3 versus 1-2 at the last follow-up. In conclusion, neither aPL positivity nor levels were found to be associated with the occurrence of LN in SLE patients. However, IgG aPL positivity in LN patients was associated with a short-term impairment of the renal function while no effect on long-term renal outcome was observed. Furthermore, IgG and IgM aPL levels decreased following induction treatment only in responders, indicating that aPL levels are

  9. CIRCULATING MICROPARTICLES IN PATIENTS WITH ANTIPHOSPHOLIPID ANTIBODIES: CHARACTERIZATION AND ASSOCIATIONS

    Science.gov (United States)

    Chaturvedi, Shruti; Cockrell, Erin; Espinola, Ricardo; Hsi, Linda; Fulton, Stacey; Khan, Mohammad; Li, Liang; Fonseca, Fabio; Kundu, Suman; McCrae, Keith R.

    2014-01-01

    The antiphospholipid syndrome is characterized by venous or arterial thrombosis and/or recurrent fetal loss in the presence of circulating antiphospholipid antibodies. These antibodies cause activation of endothelial and other cell types leading to the release of microparticles with procoagulant and pro-inflammatory properties. The aims of this study were to characterize the levels of endothelial cell, monocyte, platelet derived, and tissue factor-bearing microparticles in patients with antiphospholipid antibodies, to determine the association of circulating microparticles with anticardiolipin and anti-β2-glycoprotein antibodies, and to define the cellular origin of microparticles that express tissue factor. Microparticle content within citrated blood from 47 patients with antiphospholipid antibodies and 144 healthy controls was analyzed within 2 hours of venipuncture. Levels of Annexin-V, CD105 and CD144 (endothelial derived), CD41 (platelet derived) and tissue factor positive microparticles were significantly higher in patients than controls. Though levels of CD14 (monocyte-derived) microparticles in patient plasma were not significantly increased, increased levels of CD14 and tissue factor positive microparticles were observed in patients. Levels of microparticles that stained for CD105 and CD144 showed a positive correlation with IgG (R = 0.60, p=0.006) and IgM anti-beta2-glycoprotein I antibodies (R=0.58, p=0.006). The elevation of endothelial and platelet derived microparticles in patients with APS and their correlation with anti-β2-glycoprotein I antibodies suggests a chronic state of vascular cell activation in these individuals and an important role for β2-glycoprotein I in development of the pro-thrombotic state associated with antiphospholipid antibodies. PMID:25467081

  10. Kinetics of intralymphatically delivered monoclonal antibodies

    International Nuclear Information System (INIS)

    Wahl, R.L.; Geatti, O.; Liebert, M.; Beers, B.; Jackson, G.; Laino, L.; Kronberg, S.; Wilson, B.S.; Beierwaltes, W.H.

    1985-01-01

    Radiolabeled monoclonal antibody (MoAb) administration subcutaneously (sq), so that preferential uptake is to the lymphatics, holds significant promise for the detection of lymph node metastases. Only limited information is available about clearance rates of intralymphatically administered MoAbs. I-131 labeled intact IgG (225.28S), F(ab's)2 (225.28S) or IgM (FT162) were administered sq to anesthetized Balb/C mice. Eight mice were studied with each MoAb, 4 with a foot-pad injection, 4 with an anterior abdominal injection. Gamma camera images were collected into a computer, over the first 6 hrs after injection with the animals anesthetized and immobile. Animals were then allowed to move about freely. Additional images were then acquired out to 48 hrs. Regions of interest wre selected over the injection site and the kinetics of antibody egress determined. Clearance rates from local sq injection sites are influenced by motion and somewhat by location. The class and fragment status of the MoAb appear relatively less important in determining clearance rates from sq injections than they are in determining whole-body clearance after iv injections. Additional studies using Fab fragments and additional monoclonals will be useful in extending these observations

  11. Genetics Home Reference: X-linked hyper IgM syndrome

    Science.gov (United States)

    ... Conditions X-linked hyper IgM syndrome X-linked hyper IgM syndrome Printable PDF Open All Close All ... view the expand/collapse boxes. Description X-linked hyper IgM syndrome is a condition that affects the ...

  12. Incorporation of the ELISA technique to determine antibody levels against foot-and-mouth disease

    International Nuclear Information System (INIS)

    Vergara, N.N.; Caballero, P.H.; Santiago Gonzalez Patino, S.; Orue, P.M.

    1998-01-01

    Two groups of sera were evaluated by a liquid phase blocking ELISA (LPBE) for the detection and quantification of foot-and-mouth disease (FMD) antibodies to serotypes O, A and C to assess the sensitivity and specificity of the assay. The first group consisted of 120 sera from non-infected and non-vaccinated cattle, which were tested by a screening assay at a fix dilution of 1/32. The second group consisted of 120 sera from cattle vaccinated with a trivalent (O, A and C) vaccine. Sera from this group were titrated in a five fold dilution range: 1/10, 1/50, 1/250 and 1/1250. (author)

  13. Determination of Mother Centriole Maturation in CPAP-Depleted Cells Using the Ninein Antibody

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    Miseon Lee

    2015-03-01

    Full Text Available BackgroundMutations in centrosomal protein genes have been identified in a number of genetic diseases in brain development, including microcephaly. Centrosomal P4.1-associated protein (CPAP is one of the causal genes implicated in primary microcephaly. We previously proposed that CPAP is essential for mother centriole maturation during mitosis.MethodsWe immunostained CPAP-depleted cells using the ninein antibody, which selectively detects subdistal appendages in mature mother centrioles.ResultsNinein signals were significantly impaired in CPAP-depleted cells.ConclusionThe results suggest that CPAP is required for mother centriole maturation in mammalian cells. The selective absence of centriolar appendages in young mother centrioles may be responsible for asymmetric spindle pole formation in CPAP-depleted cells.

  14. Antiphosphatidylserine/prothrombin (aPS/PT) antibodies are associated with Raynaud phenomenon and migraine in primary thrombotic antiphospholipid syndrome.

    Science.gov (United States)

    Kopytek, M; Natorska, J; Undas, A

    2018-04-01

    Objectives Antibodies to phosphatidylserine/prothrombin complex (aPS/PT) detectable in sera of some patients with antiphospholipid syndrome (APS) have been shown to correlate with thrombosis. However, associations of aPS/PT antibodies with APS related disorders remain unclear. Aim To evaluate whether there are any associations between aPS/PT antibodies and Raynaud phenomenon, migraine and/or valvular lesions in primary thrombotic APS (PAPS). Methods We enrolled 67 consecutive patients (56 women) with thrombotic PAPS (VTE in 80.6%), aged 46.2 ± 13.5 years. The exclusion criteria were: acute coronary syndromes or stroke within preceding 6 months, cancer, severe comorbidities and pregnancy. The IgG and IgM aPS/PT antibodies were determined by ELISA with the cut-off of 30 units. We recorded Raynaud phenomenon, migraine and valvular lesions. Results Positive IgM or/and IgG aPS/PT antibodies were observed in 29 patients (43.3%), with a higher prevalence of IgM antibodies ( n = 27, 40.3%) compared with IgG isotype ( n = 12, 17.9%, p = 0.014). aPS/PT antibodies were observed most commonly in patients with triple aPL ( n = 12, 85.7%) compared with those with double ( n = 5, 35.7%) or single aPL antibodies (n = 12, 30.8%, p = 0.03), with no association with demographics, the ANA titre, the type of thrombotic events or medications. Raynaud phenomenon, migraine and valvular lesions were observed in 15% ( n = 10), 30% ( n = 20) and 18% ( n = 12) of the patients, respectively. Raynaud phenomenon and migraine, but not valvular lesions, were markedly more frequent in PAPS patients presenting with positive aPS/PT antibodies ( n = 10, 34.5% vs. n = 0, 0%; p = 0.0001). Conclusions In PAPS patients aPS/PT antibodies are related to the occurrence of both Raynaud phenomenon and migraine.

  15. Comparison of IgM Capture Enzyme- Linked Immunosorbent Assay (ELISA) using Inhouse method and commercially available MRL kit for serological confirmation of dengue infection

    International Nuclear Information System (INIS)

    Hapugoda, D.M.; De Silva R, Nilanthi; Abeywickreme, W.; Gunasena, Sunethra; Prithimala, L.D.; Jayawardene, S.L.G.J.; Kumari, Thamara

    2003-01-01

    Laboratory diagnosis of dengue infection is important for the management of the patients. In this study igM capture ELISA using an inhouse method and commercially available kit (MRL diagnostics,USA) was compared to detect diagnostic capability of Inhouse IgM ELISA for provision of diagnostic facilities to the public at an affordable cost. Eighty acute and convalescent serum samples were collected from serologically confirmed dengue patients. Serological confirmation of patients were performed by Haemagglutination Inhibition (HI) assay, gold standard assay for dengue on paired serum samples. All collected acute and convalescent sera were tested by IgM ELISA using the inhouse method and MRL kit. Antigen and conjugate for the inhouse IgM method were prepared in the laboratory. A cocktail of four dengue antigens containing 25 Antigen ELISA units of each type was prepared and used as the assay antigen. Conjugate was prepared using a serum sample with high dengue Anti flavi IgG antibody titre conjugated with Horseradish peroxidase. A prospective study of both IgM ELISA assays were performed using 113 acute sera collected from dengue suspected cases. Overall results showed that 46% and 52% acute sera collected from dengue confirmed patients were positive by inhouse ELISA assay and MRL kits respectively. In the prospective study done using acute sera collected from dengue suspected patients showed that 44% and 52% were positive by inhouse ELISA assay and MRL kits. There was no significant difference in positivity between these two assays. (P=0.18). Inhouse IgM ELISA can be used for provision of laboratory diagnosis of dengue virus infection more than 5 days. The assay is 10 times less costly than using MRL kits as assay antigen and conjugate can be prepared easily in the laboratory

  16. Anti-phospholipid antibodies in patients with Plasmodium falciparum malaria

    DEFF Research Database (Denmark)

    Jakobsen, P H; Morris-Jones, S D; Hviid, L

    1993-01-01

    Plasma levels of antibodies against phosphatidylinositol (PI), phosphatidylcholine (PC) and cardiolipin (CL) were measured by enzyme-linked immunosorbent assay (ELISA) in patients from malaria endemic area of Sudan and The Gambia. Some Sudanese adults produced IgM antibodies against all three types...... of phospholipids (PL) during an acute Plasmodium falciparum infection. The anti-PL antibody titre returned to preinfection levels in most of the donors 30 days after the disease episode. IgG titres against PI, PC and CL were low. In Gambian children with malaria, IgM antibody titres against PI and PC were...... significantly higher in those with severe malaria than in those with mild malaria. These results show that a proportion of malaria patients produce anti-PL antibodies during infection and that titres of these antibodies are associated with the severity of disease....

  17. Biochemical characterization and structure determination of a potent, selective antibody inhibitor of human MMP9.

    Science.gov (United States)

    Appleby, Todd C; Greenstein, Andrew E; Hung, Magdeleine; Liclican, Albert; Velasquez, Maile; Villaseñor, Armando G; Wang, Ruth; Wong, Melanie H; Liu, Xiaohong; Papalia, Giuseppe A; Schultz, Brian E; Sakowicz, Roman; Smith, Victoria; Kwon, Hyock Joo

    2017-04-21

    Matrix metalloproteinase 9 (MMP9) is a member of a large family of proteases that are secreted as inactive zymogens. It is a key regulator of the extracellular matrix, involved in the degradation of various extracellular matrix proteins. MMP9 plays a pathological role in a variety of inflammatory and oncology disorders and has long been considered an attractive therapeutic target. GS-5745, a potent, highly selective humanized monoclonal antibody inhibitor of MMP9, has shown promise in treating ulcerative colitis and gastric cancer. Here we describe the crystal structure of GS-5745·MMP9 complex and biochemical studies to elucidate the mechanism of inhibition of MMP9 by GS-5745. GS-5745 binds MMP9 distal to the active site, near the junction between the prodomain and catalytic domain, and inhibits MMP9 by two mechanisms. Binding to pro-MMP9 prevents MMP9 activation, whereas binding to active MMP9 allosterically inhibits activity. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. Transplantation of ABO A2 kidneys into O recipients: do IgM anti-A1 titers matter?

    Science.gov (United States)

    Tierney, Joshua; Shaffer, David

    2015-04-01

    The ABO blood subgroup A2 expresses lower levels of A antigen on the cell surface and is less immunogenic toward anti-A immunoglobulin present in blood type O or B recipients. Previous studies have shown successful kidney transplantation from A2 donors into O or B recipients with low pre-transplant anti-A titers. Previous studies suggest good results with recipient IgG titers A1 IgM titers on early outcomes following A2 to O or B kidney transplantation. We performed a single center, retrospective review of all A2 to O living donor kidney transplants. All recipients had pre-transplant anti-A IgG titers <1:8. IgM titers were measured in all recipients and were reported but not used to determine eligibility for transplant. From 2001 to 2013, we performed seven consecutive A2 to O living donor kidney transplants. Early allograft dysfunction, acute rejection or thrombotic microangiopathy, occurred in four patients and were associated with high IgM titers despite low IgG titers. Our data show a high incidence of early acute rejection or thrombotic microangiopathy in A2 to O kidney transplants with high recipient anti-A IgM titers despite low IgG titers. Steps to lower anti-IgM pre-transplant may reduce the risk of early allograft dysfunction in A2 to O or B kidney transplants. Attention should be paid to IgM titers in establishing individual center selection criteria for A2 to B kidney transplants under the new UNOS kidney allocation system. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  19. Increased susceptibility of tumor cells and chicken erythrocytes to lysis by antibody and complement after treatment with aminoethylisothiouronium bromide hydrobromide (AET)

    Energy Technology Data Exchange (ETDEWEB)

    Langone, J.J.; Borsos, T. (National Cancer Institute, Bethesda, MD (USA))

    1979-01-01

    After treatment with aminoethylisothiouronium bromide hydrobromide (AET), the ascites forms of the diethylnitrosamine-induced guinea pig hepatomas, line-1 and line-10, were susceptible or more susceptible to killing in vitro by certain combinations of tumor-specific or IgM anti-Forssman antibody and either human or guinea pig complement. Since AET could be toxic to either cell line, conditions of pH, concentration of AET, and duration of exposure of the cells to the reagent were determined that resulted in enhanced susceptibility without significantly affecting cell viability. Chicken erythrocytes (CE) also were tested and AET-treated cells found to be more susceptible to lysis by IgM anti-Forssman antibody and guinea pig complement. The enhancement apparently was not due to increased ability of AET-treated CE to fix antibody. In contrast to CE, the lytic susceptibility of sheep cells was not affected by AET treatment. In addition to AET, several drugs and enzymes that also can affect the susceptibility of the tumor cells to antibody and complement were tested and found to be ineffective against CE. Since AET reputedly acts directly on the cell surface, it seems reasonable to assume that the increased susceptibility of the tumor cells and CE to antibody and complement may result from a modification of the cell surface.

  20. Anticardiolipin and antinuclear antibodies in the adult healthy Omani individuals

    International Nuclear Information System (INIS)

    Al-Jabri, Ali A.; Al-Buloshi, Mohamed S.

    2004-01-01

    To investigate the prevalence and normal versus abnormal levels of anticardiolipin antibodies (aCL) in a healthy adult popultion of Omani's and whether a correlation exists in between aCL and antinuclear antibodies (ANA) in this Omani population. A total of 521 healthy Omani individuals (333 males and 188) females aged between 17-54 years were investigated for the presence and quntities of aClL (immunoglubolin G (IgG)) and IgM isotypesusing a conventional enzyme linked immunosobrent assay. ANA were were detacted in this group, using standard indirect immunofloresence techniques. This study was conducted during the period 2002 through 2003 at the Immunology Laboratories, College of Medicine and Health Sciences, Sultan Qaboos University, Al Khod, Muscat, Sultanate of Oman. The prevalence of of aCL in healthy Omani population was estimated to be 2.5% for IgG 3.1% for IgM. The cut off points for IgG and IgM were determined for the whole population as 22.5 IgG phospholipid (GPL) units and 15.7 IgM phospholipid (MPL) units, using the mean plus 5 standard deviations. Using these cut off values, aCL were not detected in the the majority of individuals (97%) and in the remainig 3% the levels were not very high. There was no significant difference between the levels of aCL in either the male or female groups and no significant correlation for the presence of aCL with the age in this studied population. ANA were detected in 76/521 (14.6%)of the population studied, with some individuals studied, with some individuals (0.8%) showing titers of 1:640, but there was no association with aCL. Although ANA is present in this healthy Omani population at high frequency and some individuals at high levels of aCL do not occur and their presence may be an indicator of autoimmune mediated pathalogy. (author)

  1. Presence of anti-Toxoplasma antibodies in humans and their cats in the urban zone of Guadalajara

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    Galván Ramírez María de la Luz

    1999-01-01

    Full Text Available Cats are the definitive hosts of Toxoplasma gondii. Infected cats excrete oocysts in their feces, infecting humans and other animals. The objective of the present study was to determine the presence of anti-Toxoplasma antibodies in cat owners and their pets, and determine if there was a relationship between Toxoplasma infection and humans who live with infected cats. IgG anti-Toxoplasma antibodies in sera of 59 cat owners were determined by enzyme-linked immunosorbent assay (ELISA, in 24 sera from their cats, IgG, IgM, and IgA antibodies were found using Burney's ELISA. Thirty-eight (64% of 59 cat owners were positive to IgG anti-Toxoplasma. Seropositivity for cats was 70.8% IgG, 8.3% IgM, and 62.5% IgA. Cohabitation with cats infected by T. gondii, feeding with leftovers or raw viscera, and lack of control over how their feces were handled are risk factors conducive for humans to become infected by T. gondii.

  2. Western blot can distinguish natural and acquired antibodies to Mycoplasma agassizii in the desert tortoise (Gopherus agassizii).

    Science.gov (United States)

    Hunter, Kenneth W; Dupré, Sally A; Sharp, Tiffanny; Sandmeier, Franziska C; Tracy, C Richard

    2008-12-01

    Mycoplasma agassizi has been identified as a cause of upper respiratory tract disease (URTD) in the threatened Mojave population of the desert tortoise (Gopherus agassizii), and anti-M. agassizii antibodies have been found by ELISA in as many as 15% of these animals across their geographic range. Here we report that a cohort of 16 egg-reared desert tortoises never exposed to M. agassizii had ELISA antibody titers to this organism that overlapped with titers obtained from some M. agassizii-infected tortoises. These natural antibodies were predominantly of the IgM class. Western blots of plasma from these non-infected tortoises produced a characteristic banding pattern against M. agassizii antigens. A group of 38 wild-caught desert tortoises was tested by ELISA, and although some of these tortoises had antibody titers significantly higher than the non-infected tortoises, there was considerable overlap at the lower titer levels. However, Western blot analysis revealed distinct banding patterns that could readily distinguish between the non-infected tortoises and tortoises with acquired antibodies, regardless of ELISA antibody titers. We conclude that desert tortoises have natural antibodies to M. agassizii that can compromise the determination of infection status by ELISA. However, the Western blot technique can distinguish between natural and acquired antibody patterns and can be used to confirm the diagnosis of M. agassizii infections in the desert tortoise.

  3. Immunoglobulin M and G antibody responses to Plasmodium falciparum glutamate-rich protein

    DEFF Research Database (Denmark)

    Dziegiel, Morten Hanefeld; Rowe, P; Bennett, S

    1993-01-01

    The aims of the present study were to describe the age-related immunoglobulin M (IgM) and IgG response to part of a 220-kDa glutamate-rich protein (GLURP) from Plasmodium falciparum and to determine possible correlations of possession of these antibodies with malaria morbidity. IgM and IgG levels...... were measured with a recombinant fusion protein consisting of the carboxy-terminal 783 amino acids of the GLURP. Samples for the study were obtained during a longitudinal malaria morbidity survey performed in The Gambia; cross-sectional surveys were performed at the beginning of the transmission season...... in May and in October. Seropositivity rates increased with age to a maximum of 77% for IgM and 95% for IgG in adults. High prevalences of seropositivity were associated with certain human leukocyte antigen class II alleles (DRw8, DR9, DR7, DR4, DQw7, and DQw2) or haplotypes. The relationship between anti...

  4. Antibody and cytokine serum levels in patients subjected to anti-rabies prophylaxis with serum-vaccination

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    J. A. Ayres

    2006-01-01

    Full Text Available Rabies is considered a fatal disease once clinical symptoms have developed. The aim of this study was to evaluate epidemiological aspects and immune response in patients attacked by domestic and wild animals and subjected to post-exposure rabies treatment with equine serum and associated vaccine. Thirty-three patients were evaluated; they were between 13 and 65 years old, 75.8% were male and 24.2% female, and from the Botucatu neighborhood. Twenty healthy control individuals with the same age range were also studied. Specific antibodies to equine immunoglobulins and IFN-gamma, IL-2, IL-4, and IL-10 production were evaluated by ELISA. IgM, IgE, IgG and subclasses, and rabies virus antibodies serum levels were determined by nephelometry and seroneutralization methods, respectively. No anaphylactic or serum sickness allergic reactions were observed in patients after treatment. Anti-equine IgG levels were significantly higher than those of IgM after 14 and 28 days of treatment. Protective antibodies to rabies virus > 0.5 UI/ml were detected in 84.6% and 75% of patients at days 14 and 28, respectively. IFN-gamma, IL-2 and IL-10 levels in patients before and 48h after treatment were significantly higher than in controls suggesting that both Th1 and Th2 cells were activated in the patients. Serum IgM levels were higher at day 14, and IgG2 and IgE levels were higher at day 28 of treatment. These results suggest that post-exposure rabies treatment in humans induces significant alterations in patient immune response characterized by increased levels of cytokines, serum levels of specific rabies virus antibodies, and the equine serum components employed in the treatment.

  5. Synthetic rabbit-human antibody conjugate as a control in immunoassays for immunoglobulin M specific to hepatitis E virus

    Directory of Open Access Journals (Sweden)

    Zhang Rui

    2010-05-01

    Full Text Available Abstract Background In assays for anti-hepatitis E virus (HEV immunoglobulin M (IgM, large volumes of the patient's sera cannot be easily obtained for use as a positive control. In this study, we investigated an alternative chemical method in which rabbit anti-HEV IgG was conjugated with human IgM and was used as a positive control in the anti-HEV IgM assay. Rabbit anti-HEV IgG was isolated from immune sera by chromatography on protein A-Sepharose and was conjugated with human IgM by using 1-ethyl-3-(3-dimethylaminopropylcarbodiimide (EDC as a crosslinker. Results The specific anti-HEV IgG antibody titer was 100,000 times that of the negative control, i.e., prebleed rabbit serum. The results of anti-HEV IgM enzyme-linked immunosobent assay showed that the antibody conjugate was similar to anti-HEV IgM antibodies produced in humans. The results of a stability experiment showed that the antibody conjugate was stable for use in external quality assessment or internal quality control trials. Conclusions We concluded that the chemically conjugated rabbit-human antibody could be used instead of the traditional serum control as a positive control in the anti-HEV IgM assay.

  6. Detection of anti neutrophil antibodies by radio-iodinated protein A binding test

    International Nuclear Information System (INIS)

    Cartron, J.; Muller, J.Y.; Tchernia, G.; Paule, B.; Varet, B.

    1983-01-01

    The granulocyte associated IgG in normal and neutropenic subjects has been determined by a direct quantitative assay using radiolabeled staphylococcal protein A. This assay allows to postulate an immunological mechanism to explain the neutropenia in 19 cases of neutropenias associated with malfunctions of the immune system and in 4 cases of idiopathic neutropenias. Discussed in this report is the possible interaction of immune complexes bound in vivo to the granulocytes. By an immunofluorescence test, it has been possible to detect IgG or IgM antibodies in only 52% of patients with a positive direct assay. The determination of granulocyte-associated IgG is therefore a better indicator for defining an auto-immune neutropenia than the detection of free antibodies in the sera [fr

  7. Glomerulonephritis in schistosomiasis with mesangial IgM deposits

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    Ageu Godoy Magalhães Filho

    1981-06-01

    Full Text Available Twenty one cases of hepatoesplenic schistosomiasis patients without clinical and laboratory evidence of renal disease, were studied by surgical biopsies using light microscopy and immunofluorescence. The cases were classified histologically as: normal pattern (6 cases; minimal changes (6 cases; and mesangial proliferative glomerulonephritis (9 cases. By the immunofluorescence microscopy using anti IgM, IgG, IgA and C3, the predominant finding in all biopsies, except the normal cases, was granular deposits of IgM in the mesangium along with C3. On the other hand, IgG was present in all cases including normal biopsies along the capillary walls. However IgG was also present in the mesangium only in cases with glomerular lesions. This finding may well be similar to that recently described as IgM mesangial nephropathy. According to our cases a mesangial proliferative glomerulonephritis, characterized by segmental cell proliferation and deposition of IgM in the mesangium, is probably the entity found in the early stages of mansonic schistosomiasis.Com o objetivo de observar as lesões renais iniciais na esquistossomose mansoni, foram selecionados 21 casos da forma hepato-esplênica, com idade média de 12 anos e que não apresentavam sinais clínicos ou laboratoriais de alterações renais. As biópsias foram realizadas durante esplenectomia e em seguida examinadas à microscopia ótica e à microscopia de fluorescência, sendo empregados conjugados anti IgG, IgM, IgA e C3, isoladamente. No exame microscópico, os 21 casos foram classificados em: 6 casos normais; 6 casos com lesões glomerulares consideradas mínimas e 9 casos com lesões mesangiais discretas ou moderadas. À imunofluorescência constatou-se presença de depósitos de IgM e de C3 predominantemente de localização mesangial, tanto nos casos de lesões mínimas, como nos casos de proliferação mesangial mais acentuada (Tabelas II e III. Constatou-se aind a depósitos de IgG em todos

  8. Evaluation of the clinical relevance of anti-annexin-A5 antibodies in Chinese patients with antiphospholipid syndrome.

    Science.gov (United States)

    Zhang, Shulan; Wu, Ziyan; Li, Jing; Wen, Xiaoting; Li, Liubing; Zhang, Wen; Zhao, Jiuliang; Zhang, Fengchun; Li, Yongzhe

    2017-02-01

    A hallmark feature of antiphospholipid syndrome (APS) is the presence of antiphospholipid antibodies (aPLs). Few studies have addressed the clinical relevance of anti-annexin A5 antibodies (aANXA5) in Chinese patients with APS. In this study, we evaluated the clinical performance of aANXA5 in the diagnosis of APS. Sera from 313 subjects were tested, including 170 samples from patients with APS, 104 samples from patients with non-APS diseases as disease controls (DC), and 39 healthy controls (HC). Serum IgG and IgM aANXA5 were determined by ELISA. Overall, the levels of both IgG and IgM aANXA5 were significantly increased in patients with primary APS (PAPS) and APS associated to other diseases (APSAOD) compared with DC and HC. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for IgG and IgM aANXA5 in the diagnosis of APS were 33.5 and 15.3, 99.0 and 99.0, 98.3 and 96.3, and 47.7 and 41.7%, respectively. Significant associations between IgG aANXA5 and arterial thrombotic events (OR, 2.60; 95% CI, 1.44-4.71) and between IgG aANXA5 and venous thrombotic events (OR, 2.80; 95% CI, 1.55-5.06) were identified. No correlations were identified between IgG or IgM aANXA5 and obstetric complications. Our data suggest that aANXA5 could serve as a diagnosis biomarker for patients with APS. More importantly, our data highlighted a potential role of IgG aANXA5 in identifying APS patients with high risk of thrombosis.

  9. Species specificity of a monoclonal antibody produced to Naegleria fowleri and partial characterization of its antigenic determinant.

    Science.gov (United States)

    Réveiller, F L; Marciano-Cabral, F; Pernin, P; Cabanes, P A; Legastelois, S

    2000-08-01

    Monoclonal antibody (Mab) 5D12 against Naegleria fowleri was analyzed for species specificity. Mab 5D12 reacted with a ubiquitous epitope present on the membrane of N. fowleri but not with soluble antigens. The Mab did not react with N. lovaniensis, N. gruberi, N. australiensis, or Acanthamoeba castellanii. The decreased reactivity of Mab 5D12 with N. fowleri observed after periodate oxidation, after digestion of carbohydrate moieties by three glycosidases, or after treatment of amebas with tunicamycin strongly suggests that the antigenic determinant has a polysaccharide component. Inhibition of the reactivity of Mab 5D12 by soluble saccharides supports the idea that N-acetyl or amino groups may play an important role in the recognition of the carbohydrate component of the epitope by the Mab. The specificity of Mab 5D12 makes this an ideal reagent for the identification of N. fowleri in environmental samples or in clinical specimens.

  10. Generation of polyclonal antibody with high avidity to rosuvastatin and its use in development of highly sensitive ELISA for determination of rosuvastatin in plasma

    Directory of Open Access Journals (Sweden)

    Al-Malaq Hamoud A

    2011-07-01

    Full Text Available Abstract In this study, a polyclonal antibody with high avidity and specificity to the potent hypocholesterolaemic agent rosuvastatin (ROS has been prepared and used in the development of highly sensitive enzyme-linked immunosorbent assay (ELISA for determination of ROS in plasma. ROS was coupled to keyhole limpt hemocyanin (KLH and bovine serum albumin (BSA using carbodiimide reagent. ROS-KLH conjugate was used for immunization of female 8-weeks old New Zealand white rabbits. The immune response of the rabbits was monitored by direct ELISA using ROS-BSA immobilized onto microwell plates as a solid phase. The rabbit that showed the highest antibody titer and avidity to ROS was scarified and its sera were collected. The IgG fraction was isolated and purified by avidity chromatography on protein A column. The purified antibody showed high avidity to ROS; IC50 = 0.4 ng/ml. The specificity of the antibody for ROS was evaluated by indirect ELISA using various competitors from the ROS-structural analogues and the therapeutic agents used with ROS in a combination therapy. The proposed ELISA involved a competitive binding reaction between ROS, in plasma sample, and the immobilized ROS-BSA for the binding sites on a limited amount of the anti-ROS antibody. The bound anti-ROS antibody was quantified with horseradish peroxidase-labeled second anti-rabbit IgG antibody (HRP-IgG and 3,3',5,5'-tetramethylbenzidine (TMB as a substrate for the peroxidase enzyme. The concentration of ROS in the sample was quantified by its ability to inhibit the binding of the anti-ROS antibody to the immobilized ROS-BSA and subsequently the color intensity in the assay wells. The assay enabled the determination of ROS in plasma at concentrations as low as 40 pg/ml.

  11. Missing Links in Antibody Assembly Control

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    Tiziana Anelli

    2013-01-01

    Full Text Available Fidelity of the humoral immune response requires that quiescent B lymphocytes display membrane bound immunoglobulin M (IgM on B lymphocytes surface as part of the B cell receptor, whose function is to recognize an antigen. At the same time B lymphocytes should not secrete IgM until recognition of the antigen has occurred. The heavy chains of the secretory IgM have a C-terminal tail with a cysteine instead of a membrane anchor, which serves to covalently link the IgM subunits by disulfide bonds to form “pentamers” or “hexamers.” By virtue of the same cysteine, unassembled secretory IgM subunits are recognized and retained (via mixed disulfide bonds by members of the protein disulfide isomerase family, in particular ERp44. This so-called “thiol-mediated retention” bars assembly intermediates from prematurely leaving the cell and thereby exerts quality control on the humoral immune response. In this essay we discuss recent findings on how ERp44 governs such assembly control in a pH-dependent manner, shuttling between the cisGolgi and endoplasmic reticulum, and finally on how pERp1/MZB1, possibly as a co-chaperone of GRP94, may help to overrule the thiol-mediated retention in the activated B cell to give way to antibody secretion.

  12. Primary alveolar echinococcosis: course of larval development and antibody responses in intermediate host rodents with different genetic backgrounds after oral infection with eggs of Echinococcus multilocularis.

    Science.gov (United States)

    Matsumoto, Jun; Kouguchi, Hirokazu; Oku, Yuzaburo; Yagi, Kinpei

    2010-09-01

    We investigated parasite establishment, subsequent larval development and antibody responses in gerbils, cotton rats and 4 inbred mouse strains until 16 weeks post inoculation (p.i.) with 200 eggs of Echinococcus multilocularis. The rate of parasite establishment in the liver determined at 4 weeks p.i. was highest in DBA/2, followed by AKR/N, C57BL/10 and C57BL/6 mice, whereas gerbils harboured few parasite foci. The accurate number of liver lesions in cotton rats could not be determined due to rapid growth and advanced multivesiculation of the parasite observed at 2 weeks p.i. The course of larval development was most advanced in DBA/2 mice with mature protoscolex formation at 16 weeks p.i., followed by AKR/N harbouring metacestodes with sparsely distributed immature protoscoleces. On the other hand, C57BL/6 and C57BL/10 mice had infertile metacestodes without any protoscolex formation. The parasite growth in mice was totally slower than those in gerbils and cotton rats. Specific IgG and IgM responses against 3 types of native crude antigens of larval E. multilocularis were evaluated using somatic extracts of and vesicle fluid of metacestode, and somatic extracts from purified protoscoleces. The 4 mouse strains demonstrated basically similar kinetics with apparent IgG and IgM increases at 9 weeks p.i. and thereafter, except C57BL/10, exhibited higher levels of IgM against crude antigens at some time point of infection. On the other hand, a follow-up determination of specific IgG and IgM levels against recombinant antigens from larval E. multilocularis revealed that each mouse strain showed different antibody-level kinetics. The findings in the present study demonstrate that the course of host-parasite interactions in primary alveolar echinococcosis, caused by larval E. multilocularis, clearly varies among intermediate host rodents with different genetic backgrounds.

  13. X-linked Hyper IgM Syndrome Presenting as Pulmonary Alveolar Proteinosis.

    Science.gov (United States)

    Gallagher, Joel; Adams, Juan; Hintermeyer, Mary; Torgerson, Troy R; Lopez-Guisa, Jesus; Ochs, Hans D; Szabo, Sara; Salib, Mina; Verbsky, James; Routes, John

    2016-08-01

    X-linked hyper IgM syndrome (XHIGM) is a combined immunodeficiency caused by mutations in the CD40 ligand (CD40L) gene that typically results in decreased or absent CD40L expression on activated T cells, leading to defective class switching and somatic hypermutation. We describe an infant who presented with respiratory failure due to pulmonary alveolar proteinosis (PAP) with a novel damaging missense mutation in the CD40L gene. Whole exome sequencing (WES) was used to identify a mutation in the CD40L gene. CD40L expression and function were determined by flow cytometry. A 5-month-old previously-healthy male presented with respiratory failure and diffuse pulmonary ground glass opacities on CT scan of the chest. Laboratory evaluation revealed an undetectable IgG, normal IgA, and elevated IgM. A bronchoalveolar lavage demonstrated pulmonary alveolar proteinosis. WES demonstrated a c.608G > C mutation in the CD40L gene resulting in p.R203T. Flow cytometry demonstrated normal CD40L expression on activated T cells but absent binding of CD40-Ig to CD40L on activated patient T cells. The clinical manifestations of XHIGM in our patient had several unique features, including the presentation with PAP, normal serum IgA, and expression of non-functional CD40L on activated T cells. To our knowledge, this is the first published case of PAP in a patient with XHIGM.

  14. Production and Purification of Monoclonal Antibody Against Tumor Marker of TPA

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    Seyyed Amir Abbas Ghodrat

    2016-05-01

    Full Text Available Considering the invasive nature of cancer cells, one of the most important and best indicator of them is the markers inside them. One of the most important markers that observed in some types of cancer cells in various parts of the body is the Cytokeratin. Tissue plasminogen activator antigen (TPA is a Cytokeratin composed of molecules with various molecular weights. The level of TPA serum as associated with cellular growth level and tumorization of cells. In this research, the hybrid of spleen cells in BALB/c female mouse with myeloma cells was conducted with a ratio of 10:1. The resulting monoclonal antibodies were confirmed by SDS-PAGE and western blot. Protein G chromatography was utilized to purify monoclonal antibodies. The results for determining isotypes showed IgM and IgG classes. The titer of the antibody obtained from various clones was capable of identifying Cytokeratin antigen with a dilution of 1/10000. The resulting antibodies were finally confirmed by western blot and all the 5 resulting monoclonal antibodies were capable of identifying a 48 kDa protein. The results indicate that with the help of TPA marker and the monoclonal antibodies produced against them, this marker can be recognized quickly with great accuracy in suspicious cases of cancer. Thus, appropriate measures will be taken to prevent and fight off its probable side effects. This factor can be further used to build a diagonal kit with high sensitivity.

  15. A public antibody lineage that potently inhibits malaria infection through dual binding to the circumsporozoite protein.

    Science.gov (United States)

    Tan, Joshua; Sack, Brandon K; Oyen, David; Zenklusen, Isabelle; Piccoli, Luca; Barbieri, Sonia; Foglierini, Mathilde; Fregni, Chiara Silacci; Marcandalli, Jessica; Jongo, Said; Abdulla, Salim; Perez, Laurent; Corradin, Giampietro; Varani, Luca; Sallusto, Federica; Sim, Betty Kim Lee; Hoffman, Stephen L; Kappe, Stefan H I; Daubenberger, Claudia; Wilson, Ian A; Lanzavecchia, Antonio

    2018-03-19

    Immunization with attenuated Plasmodium falciparum sporozoites (PfSPZs) has been shown to be protective against malaria, but the features of the antibody response induced by this treatment remain unclear. To investigate this response in detail, we isolated IgM and IgG monoclonal antibodies from Tanzanian volunteers who were immunized with repeated injection of Sanaria PfSPZ Vaccine and who were found to be protected from controlled human malaria infection with infectious homologous PfSPZs. All isolated IgG monoclonal antibodies bound to P. falciparum circumsporozoite protein (PfCSP) and recognized distinct epitopes in its N terminus, NANP-repeat region, and C terminus. Strikingly, the most effective antibodies, as determined in a humanized mouse model, bound not only to the repeat region, but also to a minimal peptide at the PfCSP N-terminal junction that is not in the RTS,S vaccine. These dual-specific antibodies were isolated from different donors and were encoded by VH3-30 or VH3-33 alleles that encode tryptophan or arginine at position 52. Using structural and mutational data, we describe the elements required for germline recognition and affinity maturation. Our study provides potent neutralizing antibodies and relevant information for lineage-targeted vaccine design and immunization strategies.

  16. مناسبترین زمان جمع آوری نمونه بعد از شروع یکباره بیماری جهت تشخیص پادتن IgM سرخک

    Directory of Open Access Journals (Sweden)

    طلعت مختاری آزاد

    2005-02-01

    Full Text Available Antibody detection is the most versatile and commonly used method for measles diagnosis. Detection of specific IgM antibodies in a single serum specimen collected within the appropriate time after rash onset can provide a good presumptive diagnosis of current or recent measles infection and is the test of choice for rapid diagnosis of measles cases. So, optimal timing for collection of a single serum specimen to diagnose measles by IgM capture Enzyme Immunoassay (EIA was evaluated. 399-paired sera were tested for measles IgM antibody. 149-paired sera were measles IgM negative. Two hundred fifty paired sera had at least one IgM positive. 223-paired sera were positive in both first and second samples. 19-paired sample were negative in first and positive in second sample and 8-paired sera were positive in the first and negative in the second samples. 85% and 100% of first specimens within 7 and 7-21 and 94% of all second samples were IgM positive at 28th days after rash onset, respectively. Analysis of data indicates that a single serum specimen collected between 7 to 27 days after rash onset can be used to diagnose most cases of measles with an IgM capture EIA.

  17. Using molecular principal axes for structural comparison: determining the tertiary changes of a FAB antibody domain induced by antigenic binding

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    Silverman B David

    2007-11-01

    Full Text Available Abstract Background Comparison of different protein x-ray structures has previously been made in a number of different ways; for example, by visual examination, by differences in the locations of secondary structures, by explicit superposition of structural elements, e.g. α-carbon atom locations, or by procedures that utilize a common symmetry element or geometrical feature of the structures to be compared. Results A new approach is applied to determine the structural changes that an antibody protein domain experiences upon its interaction with an antigenic target. These changes are determined with the use of two different, however comparable, sets of principal axes that are obtained by diagonalizing the second-order tensors that yield the moments-of-geometry as well as an ellipsoidal characterization of domain shape, prior to and after interaction. Determination of these sets of axes for structural comparison requires no internal symmetry features of the domains, depending solely upon their representation in three-dimensional space. This representation may involve atomic, Cα, or residue centroid coordinates. The present analysis utilizes residue centroids. When the structural changes are minimal, the principal axes of the domains, prior to and after interaction, are essentially comparable and consequently may be used for structural comparison. When the differences of the axes cannot be neglected, but are nevertheless slight, a smaller relatively invariant substructure of the domains may be utilized for comparison. The procedure yields two distance metrics for structural comparison. First, the displacements of the residue centroids due to antigenic binding, referenced to the ellipsoidal principal axes, are noted. Second, changes in the ellipsoidal distances with respect to the non-interacting structure provide a direct measure of the spatial displacements of the residue centroids, towards either the interior or exterior of the domain

  18. Antibodies, directed towards Campylobacter jejuni antigens, in sera from poultry abattoir workers

    Science.gov (United States)

    Cawthraw, S A; Lind, L; Kaijser, B; Newell, D G

    2000-01-01

    Occupational exposure of susceptible humans to Campylobacter jejuni appears to result in resistance to disease. This is believed to be due to acquired protective immunity. To support this hypothesis the levels of C. jejuni-specific IgG and IgM antibodies were determined in sera from poultry abattoir workers. Such individuals are persistently exposed to C. jejuni, but apparently rarely acquire campylobacteriosis. Sera from 43 short-term workers (employed ≤1 month), 78 long-term workers and 40 blood donors were investigated by ELISA. In 51 individuals a second serum sample, taken at least 1 month after the first, was also investigated. Eight workers had C. jejuni-positive faecal cultures and only one, a short-term worker, had symptoms of campylobacteriosis. There were significantly higher levels of specific IgG antibodies in long-term workers than in either of the other groups. There was no significant difference detectable in specific IgM antibody levels between any of the groups. The results provide supporting evidence that long-term exposure to C. jejuni induces circulating antibodies which reflect apparent reduced susceptibility to disease. Western blotting showed flagellin and polypeptides of 45, 40, 32 and 30 kD bound antibodies significantly more frequently by sera from long-term workers than short-term workers and blood donors. The most commonly detected antigens were the 40-kD (80%) and flagellin (55%). The results indicate that specific serum IgG responses induced by endemic exposure to C. jejuni might be directed towards a small number of protein antigens with apparently conserved epitopes. PMID:11012618

  19. Ri antibodies in patients with breast, ovarian or small cell lung cancer determined by a sensitive immunoprecipitation technique.

    Science.gov (United States)

    Knudsen, Anette; Monstad, Sissel E; Dørum, Anne; Lønning, Per E; Salvesen, Helga B; Drivsholm, Lars; Aarseth, Jan H; Vedeler, Christian A

    2006-10-01

    The presence of circulating antineuronal antibodies has been associated with paraneoplastic neurological syndromes (PNS). Ri antibodies are often associated with lung or breast cancer, but the prevalence of such antibodies in large cancer materials is largely unknown. We used a highly sensitive immunoprecipitation assay to study the level of Ri antibodies in blood samples from 200 patients with small cell lung cancer (SCLC), 253 patients with breast cancer and 557 patients with ovarian cancer. Two hundred blood donors and six Ri positive PNS patients served as controls. The recombinant antigen used in the immunoprecipitation assay was radiolabeled by a coupled in vitro transcription and translation (ITT) technique, enabling low levels of antibodies to be detected. None of the blood donors contained Ri antibodies, whereas all of the sera from the PNS patients were positive. Ri antibodies were present in 4.5% of the patients with SCLC, 0.8% of the patients with breast cancer and in 0.2% of the patients with ovarian cancer. Retesting of the Ri positive samples with immunofluorescense and immune blot showed that the immunoprecipitation technique was more sensitive than the other immune assays. Ri antibodies were not associated with PNS in the patients with breast or ovarian cancer. Neurological data were not available for the SCLC patients, but in these, Ri antibodies were not associated with survival.

  20. Frequency of Anti-Toxoplasma IgM and IgG Antibodies in Leukemic Patients

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    M.J. Gharavi

    2015-11-01

    Full Text Available Aim: the efficacy and safety of combined therapy with anticholinergics and alpha blockers in comparison with alpha blockers monotherapy among patients with BPH. Methods: We conducted a single blinded RCT and 80 patients with BPH who referred to Ali-ebn-abitaleb, Zahedan were included. One group received 0.4 mg/d Tamsulosin in addition to Tolterodine and the other group received the same doses of Tamsulosin in addition to placebo. Both groups were evaluated before and 6 months after treatment with special attention to clinical symptoms, life quality improvement and treatment satisfaction. SPSS was applied for data analysis. Results: There is no significant difference between these two groups of patients’ demographics and basic indices such as PVR, PSA and the volume of prostate. IPSS was significantly improved among patients who received Toltrodine (p=0.0008 whereas both groups showed same score before treatment. Conclusion: It seems that combined therapy with anticholinergics and alpha blockers is a safe and efficient regimen among patients with BPH, either as primary or alternative treatment after the failure of conventional treatments.

  1. Purification of Monoclonal Antibodies Using a Fiber Based Cation-Exchange Stationary Phase: Parameter Determination and Modeling

    Directory of Open Access Journals (Sweden)

    Jan Schwellenbach

    2016-10-01

    Full Text Available Monoclonal antibodies (mAb currently dominate the market for protein therapeutics. Because chromatography unit operations are critical for the purification of therapeutic proteins, the process integration of novel chromatographic stationary phases, driven by the demand for more economic process schemes, is a field of ongoing research. Within this study it was demonstrated that the description and prediction of mAb purification on a novel fiber based cation-exchange stationary phase can be achieved using a physico-chemical model. All relevant mass-transport phenomena during a bind and elute chromatographic cycle, namely convection, axial dispersion, boundary layer mass-transfer, and the salt dependent binding behavior in the fiber bed were described. This work highlights the combination of model adaption, simulation, and experimental parameter determination through separate measurements, correlations, or geometric considerations, independent from the chromatographic cycle. The salt dependent binding behavior of a purified mAb was determined by the measurement of adsorption isotherms using batch adsorption experiments. Utilizing a combination of size exclusion and protein A chromatography as analytic techniques, this approach can be extended to a cell culture broth, describing the salt dependent binding behavior of multiple components. Model testing and validation was performed with experimental bind and elute cycles using purified mAb as well as a clarified cell culture broth. A comparison between model calculations and experimental data showed a good agreement. The influence of the model parameters is discussed in detail.

  2. Antibody responses to Rhoptry-Associated Protein-1 (RAP-1) of Plasmodium falciparum parasites in humans from areas of different malaria endemicity

    DEFF Research Database (Denmark)

    Jakobsen, P H; Kurtzhals, J A; Riley, E M

    1997-01-01

    Plasma IgM and IgG antibody reactivities against the recombinant Plasmodium falciparum protein, Rhoptry Associated Protein-1 (rRAP-1) were measured by ELISA in individuals from Sudan, Indonesia, Kenya and The Gambia living in areas of different malaria endemicity. IgG and IgM reactivities to rRAP...

  3. Application of solid-phase radioimmunoassay in determining antibodies to Aujeszky's disease virus in blood serum of vaccinated pigs

    Energy Technology Data Exchange (ETDEWEB)

    Rodak, L.; Smid, B.; Valicek, L. (Vyzkumny Ustav Veterinarniho Lekarstvi, Brno-Medlanky (Czechoslovakia))

    1983-11-01

    In the blood sera of pigs vaccinated with inactivated vaccines manufactured by three different manufacturers the RIA method was used to determine the specific antibodies to the virus of Aujeszky's disease. In certain groups of vaccinated pigs the results of the RIA examination are unfavourably affected by the bond of antibodies to the cellular antigenous determinants. This proves that following vaccination antibodies are formed not only against the viral antigen but also against the antigens of cells on which the vaccination virus is propagated. These shortcomings are eliminated by the use of suitable cellular cultures for the preparation of viral and control antigens. Antigens are applicable for RIA and for ELISA examinations of blood sera of infected and vaccinated pigs. The advantages are described of the RIA and ELISA methods as compared with the virus neutralization test.

  4. Antibodies directed against monomorphic and evolutionary conserved self epitopes may be generated in 'knock-out' mice. Development of monoclonal antibodies directed against monomorphic MHC class I determinants

    DEFF Research Database (Denmark)

    Claesson, M H; Endel, B; Ulrik, J

    1994-01-01

    binding antibodies. Three stable anti-MHC class I (MHC-I) antibody secreting hybridoma clones were established and subcloned. All three MoAbs precipitated radiolabelled H-2 molecules as analysed by SDS PAGE, and all three MoAbs stained H-2b, H-2d, as well as H-2k cells by FACS analysis. The MoAbs stained...... to two beta 2m loss mutant cell lines, C4.4-25- and R1E, suggesting that some MHC-I heavy chain is exported to the cell surface even in the absence of endogenous beta 2m. Staining of murine cell lines kept under serum-free culture conditions was strongly influenced by the addition of bovine or human......-S cells were cultured at 37 degrees C. In total, these observations suggest that the MoAbs recognize conformational, presumably beta 2m and peptide dependent, self epitopes on MHC-class I. One of the three MoAbs stained rat blood mononuclear blood cells (BMC), all three MoAbs stained hamster BMC, whereas...

  5. Evaluation of Antigen-Specific IgM and IgG Production during an In Vitro Peripheral Blood Mononuclear Cell Culture Assay

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    Yoshiko Matsuda

    2017-07-01

    Full Text Available The recent attention given to diseases associated with memory B-cell (mBC-produced antibodies (Abs suggests the need for a similar in vitro assay to evaluate the functions of mBCs. Here, we cultured peripheral blood mononuclear cells (PBMCs with the intent to collect mBC-derived Abs in vitro and maintain their cell–cell contact-dependent interactions with helper T-cells. PBMCs were cultured with interleukin (IL-21, CpG-oligodeoxynucleotides (ODN, phorbol myristate acetate (PMA, and phytohemagglutinin/leucoagglutinin (PHA-L in 24-well flat-bottom plates (5 × 105 cells/well. A culture supernatant analysis of PBMCs from healthy donors (n = 10 indicated that antigen-specific IgM Ab levels in a PBMC culture supernatant might be better able to demonstrate the antigen sensitization status in a smaller peripheral blood sample, compared to IgG because Epstein–Barr virus-specific IgM mBCs circulate peripherally at a significantly higher frequency once antiviral humoral immunity has stabilized. Thus, our in vitro assay demonstrated the potential significance of antigen-specific IgM Ab production in the culture supernatants. Furthermore, an analysis of cultured PBMCs from allograft kidney recipients (n = 16 sensitized with de novo donor-specific human leukocyte antigen (HLA-specific Abs (DSAs showed that IgM-type HLA-specific Abs were detected mainly from the culture supernatants from PBMCs of patients with stable graft function, whereas IgG isotype HLA Abs were detectable only from patients with biopsy-proven antibody-mediated rejection. In other words, these IgG isotype Abs also represented an activated humoral immune response in vivo. Additionally, IgM- and IgG-expressing mBCs from healthy donors (n = 5 were cultured with IL-21, CpG-ODN, and a supernatant produced by stimulating CD19+ B-cell-depleted PBMCs with PHA-L and PMA in 24-well flat-bottom plates (1 × 105 cells/well, and the resulting in vitro analysis provided some

  6. Antiphospholipid Antibody Titers and Clinical Outcomes in Patients with Recurrent Miscarriage and Antiphospholipid Antibody Syndrome: A Prospective Study

    Science.gov (United States)

    Song, Yu; Wang, Hai-Yan; Qiao, Jie; Liu, Ping; Chi, Hong-Bin

    2017-01-01

    Background: The management of patients with recurrent miscarriage (RM) and antiphospholipid antibody syndrome (APS) includes prolonged treatment with heparin and aspirin, starting from the confirmation of pregnancy and continuing until 6 weeks after birth. This study was conducted to determine the relationship between changes in antiphospholipid antibody titers and clinical outcomes. The effect of a shortened treatment regimen was also evaluated. Methods: A prospective study of 123 patients with RM and APS between March 2012 and May 2014 was conducted. Patients were pretreated with a low dose of prednisone plus aspirin before pregnancy, and heparin was added after conception. The levels of antiphospholipid antibodies and pregnancy outcomes were evaluated. Results: All patients were positive for anti-β2-glycoprotein 1 (anti-β2-GP1) IgM. After prepregnancy treatment with low-dose prednisone plus aspirin, 99 of 123 patients became pregnant, and 87 of those pregnancies resulted in successful live births, while 12 resulted in miscarriage, showing a success rate of 87.9%. In the live birth group, levels of anti-β2-GP1 were 56.8 ± 49.0 RU/ml before the pretreatment regimen, 32.1 ± 26.0 RU/ml after 2 months of pretreatment, and 24.1 ± 23.1 RU/ml during early pregnancy (P antiphospholipid antibody titers were 52.8 ± 30.7 RU/ml before pretreatment, 38.5 ± 34.2 RU/ml after pretreatment, and 33.9 ± 24.7 RU/ml during early pregnancy; the decrease in antiphospholipid antibodies was lower in the miscarriage group than in the live birth group (P antiphospholipid antibody titers correlated with better pregnancy outcomes. The shorter treatment regimen was effective and economical. PMID:28139508

  7. Construction of a humanized antibody to hepatitis B surface antigen by specificity-determining residues (SDR)-grafting and de-immunization.

    Science.gov (United States)

    Kim, Keun-Soo; Kim, Hyun-Jung; Han, Byung Woo; Myung, Pyung-Keun; Hong, Hyo Jeong

    2010-05-28

    We previously constructed a humanized antibody, HuS10, by grafting the complementarity-determining regions (CDRs) of a parental murine monoclonal antibody into the homologous human antibody sequences. This process is termed CDR grafting. Some residues that were thought to affect the CDR loops and stabilize the structure of the variable regions were retained in the framework region. HuS10 exhibited in vivo virus-neutralizing activity, but its murine content had the potential to elicit immune responses in patients. In this study, to minimize the immunogenic potential of HuS10, we replaced 17 mouse residues in HuS10 with the comparable human residues using specificity-determining residue (SDR)-grafting and de-immunization methods. The resultant humanized antibody, HzS-III, had the same affinity and epitope specificity as HuS10 and had reduced immunogenic potential, as assessed by T-cell epitope analysis. Thus, SDR grafting in combination with de-immunization may be a useful strategy for minimizing the immunogenicity of humanized antibodies. In addition, HzS-III may be a good candidate for immunoprophylaxis of HBV infection. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  8. Production of a monoclonal antibody against serum immunoglobulin M of South American camelids and assessment of its suitability in two immunoassays.

    Science.gov (United States)

    Friedrich, Adrián; Ledesma, Martín; Landone, Ignacio; Ferrari, Alejandro; Leoni, Juliana

    2014-09-01

    A monoclonal antibody (mAb) was produced against immunoglobulin M (IgM) of South American camelids. A single radial immunodiffusion (SRID) assay and a competitive enzyme-linked immunosorbent assay (ELISA) were developed to measure IgM in serum samples. Isotype and specificity of the mAb were assessed. The performance of the SRID assay was preliminarily evaluated in terms of working range, plate stability over a 4-week period, and initial intra- and interassay variation. The concentration of IgM was determined in 55 samples by SRID assay and ELISA, and results were not significantly different by t-test (0.64 ± 0.19 mg/ml for the SRID assay, and 0.58 ± 0.24 mg/ml for ELISA; P = 0.1489). The mAb was shown to be stable over the 4-week evaluation period, and the SRID assay was reproducible when tested in triplicate for intra-assay variability and in quadruplicate for interassay variability, with a percentage coefficient of variation of less than or equal to 5%. Also, the SRID assay proved to be sensitive enough to measure IgM levels in undiluted serum samples, and had a good correlation with ELISA. The current study is intended to submit a preliminary report of a mAb against IgM of South American camelids, and suggest the future potential of the mAb developed for diagnostic application, including use in the SRID assay. © 2014 The Author(s).

  9. [Study of anti-idiotype antibodies to human monoclonal antibody].

    Science.gov (United States)

    Harada, R; Takahashi, N; Owaki, I; Kannagi, R; Endo, N; Morita, N; Inoue, M

    1992-02-01

    A human monoclonal antibody, ll-50 (IgM, lambda), was generated, which reacted specifically with a major of glycolipid present in LS174T colon cancer cells. The glycolipid antigen which reacted with the ll-50 antibody was expected to four sugar residues from its TLC mobility, and it was ascertained that the glycolipid antigen which reacted with ll-50 antibody might be Lc4 antigen [Gal beta 1----3 GLcNAc beta 1----3 Gal beta 1----4 Glc beta 1----1 Cer] judging from TLC immunostaining and ELISA when the reactivity of ll-50 antibody was tested using various pure glycolipids in 3-5 sugar residues as an antigen. Sera in patients with malignant disorders and healthy individuals were analyzed by Sandwich assay of immobilized and biotinylated ll-50 antibody. The serum of the Lc4 antigen recognized by ll-50 antibody was significantly higher in patients with malignant disorders than that in healthy individuals (p less than 0.05). Three mouse monoclonal anti-idiotype antibodies, G3, B3 and C5 (all IgG1), were generated by the immunization of BALB/c mice with ll-50 antibody. These anti-idiotype antibodies specifically bound to to human monoclonal antibody, ll-50 and had a significant inhibitory activity towards the binding of ll-50 antibody to the Lc4 antigen. This indicated that these anti-idiotype antibodies, G3, B3, and C5, were paratope-related anti-idiotype antibodies. G3, B3, and C5 were expected to define the nearest idiotope because they could mutually inhibit ll-50 antibody. Sera in patients with malignant disorders and healthy individuals were analyzed by Sandwich assay of immobilized and biotinylated anti-idiotype antibodies, G3, B3, and C5. As to the ll-50 like antibodies defined by C5 (Id-C5+), the mean serum level in patients with malignant disorders was significantly higher than that in healthy individuals (p less than 0.05). As to the ll-50 like antibodies defined by B3 (Id-B3+), the mean serum level in patients with malignant disorders was significantly higher

  10. Potency determination of inactivated H7 influenza vaccines using monoclonal antibody-based ELISA and biolayer interferometry assays.

    Science.gov (United States)

    Vasudevan, Anupama; Woerner, Amy; Schmeisser, Falko; Verma, Swati; Williams, Ollie; Weir, Jerry P

    2018-03-01

    The single radial immunodiffusion (SRID) assay, the accepted method for determining potency of inactivated influenza vaccines, measures an immunogenic form of the influenza hemagglutinin. Nevertheless, alternative methods for measuring vaccine potency have been explored to address some of the weaknesses of the SRID assay, including limited sensitivity and the requirement for large amounts of standardized reagents. Monoclonal antibody (mAb)-based potency assays also have the ability to detect and measure relevant immunogenic forms of HA. The objective of this study was to continue evaluation of mAb-based alternative methods for measuring the potency of inactivated influenza vaccines, focusing on A(H7N9) pandemic influenza vaccines. Several murine mAbs that recognize different epitopes on the H7 hemagglutinin (HA) were identified and characterized. These mAbs were evaluated in both a mAb-capture ELISA and a mAb-based biolayer interferometry (BLI) assay. Results indicated that potency of inactivated A(H7N9) vaccines, including vaccine samples that were stressed by heat treatment, measured by either alternative method correlated well with potency determined by the traditional SRID potency assay. The availability of multiple H7 mAbs, directed to different HA epitopes, provides needed redundancy in the potency analysis as A(H7N9) viruses continue to evolve antigenically and suggests the importance of having a broad, well-characterized panel of mAbs available for development of vaccines against influenza strains with pandemic potential. In addition, the results highlight the potential of mAb-based platform such as ELISA and BLI for development as alternative methods for determining the potency of inactivated influenza vaccines. Published 2017. This article is a U.S. Government work and is in the public domain in the USA. Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd.

  11. A Study of Anti Beta-2 Glycoprotein I and Anti-Prothrombin Antibodies in Patients with Unexplained Recurrent Pregnancy Losses

    OpenAIRE

    Singh, Angad; Nangia, Anita; Sharma, Sunita; Puri, Manju

    2015-01-01

    To compare the levels of IgG and IgM anti beta-2 glycoprotein I��antibodies and��IgG and IgM anti prothrombin antibodies among women with unexplained recurrent pregnancy losses and women with at least 2 live issues. To compare the prevalence of newer anti beta-2 glycoprotein I & anti prothrombin antibodies with conventional Lupus anticoagulant & anticardiolipin antibodies. 50 women with recurrent pregnancy losses & 50 matched controls were evaluated for the presence of: Lupus anticoagulant���...

  12. The utility of measles and rubella IgM serology in an elimination setting, Ontario, Canada, 2009-2014.

    Science.gov (United States)

    Bolotin, Shelly; Lim, Gillian; Dang, Vica; Crowcroft, Natasha; Gubbay, Jonathan; Mazzulli, Tony; Schabas, Richard

    2017-01-01

    In Canada, measles was eliminated in 1998 and rubella in 2000. Effective measles and rubella surveillance is vital in elimination settings, hinging on reliable laboratory methods. However, low-prevalence settings affect the predictive value of laboratory tests. We conducted an analysis to determine the performance of measles and rubella IgM testing in a jurisdiction where both infections are eliminated. 21,299 test results were extracted from the Public Health Ontario Laboratories database and 1,239 reports were extracted from the Ontario Integrated Public Health Information System (iPHIS) from 2008 and 2010 for measles and rubella, respectively, to 2014. Deterministic linkage resulted in 658 linked measles records (2009-2014) and 189 linked rubella records (2010-2014). Sixty-six iPHIS measles entries were classified as confirmed cases, of which 53 linked to laboratory data. Five iPHIS rubella entries were classified as confirmed, all linked to IgM results. The positive predictive value was 17.4% for measles and 3.6% for rubella. Sensitivity was 79.2% for measles and 100.0% for rubella. Specificity was 65.7% for measles and 25.8% for rubella. Our study confirms that a positive IgM alone does not confirm a measles case in elimination settings. This has important implications for countries that are working towards measles and rubella elimination.

  13. Dyslipidemia and its relationship with antiphospholipid antibodies in APS patients in North Kerala.

    Science.gov (United States)

    Sadanand, Shajit; Paul, Binoy J; Thachil, Emil J; Meletath, Rejadheesh

    2016-12-01

    Antiphospholipid antibody syndrome (APS) is one of the most common acquired thrombophilic disorders resulting in arterial and venous thromboses. APS is a major cause for cerebrovascular accidents or stokes, myocardial infarction, venous thromboembolism and recurrent abortions/pregnancy losses especially in young patients. APS patients have an increased risk of atherosclerotic cardiovascular events. There are only two studies on lipid abnormalities in APS patients. None of them have studied the relationship between individual laboratory tests for APS and lipid profile abnormalities. Here we describe the significance of the relationship between various APS tests and lipid profile abnormalities in a subset of APS patients who presented with arterial thrombosis in a tertiary care hospital. The study was conducted at Government Medical College, which is a tertiary care referral hospital. All patients who presented to the medicine department with APS during a two-year period were studied. A patient was considered to be positive for anticardiolipin (aCL) antibody or anti-β2 glycoprotein (anti-β2G) if the titer was more than 15 IU/mL, and a high titer was considered to be more than 40 IU/mL for Immunoglobulin (lg) IgG and IgM isotypes. The fasting lipid profile was measured in all patients, and lipid profile abnormalities were defined with cutoffs of low-density lipoprotein (LDL) levels of >150 mg/dL, triglyceride (TG) levels of >150 mg/dL, and high-density lipoprotein (HDL) levels of APS, aCL IgG and IgM and anti-β2G IgG and IgM, were determined by statistical analysis. The study population included 77 APS patients, with 53% of patients between 20 and 40 years. The commonest abnormality in the lipid profile test was elevated TG levels of >150 mg/dL in 51.9% of the patients, followed by low HDL levels (150 mg/dL) in 40.2% of the patients. There was a statistically significant relationship between anti-β2G IgG levels and HDL and LDL levels, but not TG levels. Only LDL

  14. Validation of the FAO/IAEA/PANAFTOSA ELISA kit for determination of antibodies against foot-and-mouth disease virus

    International Nuclear Information System (INIS)

    Maradei, E.; Pedemonte, A.

    1998-01-01

    A Liquid phase blocking sandwich ELISA (LPBE) for the detection of foot-and-mouth disease (FMD) antibodies, serotypes O, A and C was validated using sera from bovines free of antibodies and vaccinated bovines. This technique proved to be sensitive and specific for the study of these antibodies. This kit has been prepared by the Pan American Foot-and-Mouth Disease Center (PAHO/WHO) in collaboration with the Animal Production and Health Section of the Joint FAO/IAEA Division, Vienna, Austria and the Institute for Animal Health in Pirbright, United Kingdom. (author)

  15. Prevalence of specific IGM due to toxoplasma, rubella, CMV and c.trachomatis infections during pregnancy

    Directory of Open Access Journals (Sweden)

    Yasodhara P

    2001-01-01

    Full Text Available One hundred and seventy five apparently normal asymptomatic pregnant women were studied prospectively and 247 women with different complications of pregnancy were screened at the time of delivery for infections like Chlamydia trachomatis, Toxoplasma, Rubella and cytomegalovirus (CMV. One hundred and forty two women with normal outcome of pregnancy served as controls. Specific IgM due to these agents were determined in the sera using commercial diagnostic kits. Results of the study showed that chlamydial infection was the most prevalent (29.8% followed by Toxoplasmosis (13.1, Rubella (6.5% and CMV (5.8%. Adverse outcome was seen among those seropositive for Chlamydia, Toxoplasmosis and Rubella. CMV showed no association with adverse outcome of pregnancy. Since Chlamydia and Toxoplasmosis are amenable to treatment with antibiotics, screening for these infections and appropriate treatment would improve outcome of pregnancy.

  16. Detection of serum antitrichomonal antibodies in urogenital trichomoniasis by immunofluorescence.

    Directory of Open Access Journals (Sweden)

    Bhatt R

    1992-04-01

    Full Text Available Trichomonas vaginalis is a frequently encountered genital pathogen in both males and females. In females, vaginitis due to this parasite is one of the most common manifestation. The indirect fluorescent technique (IFA test was carried out to detect antitrichomonal antibodies in 370 female patients using whole cell antigen. Seventy one (19.18% gave positive reaction for either of the class IgG, IgM and IgA antibodies. The level of the IgG class antibodies was found to be higher i.e. 58 (81.69% than IgM 11 (15.27% antibodies, which may be suggestive of past infection or a prolonged manifestation by the organisms.

  17. Antibody capture by mixed-mode chromatography: a comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins.

    Science.gov (United States)

    Pezzini, Jerome; Joucla, Gilles; Gantier, René; Toueille, Magali; Lomenech, Anne-Marie; Le Sénéchal, Caroline; Garbay, Bertrand; Santarelli, Xavier; Cabanne, Charlotte

    2011-11-11

    We evaluated mixed mode chromatography for the capture of recombinant antibodies from CHO cell culture supernatants. We studied PPA HyperCel, HEA HyperCel, MEP HyperCel and Capto adhere resins, which all contain hydrophobic and cationic groups. A microplate approach combined with DoE modeling allowed the exploration of the complex behaviors of these mixed mode resins. Optimal conditions for antibody purification and host cell proteins (HCPs) elimination were determined and then directly up-scaled to laboratory columns. Then we used mass spectrometry to identify the major HCPs potentially coeluted with the antibody. Differences between the four resins in terms of amount, complexity and identity of the HCPs present in the elution fractions were investigated. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. IgM, FcµRs, and malarial immune evasion

    DEFF Research Database (Denmark)

    Czajkowsky, Daniel M; Salanti, Ali; Ditlev, Sisse B

    2010-01-01

    IgM is an ancestral Ab class found in all jawed vertebrates, from sharks to mammals. This ancient ancestry is shared by malaria parasites (genus Plasmodium) that infect all classes of terrestrial vertebrates with whom they coevolved. IgM, the least studied and most enigmatic of the vertebrate Igs...

  19. Characterization of the antibody response to a Haemophilus influenzae type b conjugate vaccine in children with recurrent lower respiratory tract infection

    DEFF Research Database (Denmark)

    Kristensen, K; Barington, T; Pressler, T

    1995-01-01

    once with a Haemophilus influenzae type b (Hib) conjugate vaccine. Total IgG subclasses, total antipolysaccharide Hib antibodies, and antipolysaccharide Hib antibodies of IgM, IgG, IgA, and IgG1-4 specificity were determined by ELISA. There were no significant differences between the two groups in any...... single total IgG subclass, but total IgG measured as the sum of all four subclasses was significantly lower in the children with RLRI than in the controls (P = 0.036). Before vaccination, the children with RLRI had significantly less IgG antipolysaccharide Hib antibody than the controls (P = 0......M response to Hib conjugate vaccine in these children, since this isotype predominates in the primary immune response, i.e., in the absence of immunologic memory.(ABSTRACT TRUNCATED AT 250 WORDS)...

  20. Protein microarray-mediated detection of antienterovirus antibodies in serum.

    Science.gov (United States)

    Zhang, Aiying; Xiu, Bingshui; Zhang, Heqiu; Li, Ning

    2016-04-01

    To utilize prokaryotic gene expression and protein microarray to develop and evaluate a sensitive, accurate protein microarray assay for detecting antienterovirus antibodies in serum samples from patients with hand, foot and mouth disease (HFMD). Enterovirus 71 (EV71) and coxsackievirus A16 (CA16), two common causative agents for HFMD, were used for assay development. Serum was collected from patients with HFMD and healthy controls. EV71 and CA16 VP1 and VP3 genes were expressed in transfected Escherichia coli; the resultant VP1 and 3 proteins were used in a microarray assay for human serum EV71 and CA16 immunoglobulin (Ig) M and IgG. To validate the microarray assay, serum samples were tested for EV71 IgM using enzyme-linked immunosorbent assay (ELISA). Out of 50 patients with HFMD, EV71 IgM and CA16 IgM was detected in 80% and 44% of serum samples, respectively, using protein microarray, and EV71 IgM was detected in 78% of samples using ELISA. Protein microarray and ELISA showed 100% specificity for EV71-IgM detection. The protein microarray assay developed in the present study shows potential as a sensitive technique for detecting EV71 IgM in serum samples from patients with HFMD. © The Author(s) 2016.

  1. Naturally occurring bactericidal antibodies specific for Haemophilus influenzae lipooligosaccharide are present in healthy adult individuals.

    Science.gov (United States)

    Choi, Joshua; Nix, Eli B; Gaultier, Gabrielle N; Cox, Andrew D; McCready, William; Ulanova, Marina

    2015-04-15

    Nontypeable Haemophilus influenzae (NTHi), a typical mucosal pathogen largely responsible for respiratory infections and pediatric otitis media, has been increasingly recognized as a significant cause of invasive disease, especially in immunocompromised individuals. Lipooligosaccharide (LOS) is a conserved molecule with an important role in H. influenzae virulence and immune evasion, and it may be considered as a vaccine candidate. However, abilities of H. influenzae LOS to induce protective immune response are poorly understood. The goal of this study was to determine whether antibodies against LOS isolated from H. influenzae strains Eagan, Rd and NTHi 375 are present in the sera of normal individuals. Antigen specific IgG and IgM were studied in sera of 71 and 30 healthy adults, respectively. IgG specific for LOS of all three strains was ubiquitously present in our sample population while IgM specific for Eagan, Rd and NTHi 375 LOS compounds was detected in 37%, 63%, and 40% of samples, respectively. All tested serum samples exhibited bactericidal activity against all three H. influenzae strains; the removal of anti-LOS antibodies from the sera resulted in significant increases in bacterial survival of the corresponding strain. NTHi 375 exhibited the highest serum resistance, whereas the Rd strain was the least resistant. Serum bactericidal activity of anti-LOS antibody was mediated via the classical complement pathway. These findings suggest that in healthy adults, naturally acquired complement-activating anti-LOS antibodies significantly contribute to the overall serum bactericidal activity against both encapsulated and non-encapsulated strains of H. influenzae. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Prenatal toxoplasmosis antibody and childhood autism.

    Science.gov (United States)

    Spann, Marisa N; Sourander, Andre; Surcel, Heljä-Marja; Hinkka-Yli-Salomäki, Susanna; Brown, Alan S

    2017-05-01

    There is evidence that some maternal infections during the prenatal period are associated with neurodevelopmental disorders, such as childhood autism. However, the association between autism and Toxoplasma gondii (T. gondii), an intracellular parasite, remains unclear. The authors examined whether serologically confirmed maternal antibodies to T. gondii are associated with odds of childhood autism in offspring. The study is based on a nested case-control design of a large national birth cohort (N = 1.2 million) and the national psychiatric registries in Finland. There were 874 cases of childhood autism and controls matched 1:1 on date of birth, sex, birthplace and residence in Finland. Maternal sera were prospectively assayed from a national biobank for T. gondii IgM and IgG antibodies; IgG avidity analyses were also performed. High maternal T. gondii IgM antibody was associated with a significantly decreased odds of childhood autism. Low maternal T. gondii IgG antibody was associated with increased offspring odds of autism. In women with high T. gondii IgM antibodies, the IgG avidity was high for both cases and controls, with the exception of three controls. The findings suggest that the relationship between maternal T. gondii antibodies and odds of childhood autism may be related to the immune response to this pathogen or the overall activation of the immune system. Autism Res 2017, 10: 769-777. © 2016 International Society for Autism Research, Wiley Periodicals, Inc. © 2016 International Society for Autism Research, Wiley Periodicals, Inc.

  3. A New Highly Selective and Specific Anti-puerarin polyclonal Antibody for Determination of Puerarin Using a Mannich Reaction Hapten Conjugate.

    Science.gov (United States)

    Udomsin, Orapin; Krittanai, Supaluk; Kitisripanya, Tharita; Tanaka, Hiroyuki; Putalun, Waraporn

    2018-01-01

    Puerarin (PUE) is a phytoestrogen found in Pueraria candollei and Pueraria lobata. These plants are substantial for traditional medicine in various Asian countries. PUE is a key marker that can be found only in the Pueraria species. To establish the method for determination of PUE content which is required for quality control of pharmaceutical products. PUE-cationized bovine serum albumin conjugate was created via Mannich reaction. After the rabbit immunization, the obtain anti-PUE polyclonal antibody (PAb) was used to develop an enzyme-linked immunosorbent assay (ELISA). An anti-PUE PAb possess a great sensitivity and specificity. The cross-reactivity analysis shows no cross-reaction of an established antibody against other substances. In addition, we successfully developed an indirect competitive ELISA (icELISA) for the quantitative analysis of PUE. The result of method validation conforms to acceptance criteria and correlates with high-performance liquid chromatography, the reference method. The icELISA was applied to determine PUE content in Pueraria spp. plant samples and its derived pharmaceutical products. This highly specific immunogen was created from the Mannich reaction. An icELISA can also be applied to other research propose in the further studies. The new immunogen conjugated (puerarin-cBSA) via Mannich reaction was successfully in rising of antibody against puerarin (PUE)The obtained anti-PUE polyclonal antibody (PAb) was high sensitivity and specificity to PUEAn indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed and validated using anti-PUE PAbThe established icELISA was applied to determine PUE content in various tuberous root of Pueraria sppMoreover, icELISA method can be applicable in Pueraria spp. derived products. Abbreviations used: PUE: Puerarin; PAb: Polyclonal antibody; ELISA: Enzyme-linked immunosorbent assay; icELISA: Indirect competitive ELISA; cBSA: Cationized bovine serum albumin.

  4. Comparison of the C-mediating killing activity and C-activating properties of mouse monoclonal and polyclonal antibodies against Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    T. L. Kipnis

    1992-01-01

    Full Text Available A Mouse polyclonal antiserum against Trypanosoma cruzi or its IgG and IgM fractions and five monoclonal antibodies (two IgM, two IgG1 and one IgG2a recognize and combine with membrane components of trypomastigote forms of the parasite as revealed by immunofluorescence. Although all these antibodies sensitize trypomastigotes and prepare them to activate the complement (C system, as measured by consumption of total C, C4, B and C3, only the polyclonal antiserum or its IgG, IgM and Fabμ fragments were able to induce trypanosome lysis by the alternative C pathway.

  5. Antiphosphatidylserine/prothrombin antibodies (aPS/PT) as potential diagnostic markers and risk predictors of venous thrombosis and obstetric complications in antiphospholipid syndrome.

    Science.gov (United States)

    Shi, Hui; Zheng, Hui; Yin, Yu-Feng; Hu, Qiong-Yi; Teng, Jia-Lin; Sun, Yue; Liu, Hong-Lei; Cheng, Xiao-Bing; Ye, Jun-Na; Su, Yu-Tong; Wu, Xin-Yao; Zhou, Jin-Feng; Norman, Gary L; Gong, Hui-Yun; Shi, Xin-Ming; Peng, Yi-Bing; Wang, Xue-Feng; Yang, Cheng-De

    2018-03-28

    The aim of the study was to determine the prevalence and clinical associations of antiphosphatidylserine/prothrombin antibodies (aPS/PT) with thrombosis and pregnancy loss in Chinese patients with antiphospholipid syndrome (APS) and seronegative APS (SNAPS). One hundred and eighty six Chinese patients with APS (67 primary, 119 secondary), 48 with SNAPS, 176 disease controls (79 systemic lupus erythematosus [SLE], 29 Sjogren's syndrome [SS], 30 ankylosing spondylitis [AS], 38 rheumatoid arthritis [RA]) and 90 healthy donors were examined. IgG and IgM aPS/PT, IgG/IgM/IgA anticardiolipin (aCL) and IgG/IgM/IgA anti-β2-glycoprotein I (anti-β2GPI) antibodies were tested by ELISA. One hundred and sixty (86.0%) of APS patients were positive for at least one aPS/PT isotype. One hundred and thirty five (72.6%) were positive for IgG aPS/PT, 124/186 (66.7%) positive for IgM aPS/PT and 99 (53.2%) positive for both. Approximately half of the SNAPS patients were positive for IgG and/or IgM aPS/PT. Highly significant associations between IgG aPS/PT and venous thrombotic events (odds ratio [OR]=6.72) and IgG/IgM aPS/PT and pregnancy loss (OR=9.44) were found. Levels of IgM aPS/PT were significantly different in APS patients with thrombotic manifestations and those with fetal loss (p=0.014). The association between IgG/IgM aPS/PT and lupus anticoagulant (LAC) was highly significant (pAPS was 101.6. Notably, 91.95% (80/87) of LAC-positive specimens were positive for IgG and/or IgM aPS/PT, suggesting aPS/PT is an effective option when LAC testing is not available. Anti-PS/PT antibody assays demonstrated high diagnostic performance for Chinese patients with APS, detected some APS patients negative for criteria markers and may serve as potential risk predictors for venous thrombosis and obstetric complications.

  6. Comparison of different monoclonal antibodies against immunosuppressive proteins of Ascaris suum

    Directory of Open Access Journals (Sweden)

    T.M. Oshiro

    2004-02-01

    Full Text Available The extract of Ascaris suum suppresses the humoral and cellular immune responses to unrelated antigens in the mouse. In order to further characterize the suppressive components of A. suum, we produced specific monoclonal antibodies which can provide an important tool for the identification of these proteins. The A. suum immunosuppressive fractions isolated by gel filtration from an extract of adult worms were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells and the cloned hybrid cells obtained were screened to determine the specificity of secreted antibodies. Three monoclonal antibodies named MAIP-1, MAIP-2 and MAIP-3 were selected and were shown to react with different epitopes of high molecular weight proteins from the A. suum extract. All antibody molecules have kappa-type light chains but differ in heavy chain isotype. MAIP-1 is a mouse IgM, MAIP-2 is an IgA immunoglobulin and MAIP-3 is an IgG1 immunoglobulin and they recognize the antigen with affinity constants of 1.3 x 10(10 M-1, 7.1 x 10(9 M-1 and 3.8 x 10(7 M-1, respectively. The proteins recognized by these monoclonal antibodies (PAS-1, PAS-2 and PAS-3 were purified from the crude extract by affinity chromatography and injected with ovalbumin in BALB/c mice in order to determine their suppressive activity on heterologous antibody production. It was demonstrated that these three proteins are able to significantly suppress anti-ovalbumin antibody secretion, with PAS-1 being more efficient than the others.

  7. Ki-67 immunoreactivity in meningiomas--determination of the proliferative potential of meningiomas using the monoclonal antibody Ki-67

    DEFF Research Database (Denmark)

    Madsen, C; Schrøder, H D

    1997-01-01

    The proliferative potential of 66 human intracranial meningiomas (15 benign, 15 atypical, 15 recurrent, 13 bone-invasive, and 8 brain-invasive) was investigated by means of immunohisto-chemistry using the monoclonal antibody Ki-67. This antibody recognizes a nuclear antigen present in human cells...... during all active phases of the cell cycle, but absent in the resting phase. The purpose of this retrospective study was to estimate the Ki-67-labelling index (L.I.) and see if this index could help discriminate between the different groups of meningiomas. Our results demonstrated that L.I. could...

  8. IgM multiple myeloma: disease definition, prognosis, and differentiation from Waldenstrom's macroglobulinemia.

    Science.gov (United States)

    Schuster, Steven R; Rajkumar, Sundararajan Vincent; Dispenzieri, Angela; Morice, William; Aspitia, Alvaro Moreno; Ansell, Stephen; Kyle, Robert; Mikhael, Joseph

    2010-11-01

    IgM multiple myeloma (MM) and Waldenstrom's macroglobulinemia (WM) are two distinct hematologic entities with the common finding of an IgM monoclonal gammopathy. Distinguishing these two diagnoses is critical as the approach to therapy is different. A priori, we defined IgM MM as a symptomatic clonal plasma cell proliferative disorder characterized by an IgM monoclonal protein (regardless of size), 10% or more plasma cells on bone marrow biopsy, plus the presence of lytic bone lesions and/or translocation t(11;14). Twenty-one patients met this definition of IgM MM. All 21 patients had lytic bone lesions. Of the 16 evaluated with FISH, 6 (38%) demonstrated t(11;14). Median overall survival was 30 months, which is similar to non-IgM myeloma patients treated during this period and shorter than what would be expected for WM. In this, the largest series of patients with IgM MM, we describe the clinical features and prognosis of patient with IgM MM using a strict definition for the disease. The subset of patients without lytic lesions or t(11;14) but with immunophenotypic features suggestive of MM need further study. © 2010 Wiley-Liss, Inc.

  9. Calorimetric features of IgM gammopathies. Implication for patient’s diagnosis and monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Krumova, Sashka; Todinova, Svetla; Danailova, Avgustina [Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., bl. 21, 1113 Sofia (Bulgaria); Petkova, Violeta; Dimitrova, Keranka; Gartcheva, Lidia [National Specialized Hospital for Active Treating of Haematological Diseases, Sofia 1756 (Bulgaria); Taneva, Stefka G., E-mail: stefka.germanova@ehu.es [Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., bl. 21, 1113 Sofia (Bulgaria); Unidad de Biofísica (CSIC-UPV/EHU) and Departamento de Bioquímica y Biología Molecular, Universidad del País Vasco, 48080 Bilbao (Spain)

    2015-09-10

    Highlights: • DSC reveals distinct calorimetric features for IgM gammopaties. • Calorimetry sets of IgM multiple myeloma and Waldenström’s macroglobulinemia patients. • DSC features correlate with the level of involved IgM heavy/light chains. • DSC and “Heavylite” are complementary methods for patients diagnostics/monitoring. - Abstract: The serum proteome of patients featuring immunoglobulin M (IgM) gammopathy and diagnosed as having IgM multiple myeloma or Waldenström’s macroglobulinemia was characterized by differential scanning calorimetry and HevyLite™ (HLC) assay. The thermodynamic properties of the thermograms and their deviation from the typical healthy thermogram were found to correlate with the monoclonal protein concentration and the level of the involved (monoclonal) IgM heavy/light chains. Patients monitoring during treatment showed that the variations in the shape of the DSC profiles corresponded to fluctuations of the IgM heavy/light chain levels verifying the two parameters as non-invasive markers for disease progression.

  10. Highly sensitive determination of diclofenac based on resin beads and a novel polyclonal antibody by using flow injection chemiluminescence competitive immunoassay

    Science.gov (United States)

    Shi, Jing; Xu, Mingxia; Tang, Qinghui; Zhao, Kang; Deng, Anping; Li, Jianguo

    2018-02-01

    A novel flow injection chemiluminescence immunoassay for simple, sensitive and low-cost detection of diclofenac was established based on specific binding of antigen and antibody. Carboxylic resin beads used as solid phase carrier materials provided good biocompatibility and large surface-to-volume ratio for modifying more coating antigen. There was a competitive process between the diclofenac in solution and the immobilized coating antigen to react with the limited binding sites of the polyclonal antibody to form the immunocomplex. The second antibody labelled with horseradish peroxidase was introduced into the immunosensor and trapped by captured polyclonal antibody against diclofenac, which could effectively amplify chemiluminescence signals of luminol-PIP-H2O2. Under optimal conditions, the diclofenac could be detected quantitatively. The chemiluminescence intensity decreased linearly with the logarithm of the diclofenac concentration in the range of 0.1-100 ng mL- 1 with a detection limit of 0.05 ng mL- 1 at a signal-to-noise ratio of 3. The immunosensor exhibited high sensitivity, specificity and acceptable stability. This easy-operated and cost-effective analytical method could be valuable for the diclofenac determination in real water samples.

  11. Determination of auto-antibodies to native and oxidized low-density lipoproteins (LDL) in serum of patients underwent coronariography in the Medical-Surgical Research Center (MSRC)

    International Nuclear Information System (INIS)

    Conde CerdeiraI, Hector; Soto Lopez, Yosdel; Aroche Aportela, Ronald

    2010-01-01

    Low-density lipoprotein (LDL) oxidation is an important event in atherosclerosis development. The relationship between oxidized LDL (oxLDL) autoantibodies and coronary artery disease (CAD) remains controversial. IgM and IgG autoantibodies to oxLDL were measured in twenty patients undergoing clinically indicated coronary angiography, and in ten young healthy volunteers from the Center of Molecular Immunology. The levels of IgM autoantibodies to oxLDL did not differ between no CAD patients and healthy subjects, but the levels of IgM autoantibodies to oxLDL of these two groups were higher compared with the one of CAD patient group. Our results, although preliminary, supports the hypothesis that this kind of Abs might be inversely associated with the presence of atherosclerosis

  12. The Biological Function of Antibodies Induced by the RTS,S/AS01 Malaria Vaccine Candidate is Determined by Their Fine Specificity

    Science.gov (United States)

    2016-05-31

    malaria vaccine candidate is determined by their fine specificity Sidhartha Chaudhury1, Christian F. Ockenhouse2, Jason A. Regules3, Sheetij Dutta4...Anders Wallqvist1, Erik Jongert5, Norman C. Waters4, Franck Lemiale2 and Elke Bergmann‑Leitner4* Abstract Background: Recent vaccine studies have shown... vaccine candidate, RTS,S, have shown that circumsporozoite protein (CSP)‑specific antibodies and CD4+ T cell responses are associated with protection

  13. Inefficient binding of IgM immune complexes to erythrocyte C3b-C4b receptors (CR1) and weak incorporation of C3b-iC3b into the complexes

    DEFF Research Database (Denmark)

    Kávai, M; Rasmussen, J M; Baatrup, G

    1988-01-01

    The binding of soluble complement-reacted IgM immune complexes (IC) to erythrocyte (E) C3b-C4b receptors (CR1) and the incorporation of C3b-iC3b into solid phase IgM-IC was investigated. The optimal binding of liquid phase IgM-IC to E-CR1 was obtained with IC formed at moderate antibody excess...

  14. The future of monoclonal antibody technology

    OpenAIRE

    Zider, Alexander; Drakeman, Donald L

    2010-01-01

    With the rapid growth of monoclonal antibody-based products, new technologies have emerged for creating modified forms of antibodies, including fragments, conjugates and multi-specific antibodies. We created a database of 450 therapeutic antibodies in development to determine which technologies and indications will constitute the “next generation” of antibody products. We conclude that the antibodies of the future will closely resemble the antibodies that have already been approved for commer...

  15. Thyroglobulin (Tg) recovery testing with quantitative Tg antibody measurement for determining interference in serum Tg assays in differentiated thyroid carcinoma

    NARCIS (Netherlands)

    Persoon, ACM; Links, TP; Wilde, J; Sluiter, WJ; Wolffenbuttel, BHR; van den Ouweland, JMW

    Background: Thyroglobulin (Tg) measurements are complicated by interference from Tg autoantibodies (TgAbs) or heterophilic antibodies (HAMAs). We used a new automated immunochemiluminometric assay (ICMA) with Tg recovery (TgR) on the Nichols Advantage (R) platform to reassess the clinical utility of

  16. Assessment of the diagnostic performance of the IDS-iSYS tests for toxo IgG, toxo IgM and avidity.

    Science.gov (United States)

    Levigne, Pauline; Peyron, François; Wallon, Martine

    2016-10-01

    When acquired during pregnancy toxoplasmosis can have devastating consequences on the fetus. As maternal infection is in the majority of cases subclinical, the diagnosis of toxoplasmosis relies on serological tests for the detection of IgG, IGM and the mesure of IgG avidity. We evaluated the performance of IDS-iSYS a new automatized instrument based on chemiluminescence for the diagnosis of the disease. Our study was based on non-selected samples received in our laboratory either for the determination of serological status or for distinguishing acute from chronic infection. Seven hundred eighty three samples were enrolled in the study. Compared with Architect IgG and IgM assays, the sensitivity and specificity were respectively 99% and 99% for IgG, and 75% and 97% for IgM. We observed higher remaining titers for IDS iSYS IgG, which could reduce the proportions of patients who have to be retested because of doubtful titers. IgM detection and avidity scored equivalent performance with both methods. This automate appears to be a reliable and easy-to-use tool for diagnosing toxoplasmosis in different clinical settings. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. A Study of Anti Beta-2 Glycoprotein I and Anti-Prothrombin Antibodies in Patients with Unexplained Recurrent Pregnancy Losses.

    Science.gov (United States)

    Singh, Angad; Nangia, Anita; Sharma, Sunita; Puri, Manju

    2016-06-01

    To compare the levels of IgG and IgM anti beta-2 glycoprotein I antibodies and IgG and IgM anti prothrombin antibodies among women with unexplained recurrent pregnancy losses and women with at least 2 live issues. To compare the prevalence of newer anti beta-2 glycoprotein I & anti prothrombin antibodies with conventional Lupus anticoagulant & anticardiolipin antibodies. 50 women with recurrent pregnancy losses & 50 matched controls were evaluated for the presence of: Lupus anticoagulant-screened by LA sensitive aPTT& DRVV and confirmatory Staclot Assay. ELISA kits were used for detecting IgG & IgM anticardiolipin, anti beta-2 glycoprotein I & anti prothrombin antibodies. 11/50 (22 %) women in study group and none in control group had circulating antiphospholipid antibodies. 2 cases (4 %) had lupus anticoagulant. 1 case (2 %) had anticardiolipin antibody & 6 cases (12 %) were positive for anti beta-2 Glycoprotein I antibody (p value = 0.027). 3 cases (6 %) had anti prothrombin antibody. All were mutually exclusive except for one. Women with recurrent pregnancy losses should be tested for anti beta-2 Glycoprotein I antibodies & anti prothrombin antibodies in addition to conventional lupus anticoagulant and anticardiolipin antibodies. This approach can decrease the incidence of SNAP (seronegative antiphospholipid syndrome) cases while establishing the true prevalence of antiphospholipid syndrome.

  18. Selective Depletion of Neuropathy-Related Antibodies from Human Serum by Monolithic Affinity Columns Containing Ganglioside Mimics

    NARCIS (Netherlands)

    Tetala, Kishore K. R.; Heikema, Astrid P.; Pukin, Aliaksei V.; Weijers, Carel A. G. M.; Tio-Gillen, Anne P.; Gilbert, Michel; Endtz, Hubert P.; van Belkum, Alex; Zuilhof, Han; Visser, Gerben M.; Jacobs, Bart C.; van Beek, Teris A.

    2011-01-01

    Monolithic columns containing ganglioside GM2 and GM3 mimics were prepared for selective removal of serum anti-ganglioside antibodies from patients with acute and chronic immune-mediated neuropathies. ELISA results demonstrated that anti-GM2 IgM antibodies in human sera and a mouse monoclonal

  19. Antiphosphatidylserine/prothrombin antibodies (aPS/PT) as potential markers of antiphospholipid syndrome.

    Science.gov (United States)

    Vlagea, Alexandru; Gil, Antonio; Cuesta, Maria V; Arribas, Florencia; Diez, Jesús; Lavilla, Paz; Pascual-Salcedo, Dora

    2013-06-01

    The antiphospholipid antibodies present in antiphospholipid syndrome (APS) are directed at a number of phospholipid-binding proteins: β2 glycoprotein I (β2GPI), prothrombin, and so on. Antibodies directed at β2GPI are accepted as a classification criterion for APS, while the presence of antiprothrombin antibodies is not. In the present article, we investigated the possible role of antiphosphatidylserine/prothrombin antibodies (aPS/PT) as marker of APS on a cohort of 295 individuals with APS (95 primary APS and 45 secondary APS) and APS-related diseases. We found aPS/PT to be highly associated with venous thrombosis (immunoglobulin G [IgG] aPS/PT odds ratio [OR], 7.44; 95% confidence interval [CI], 3.97-13.92 and IgM aPS/PT OR, 2.54; 95% CI, 1.35-4.77) and obstetric abnormalities (IgG aPS/PT OR, 2.37; 95% CI, 1.04-5.43), but not with arterial thrombosis. A very high degree of concordance between the concentration of aPS/PT and lupus anticoagulant activity was demonstrated. Therefore, we support the inclusion of aPS/PT determination as second-level assay to confirm APS classification.

  20. Seroepidemiological Survey for Coxiella burnetii Antibodies and Associated Risk Factors in Dutch Livestock Veterinarians

    Science.gov (United States)

    Van den Brom, René; Schimmer, Barbara; Schneeberger, Peter M.; Swart, Wim A.; van der Hoek, Wim; Vellema, Piet

    2013-01-01

    Since 2007, Q fever has become a major public health problem in the Netherlands and goats were the most likely source of the human outbreaks in 2007, 2008 and 2009. Little was known about the consequences of these outbreaks for those professional care providers directly involved. The aim of this survey was to estimate the seroprevalence of antibodies against C. burnetii among Dutch livestock veterinarians and to determine possible risk factors. Single blood samples from 189 veterinarians, including veterinary students in their final year, were collected at a veterinary conference and a questionnaire was filled in by each participant. The blood samples were screened for IgG antibodies against phase I and phase II antigen of C. burnetii using an indirect immunofluorescent assay, and for IgM antibodies using an ELISA. Antibodies against C. burnetii were detected in 123 (65.1%) out of 189 veterinarians. Independent risk factors associated with seropositivity were number of hours with animal contact per week, number of years graduated as veterinarian, rural or sub urban living area, being a practicing veterinarian, and occupational contact with swine. Livestock veterinarians should be aware of this risk to acquire an infection with C. burnetii. Physicians should consider potential infection with C. burnetii when treating occupational risk groups, bearing in mind that the burden of disease among veterinarians remains uncertain. Vaccination of occupational risk groups should be debated. PMID:23342063

  1. Seroepidemiological survey for Coxiella burnetii antibodies and associated risk factors in Dutch livestock veterinarians.

    Science.gov (United States)

    Van den Brom, René; Schimmer, Barbara; Schneeberger, Peter M; Swart, Wim A; van der Hoek, Wim; Vellema, Piet

    2013-01-01

    Since 2007, Q fever has become a major public health problem in the Netherlands and goats were the most likely source of the human outbreaks in 2007, 2008 and 2009. Little was known about the consequences of these outbreaks for those professional care providers directly involved. The aim of this survey was to estimate the seroprevalence of antibodies against C. burnetii among Dutch livestock veterinarians and to determine possible risk factors. Single blood samples from 189 veterinarians, including veterinary students in their final year, were collected at a veterinary conference and a questionnaire was filled in by each participant. The blood samples were screened for IgG antibodies against phase I and phase II antigen of C. burnetii using an indirect immunofluorescent assay, and for IgM antibodies using an ELISA. Antibodies against C. burnetii were detected in 123 (65.1%) out of 189 veterinarians. Independent risk factors associated with seropositivity were number of hours with animal contact per week, number of years graduated as veterinarian, rural or sub urban living area, being a practicing veterinarian, and occupational contact with swine. Livestock veterinarians should be aware of this risk to acquire an infection with C. burnetii. Physicians should consider potential infection with C. burnetii when treating occupational risk groups, bearing in mind that the burden of disease among veterinarians remains uncertain. Vaccination of occupational risk groups should be debated.

  2. Structural Characterization and Determinants of Specificity of Single- Chain Antibody Inhibitors of Membrane-Type Serine Protease 1

    Science.gov (United States)

    2007-03-01

    protease involved in male chromatin remodeling blocks the development of sea urchin embryos at the initial cell cycle. J. Cell Biochem. 98, 335–342. 18...Macrophage Morphology Changes and Inhibition of Nitric Oxide Production by Macrophages. The cleavage of MSP-1 by MT-SP1 was then tested in primary cells in...inhibitor (Fig. 3) were studied. The morphology change in response to MSP-1 was independent of HAI-1 or anti-MT-SP1 antibody presence. Both inhibitors

  3. Symptomatic Secondary Selective IgM Immunodeficiency in Adult Man with Undiagnosed Celiac Disease

    OpenAIRE

    Magen, Eli; Feldman, Viktor; Joseph, Mishal; Israel, Hadari

    2012-01-01

    Selective IgM immunodeficiency (SIgMID) is a heterogeneous disorder with no known genetic background and may occur as a primary or a secondary condition. Celiac disease has been reported in association with several humeral immunodeficiencies, including isolated severe selective IgA deficiency, panhypogammaglobulinemia, and isolated combined IgA and IgM deficiency. There are only few reported cases of pediatric and adult patients with SIgMID and celiac disease. In this paper, we describe an ad...

  4. Study of the titers of Anti-Epstein-Barr virus antibodies in the sera of atomic bomb survivors

    International Nuclear Information System (INIS)

    Akiyama, Mitoshi; Kusunoki, Yoichiro; Kyoizumi, Seishi; Ozaki, Kyoko; Mizuno, Shoichi; Cologne, J.B.

    1993-01-01

    Antibody titers to Epstein-Barr virus antigens were determined in the sera of 372 atomic bomb survivors to evaluate the effect of the previous radiation exposure on immune competence against the latent infection of the virus. The proportion of persons with high titers (≥ 1:40) of IgG antibodies to the early antigen was significantly elevated in the exposed survivors. Furthermore, the distribution of IgM titers against the viral capsid antigen was significantly affected by radiation dose with an increased occurrence of titers of 1:5 and 1:10 in the exposed persons, although the dose effect was only marginally suggestive when persons with rheumatoid factor were eliminated from the analysis. These results suggest that reactivation of Epstein-Barr virus in the latent stage occurs more frequently in the survivors, even though this might not be affected by the radiation dose. Otherwise, there was neither an increased trend in the prevalence of high titers (≥ 1:640) of IgG antibodies to the viral capsid antigen among the exposed people nor a correlation between the radiation exposure and distributions of titers of IgA antibodies to the viral capsid antigen or antibodies to the anti-Epstein-Barr virus-associated nuclear antigen. (author)

  5. The detection and specifity of class specific antibodies to whole bacteria cells using a solid phase radioimmunoassay

    International Nuclear Information System (INIS)

    Czerkinsky, C.; Rees, A.S.; Bergimeier, L.A.; Challacombe, S.J.

    1983-01-01

    A solid phase radioimmunoassay has been developed which can be used for the detection of isotype specific antibodies to whole bacteria and other particulate antigens, and is applicable to a variety of species. Bacteria are bound to the solid phase by the use either of antibodies, or of methyl glyoxal. Both methods result in a sensitive and reproducible assay, and bacteria do not appear to desorb from the solid phase. The specificity of antibodies to whole bacteria was examined by absorption of antisera with various species of bacteria and retesting, or by determining the binding of antisera to various bacteria bound to the solid phase. Both methods revealed specificity for the bacteria examined. Inhibition studies showed that antibodies to Streptococcus mutans whole cells could be inhibited by purified cell surface antigens glucosyltransferase and antigen I/II, but only minimally by lipoteichoic acid, c polysaccharide or dextran. In murine antisera antibodies of the IgG, IgM, and IgA classes could be detected at amounts of less than 1 ng/ml. (author)

  6. [Diagnosis of rubella by demonstrating rubella-specific 19 S and 7 S antibodies (author's transl)].

    Science.gov (United States)

    Dibbert, H J

    1976-03-01

    Identification and measurement of rubella antibody of 19 S (IgM) and 7 S (IgG) immunoglobulins by sucrose density gradient centrifugation and a modified hemagglutination-inhibition (HAI)-test is described. Human group 0 of red blood cells were employed after trypsination. The method proved to be highly reliable and sensitive for the rapid detection of rubella IgM antibody in routine research to make an accurate diagnosis of recent rubella infection. Sera from 185 pregnant women, 200 children suspect of congenital infection, 15 mothers of intrauterin infected children, and 32 cases of rubella vaccine-induced immunity control were studied. In postnatal rubella infections, specific IgM-antibody could be detected in all sera after the onset of the rash up to eight or ten weeks after infection. From March 1973 until September 1974 laboratory diagnosis confirmed 30 of 185 pregnant women to have been recently infected with rubella. In less than 3% (4 cases) the results could not confirm or exclude a recent history of rubella. Among 200 children with congenital defects or some irregular development, specific IgM antibody was detected in 36 cases. The diagnosis of congenital rubella was confirmed or detected in the first months of life. One of the infected children was well at birth but later developed mental retardation and hearing loss, and some others showed only minor defects. In some children rubell IgM antibody persisted 5-14 months. IgM rubella antibody was not found in the sera of 15 mothers from children with intrauterine infection confirmed by IgM antibody demonstration. Post-vaccination-tests showed serum IgM and IgG immunoglobulin-responses to be similar to the natural rubella-virus-infection. Maximal titers of both immunoglobulin-fractions were but four-fold lower after vaccination (28 of 32 tested cases). The described method is able to verify the time of infection by testing a single serum. The HAI-test of the 12 fractions collected from each serum after

  7. Autoimmunity related to IgM monoclonal gammopathy of undetermined significance. Peripheral neuropathy and connective tissue sensibilization caused by IgM M-proteins

    DEFF Research Database (Denmark)

    Jønsson, V; Schrøder, H D; Nolsøe, C

    1988-01-01

    In eight of 10 consecutive cases of IgM monoclonal gammopathy of undetermined significance (MGUS), the M-protein had specificity towards various tissues as estimated by direct and indirect immunofluorescence studies of skin and/or sural nerve biopsies. Five of the cases had neuropathy. In three o...

  8. ELISA to measure neutralizing capacity of anti-C1-inhibitor antibodies in plasma of angioedema patients.

    Science.gov (United States)

    Engel, Ruchira; Rensink, Irma; Roem, Dorina; Brouwer, Mieke; Kalei, Asma; Perry, Dawn; Zeerleder, Sacha; Wouters, Diana; Hamann, Dörte

    2015-11-01

    Neutralizing autoantibodies (NAbs) against plasma serpin C1-inhibitor (C1-inh) are implicated in the rare disorder, acquired angioedema (AAE). There is insufficient understanding of the process of antibody formation and its correlation with disease progression and severity. We have developed an ELISA for detecting neutralizing capacity of anti-C1-inh positive plasma samples that can be used to study changes in NAb repertoire in patient plasma over the course of disease. The ELISA is based on the specific interaction of active C1-inh with its target protease C1s. Decrease in the amount of C1s bound to immobilized C1-inh in the presence of test samples is proportional to the neutralizing capacity of the sample. Assay specificity, intra- and inter-assay variation and assay cut-off are determined using anti-C1-inh antibodies. Assay capability is demonstrated using plasma samples from AAE patients. The assay is specific to a neutralizing anti-C1-inh antibody and shows no interference by a non-neutralizing anti-C1-inh antibody or by the plasma matrix. Intra-assay and inter-assay variations are determined as 17 and 18% respectively. Neutralizing capacity of antibody positive AAE patient plasma samples (n=16) with IgG or IgM type antibodies is readily determined. All samples show positive neutralizing capacity. We have developed a robust, specific and semi-quantitative assay to detect the neutralizing capacity of plasma samples containing anti-C1-inh antibodies. This assay can be an important tool for the study of clinical implications of anti-C1-inh NAbs. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Compositions, antibodies, asthma diagnosis methods, and methods for preparing antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hongjun; Zangar, Richard C.

    2017-01-17

    Methods for preparing an antibody are provided with the method including incorporating 3-bromo-4-hydroxy-benzoic acid into a protein to form an antigen, immunizing a mammalian host with the antigen, and recovering an antibody having an affinity for the antigen from the host. Antibodies having a binding affinity for a monohalotyrosine are provided as well as composition comprising an antibody bound with monohalotyrosine. Compositions comprising a protein having a 3-bromo-4-hydroxy-benzoic acid moiety are also provided. Methods for evaluating the severity of asthma are provide with the methods including analyzing sputum of a patient using an antibody having a binding affinity for monohalotyrosine, and measuring the amount of antibody bound to protein. Methods for determining eosinophil activity in bodily fluid are also provided with the methods including exposing bodily fluid to an antibody having a binding affinity for monohalotyrosine, and measuring the amount of bound antibody to determine the eosinophil activity.

  10. Study of chronic hemolytic anaemia patients in Rio de Janeiro: prevalence of anti-human parvovirus B19 IgG antibodies and the development aplastic crises.

    Science.gov (United States)

    Sant'Anna, Anadayr L M; Garcia, Rita de Cássia N Cubel; Marzoche, Mônica; da Rocha, Heloisa Helena A Gallo; Paula, Maria Tereza M; Lobo, Clarisse C; Nascimento, Jussara P

    2002-01-01

    The prevalence of anti-human parvovirus B19 IgG antibodies was determined in sera from 165 chronic hemolytic anemia patients, receiving medical care at Instituto Estadual de Hematologia (IEHE), Rio de Janeiro, during the year of 1994. This sample represents around 10% of the chronic hemolytic anemia patients attending at IEHE. Most of these patients (140) have sickle cell disease. Anti-B19 IgG antibodies were detected in 32.1% of patients. No statistically significant difference (p > 0.05) was seen between IgG antibody prevalence in male (27.8%) and female (35.5%) patients. Anti-B19 IgG antibodies were more frequent in older (37.6%) than younger (28.2%) than 20 years old patients, although this difference had no statistical significance (p > 0.05). Anti-B19 IgG antibody prevalence showed that 67.9% of patients enrolled in the study were susceptible to B19 acute infection. With the aim to detect acute B19 infection, patients follow up continued until February 1996. During this period four patients presented transient aplastic crisis due to human parvovirus B19 as confirmed by the detection of specific IgM antibodies. All four patients were younger than 20 years old, and 3 were younger than 10 years old. Three of them were sickle cell disease patients. Three of the four acute B19 infection occurred during 1994 springtime.

  11. Study of chronic hemolytic anaemia patients in Rio de Janeiro: prevalence of anti-human parvovirus B19 IgG antibodies and the developement aplastic crises

    Directory of Open Access Journals (Sweden)

    SANT'ANNA Anadayr L.M.

    2002-01-01

    Full Text Available The prevalence of anti-human parvovirus B19 IgG antibodies was determined in sera from 165 chronic hemolytic anemia patients, receiving medical care at Instituto Estadual de Hematologia (IEHE, Rio de Janeiro, during the year of 1994. This sample represents around 10% of the chronic hemolytic anemia patients attending at IEHE. Most of these patients (140 have sickle cell disease. Anti-B19 IgG antibodies were detected in 32.1% of patients. No statistically significant difference (p > 0.05 was seen between IgG antibody prevalence in male (27.8% and female (35.5% patients. Anti-B19 IgG antibodies were more frequent in older (37.6% than younger (28.2% than 20 years old patients, although this difference had no statistical significance (p > 0.05. Anti-B19 IgG antibody prevalence showed that 67.9% of patients enrolled in the study were susceptible to B19 acute infection. With the aim to detect acute B19 infection, patients follow up continued until February 1996. During this period four patients presented transient aplastic crisis due to human parvovirus B19 as confirmed by the detection of specific IgM antibodies. All four patients were younger than 20 years old, and 3 were younger than 10 years old. Three of them were sickle cell disease patients. Three of the four acute B19 infection occurred during 1994 springtime.

  12. Epitope Predictions Indicate the Presence of Two Distinct Types of Epitope-Antibody-Reactivities Determined by Epitope Profiling of Intravenous Immunoglobulins

    Science.gov (United States)

    Luštrek, Mitja; Lorenz, Peter; Kreutzer, Michael; Qian, Zilliang; Steinbeck, Felix; Wu, Di; Born, Nadine; Ziems, Bjoern; Hecker, Michael; Blank, Miri; Shoenfeld, Yehuda; Cao, Zhiwei; Glocker, Michael O.; Li, Yixue; Fuellen, Georg; Thiesen, Hans-Jürgen

    2013-01-01

    Epitope-antibody-reactivities (EAR) of intravenous immunoglobulins (IVIGs) determined for 75,534 peptides by microarray analysis demonstrate that roughly 9% of peptides derived from 870 different human protein sequences react with antibodies present in IVIG. Computational prediction of linear B cell epitopes was conducted using machine learning with an ensemble of classifiers in combination with position weight matrix (PWM) analysis. Machine learning slightly outperformed PWM with area under the curve (AUC) of 0.884 vs. 0.849. Two different types of epitope-antibody recognition-modes (Type I EAR and Type II EAR) were found. Peptides of Type I EAR are high in tyrosine, tryptophan and phenylalanine, and low in asparagine, glutamine and glutamic acid residues, whereas for peptides of Type II EAR it is the other way around. Representative crystal structures present in the Protein Data Bank (PDB) of Type I EAR are PDB 1TZI and PDB 2DD8, while PDB 2FD6 and 2J4W are typical for Type II EAR. Type I EAR peptides share predicted propensities for being presented by MHC class I and class II complexes. The latter interaction possibly favors T cell-dependent antibody responses including IgG class switching. Peptides of Type II EAR are predicted not to be preferentially presented by MHC complexes, thus implying the involvement of T cell-independent IgG class switch mechanisms. The high extent of IgG immunoglobulin reactivity with human peptides implies that circulating IgG molecules are prone to bind to human protein/peptide structures under non-pathological, non-inflammatory conditions. A webserver for predicting EAR of peptide sequences is available at www.sysmed-immun.eu/EAR. PMID:24244326

  13. Epitope predictions indicate the presence of two distinct types of epitope-antibody-reactivities determined by epitope profiling of intravenous immunoglobulins.

    Directory of Open Access Journals (Sweden)

    Mitja Luštrek

    Full Text Available Epitope-antibody-reactivities (EAR of intravenous immunoglobulins (IVIGs determined for 75,534 peptides by microarray analysis demonstrate that roughly 9% of peptides derived from 870 different human protein sequences react with antibodies present in IVIG. Computational prediction of linear B cell epitopes was conducted using machine learning with an ensemble of classifiers in combination with position weight matrix (PWM analysis. Machine learning slightly outperformed PWM with area under the curve (AUC of 0.884 vs. 0.849. Two different types of epitope-antibody recognition-modes (Type I EAR and Type II EAR were found. Peptides of Type I EAR are high in tyrosine, tryptophan and phenylalanine, and low in asparagine, glutamine and glutamic acid residues, whereas for peptides of Type II EAR it is the other way around. Representative crystal structures present in the Protein Data Bank (PDB of Type I EAR are PDB 1TZI and PDB 2DD8, while PDB 2FD6 and 2J4W are typical for Type II EAR. Type I EAR peptides share predicted propensities for being presented by MHC class I and class II complexes. The latter interaction possibly favors T cell-dependent antibody responses including IgG class switching. Peptides of Type II EAR are predicted not to be preferentially presented by MHC complexes, thus implying the involvement of T cell-independent IgG class switch mechanisms. The high extent of IgG immunoglobulin reactivity with human peptides implies that circulating IgG molecules are prone to bind to human protein/peptide structures under non-pathological, non-inflammatory conditions. A webserver for predicting EAR of peptide sequences is available at www.sysmed-immun.eu/EAR.

  14. In-vitro inhibition of IFNγ+ iTreg mediated by monoclonal antibodies against cell surface determinants essential for iTreg function

    Directory of Open Access Journals (Sweden)

    Daniel Volker

    2012-08-01

    Full Text Available Abstract Background IFNγ-producing CD4+CD25+Foxp3+ PBL represent a subtype of iTreg that are associated with good long-term graft outcome in renal transplant recipients and suppress alloresponses in-vitro. To study the mechanism of immunosuppression, we attempted to block cell surface receptors and thereby inhibited the function of this iTreg subset in-vitro using monoclonal antibodies and recombinant proteins. Methods PBL of healthy control individuals were stimulated polyclonally in-vitro in the presence of monoclonal antibodies or recombinant proteins against/of CD178, CD152, CD279, CD28, CD95, and HLA-DR. Induction of IFNγ+ iTreg and proliferation of effector cells was determined using four-color fluorescence flow cytometry. Blockade of iTreg function was analyzed using polyclonally stimulated co-cultures with separated CD4+CD25+CD127-IFNγ+ PBL. Results High monoclonal antibody concentrations inhibited the induction of CD4+CD25+Foxp3+IFNγ+ PBL (anti-CD152, anti-CD279, anti-CD95: p +CD25+CD127-IFNγ+ PBL (anti-CD178, anti-CD152, anti-CD279, anti-CD95: p +CD25+Foxp3+IFNγ+ PBL (rCD152 and rCD95: p +CD25+CD127-IFNγ+ PBL showed lower cell proliferation than co-cultures with CD4+CD25+CD127-IFNγ- PBL (p +CD25+CD127-IFNγ- PBL-containing co-cultures in the presence of monoclonal antibody (anti-CD28, anti-CD152, anti-CD279: p +CD25+CD127-IFNγ+ PBL (with the exception anti-CD28 monoclonal antibody: p +CD25+CD127-IFNγ- PBL but do not efficiently block suppressive iTreg function in co-cultures with CD4+CD25+CD127-IFNγ+ PBL. Conclusions CD178, CD152, CD279, CD28, CD95, and HLA-DR determinants are important for induction and suppressive function of IFNγ+ iTreg.

  15. [Biotic and abiotic determinants of seroprevalence of antibodies against Trypanosoma cruzi in Palmar de Bravo, Puebla, Mexico].

    Science.gov (United States)

    Sosa-Jurado, Francisca; Zumaquero-Ríos, José Lino; Reyes, Pedro A; Cruz-García, Abel; Guzmán-Bracho, Carmen; Monteón, Victor M

    2004-01-01

    To establish the relationship between seroprevalence for antibodies against Trypanosoma cruzi and its relationship with biotic and abiotic factors. A cross-sectional study was conducted between August 2000 and September 2001. The study population consisted of a simple random sample of 390 volunteers residing in Palmar de Bravo, Puebla, Mexico. Sample and data collection procedures included assaying antibodies against T. cruzi with validated assays, and searching for domestic reservoirs and triatomine bugs. The relationship between biotic and abiotic factors with seropositivity was assessed. Statistical analysis was conducted using Kappa values for diagnostic tests; statistical significance was assessed with 2 x 2 tables, chi-squared test with Yates' correction, Fisher exact test, and odds ratios. The seroprevalence of T. cruzi infection in humans was 4%; in domestic reservoirs (horses, pigs, and dogs) only 10% of canine reservoirs were positive. Vector species recognized were T. borberi and T. pallidipennis, with a Dispersion Area Index and a Colonization Index of 55% and 40%, respectively. The most important risk factors associated with positive serology were altitude (>2,150 and <2,180 meters above sea level), presence of triatomines, age, time of residence, and participation in a social assistance program. T. cruzi infection was identified in human beings, vectors, and possibly in domestic reservoirs, in communities located over 2,000 meters above sea level.

  16. Preparation Of Liquid Phase-Double Antibodies Radioimmunoassay For The In Vitro Determination Of Prolactin Hormone In Human Serum

    International Nuclear Information System (INIS)

    MEHANY, N.L.; EL-KOLALY, M.T.; EBEID, N.H.; MEKY, N.H.

    2009-01-01

    In the present study, the preparation of the basic reagents of prolactin (PRL) radioimmunoassay (RIA) technique using liquid phase double antibody with low cost is considered to be the main objective. Three primary components were prepared and characterized to obtain valid and accurate system. These components were polyclonal antibody (anti-PRL), 125 I-prolactin ( 125 I-PRL) radio-iodinated tracer and PRL standards. The production of polyclonal anti-PRL was undertaken by immunizing eight males of white New-Zealand rabbits (two groups) with highly purified PRL antigen through primary injection and five booster doses subcutaneously and intramuscular. The preparation of radio-iodinated ( 1 '2 5 I-PRL) tracer was carried out using chloramine-T method. The preparation of PRL standards were carried out using highly purified PRL antigen in assay buffer. The obtained PRL-antisera were characterized in terms of titer, immuno response and displacement profile. Formulation, optimization and validation of the local liquid phase RIA system were carried out. The results obtained provide a highly sensitive, specific and accurate RIA system of PRL. In conclusion, this technique could be used in diagnosis of pituitary dysfunction such as hyperprolactinaemia and hyperprolactinaemia, prolactinoma, galactorrhoea, amenorrhea and diagnosis of infertility in males and females.

  17. Two distinct subtypes of hepatitis B virus-related acute liver failure are separable by quantitative serum immunoglobulin M anti-hepatitis B core antibody and hepatitis B virus DNA levels

    DEFF Research Database (Denmark)

    Dao, Doan Y; Hynan, Linda S; Yuan, He-Jun

    2012-01-01

    Hepatitis B virus (HBV)-related acute liver failure (HBV-ALF) may occur after acute HBV infection (AHBV-ALF) or during an exacerbation of chronic HBV infection (CHBV-ALF). Clinical differentiation of the two is often difficult if a previous history of HBV is not available. Quantitative measurements...... of immunoglobulin M (IgM) anti-hepatitis B core antibody (anti-HBc) titers and of HBV viral loads (VLs) might allow the separation of AHBV-ALF from CHBV-ALF. Of 1,602 patients with ALF, 60 met clinical criteria for AHBV-ALF and 27 for CHBV-ALF. Sera were available on 47 and 23 patients, respectively. A quantitative...... immunoassay was used to determine IgM anti-HBc levels, and real-time polymerase chain reaction (rtPCR) was used to determine HBV VLs. AHBV-ALFs had much higher IgM anti-HBc titers than CHBV-ALFs (signal-to-noise [S/N] ratio median: 88.5; range, 0-1,120 versus 1.3, 0-750; P

  18. IgM, IgA, IgG1 and IgG2 specific responses in blood and gut secretion of calves fed soyabean products.

    Science.gov (United States)

    Dréau, D; Lallès, J P; Salmon, H; Toullec, R

    1995-07-01

    Calves fed soya proteins may develop severe gastrointestinal disorders. Whether these are predominantly associated with particular Ig subclasses and (or) dietary proteins remains unclear. Therefore, antibody responses to soyabean protein were analysed by dot- and blot-immunobinding in plasma and intestinal mucous secretions. One-month-old calves were fed for 2.5 months liquid diets based on skim milk powder (SMP) or a mixture (2:3, protein basis) of whey and soyabean products including a low antigenic hydrolysed soya protein isolate (HSPI) and a highly antigenic heated soya flour (HSF). Specific antibodies (Abs) of the main isotypes (IgM, IgA, IgG1, IgG2) were characterised by immunostaining of samples which had been previously incubated with nitrocellulose sheets coated with SMP, HSPI or HSF extracts. Plasma collected before feeding experimental diets showed very little specific Abs. By contrast, 2.5 months later, a three-fold increase (P calves fed HSF compared with those fed the control or HSPI diet. IgG1 immunoblotting revealed many protein bands from soya in the molecular range of 22-32 and 38-42 kDa. Immunorecognition of specific proteins from SMP and HSPI remained low and similar among animal groups. Specific IgM, IgA and IgG1 titres against HSF, and to a lesser extent HSPI, were significantly higher (P calves fed HSF compared with other groups. Secretions from calves fed HSF bound to many soyabean proteins in the range of 17-23 and 26-38 kDa, with similar patterns for IgA and IgG1. By contrast, only weak bands were found for IgM and IgG2 in all groups of calves. Thus, calves fed antigenic HSF do present specific Abs including IgG1 and IgA isotypes, both systemically and locally. Therefore, IgG1 and (or) IgA rather than IgM and IgG2 Abs may be preferred for assessing the immunogenicity of soyabean products in calves. Interestingly, soyabean immunogenicity was drastically reduced by adequate proteolysis.

  19. Quantitative immunohistochemical assessment of IgA, IgM, IgG and antigen-specific immunoglobulin secreting plasma cells in pig small intestinal lamina propria.

    Science.gov (United States)

    Bianco, C; Felice, V; Panarese, S; Marrocco, R; Ostanello, F; Brunetti, B; Muscatello, L V; Leotti, G; Vila, T; Joisel, F; Sarli, G

    2014-08-15

    Intestinal immune response plays an important defensive role for pathogens, particularly for those transmitted by the oro-faecal route or for foecal shedding modulation. This work examined three parts of intestine from twelve gilts experimentally infected with PCV2-spiked semen, six vaccinated (V group) and six unvaccinated (NV group) against PCV2, 29 and 53 days post infection (DPI). An immunohistochemical investigation for IgA-, IgG- and IgM-antibody bearing plasma cells (PCs) was run on intestinal samples coupled with a sandwich immunohistochemical method to reveal anti-PCV2 antibody-secreting PCs. Plasma cell density was compared in the two groups of animals at 29 and 53 DPI. The IgA, IgG and IgM PC density did not differ between groups but displayed an increase from the upper (villus) to the lower part of the crypts while a decreasing trend in PC density was identified from duodenum to ileum. In the NV group, no increase in anti-PCV2 PC density was demonstrable in the two sampling moment: the amounts of lamina propria PCV2-specific antibody-producing PCs remained constant, 10.55 ± 4.24 and 10.06 ± 5.01 at 29 DPI and 53 DPI, respectively. In the V group a significant increase in PCV2-specific antibody-producing PCs was observed over time. The amounts of PCV2-specific antibody-producing PCs increased from 9.37 ± 13.36 at 29 DPI to 18.76 ± 15.83 at 53 DPI. The data on IgA, IgM and IgG PC counts can be considered reference values in a population of adult pigs. The sandwich method can be proposed as a technique able to identify specific antibody-secreting PCs in formalin-fixed paraffin-embedded tissues. A practical application of the sandwich method is the demonstration of a "booster-like" response of the lamina propria in vaccinated compared to unvaccinated animals. After virus challenge, vaccination induced an increase in the number of PCs containing specific anti-PCV2 antibodies at the level of intestinal mucosa. Copyright © 2014 Elsevier B.V. All rights

  20. Comparison of R5 and G12 Antibody-Based ELISA Used for the Determination of the Gluten Content in Official Food Samples

    Directory of Open Access Journals (Sweden)

    Rupert Hochegger

    2015-11-01

    Full Text Available Celiac Disease (CD is one of the most common food intolerances. It comes along with serious damage of the mucosa in the small intestine and is caused by the storage proteins—termed “gluten”—of wheat, rye, barley and possibly oats. Sensitive individuals need to stick to a strict gluten-free diet. The gluten level in food products labeled as “gluten-free”, must not exceed 20 mg/kg. It is obvious that effective test methods are needed to accurately determine the gluten concentration in foods. The determination of the presence of gluten in foodstuffs is mainly done by means of an immunochemical method called ELISA (enzyme-linked immunosorbent assay. To check the suitability of a G12 antibody-based gluten detection kit for its use in official control systems a number of routine samples were tested in parallel with two different test kits, as would be done in a routine lab. The determination of the gluten content was performed on samples entering the official laboratory including samples from official control plans, commercially available and private samples to request gluten-free labels. The results obtained with the G12 antibody ELISA assay were comparable to the official R5 method. A validation of the two different methods was not part of this study.

  1. Detection of parvovirus B19 IgM in patients with sickle cell disease in Lagos, Nigeria

    Directory of Open Access Journals (Sweden)

    Christianah Idowu Ayolabi

    2017-07-01

    Full Text Available Objective: To determine anti-B19V IgM seroprevalence among sickle cell disease (SCD patients and patients with other blood genotypes. Methods: In this study, a commercially available ELISA kit (Parvovirus B19 RIDASCREEN Biopharma, Germany was used to analyze 93 serum samples, comprising 68 SCD patients and 25 control samples from individuals with other blood genotypes aged between 1 and 35 years. Results: Anti-B19V IgM was detected in 10 (~15% samples of the SCD patients, in which amongst the SC, 1 (25% positive was recorded, CC had 2 (~67% positive cases and SS had 7 (~11% positives. Among the study control group, a 16% seropositivity was observed, in which among those with genotype AA, 1 (20% was positive, for those with AS, 1 (~7% positive was recorded, for AC, 3 (50% were positive. The age range of 11–20 years showed a positivity of 3 (~4%, those of 21–30 years showed 12 (~24% positivity and both 1–10 years and > 30 years had no positive case recorded. Conclusions: This study showed that B19V is a significant blood borne pathogen in our environment and concerted effort is needed to advance strategies to curb this infection and diminish the common severe complications usually associated with this virus infection among SCD patients.

  2. A small antigenic determinant of the Chikungunya virus E2 protein is sufficient to induce neutralizing antibodies which are partially protective in mice.

    Directory of Open Access Journals (Sweden)

    Christopher Weber

    2015-04-01

    Full Text Available The mosquito-borne Chikungunya virus (CHIKV causes high fever and severe joint pain in humans. It is expected to spread in the future to Europe and has recently reached the USA due to globalization, climate change and vector switch. Despite this, little is known about the virus life cycle and, so far, there is no specific treatment or vaccination against Chikungunya infections. We aimed here to identify small antigenic determinants of the CHIKV E2 protein able to induce neutralizing immune responses.E2 enables attachment of the virus to target cells and a humoral immune response against E2 should protect from CHIKV infections. Seven recombinant proteins derived from E2 and consisting of linear and/or structural antigens were created, and were expressed in and purified from E. coli. BALB/c mice were vaccinated with these recombinant proteins and the mouse sera were screened for neutralizing antibodies. Whereas a linear N-terminally exposed peptide (L and surface-exposed parts of the E2 domain A (sA alone did not induce neutralizing antibodies, a construct containing domain B and a part of the β-ribbon (called B+ was sufficient to induce neutralizing antibodies. Furthermore, domain sA fused to B+ (sAB+ induced the highest amount of neutralizing antibodies. Therefore, the construct sAB+ was used to generate a recombinant modified vaccinia virus Ankara (MVA, MVA-CHIKV-sAB+. Mice were vaccinated with MVA-CHIKV-sAB+ and/or the recombinant protein sAB+ and were subsequently challenged with wild-type CHIKV. Whereas four vaccinations with MVA-CHIKV-sAB+ were not sufficient to protect mice from a CHIKV infection, protein vaccination with sAB+ markedly reduced the viral titers of vaccinated mice.The recombinant protein sAB+ contains important structural antigens for a neutralizing antibody response in mice and its formulation with appropriate adjuvants might lead to a future CHIKV vaccine.

  3. Malaria serology: performance of six Plasmodium falciparum antigen extracts and of three ways of determining serum titers in IgG and IgM-ELISA Sorologia da malária: desempenho de seis diferentes extratos antigênicos de Plasmodium falciparum e de três métodos de determinação de títulos de soros no teste ELISA-IgG e IgM

    Directory of Open Access Journals (Sweden)

    Maria Carmen Arroyo Sanchez

    1993-12-01

    Full Text Available The study evaluated six Plasmodium falciparum antigen extracts to be used in the IgG and IgM enzyme-linked immunosorbent assays (ELISA, for malaria diagnosis and epidemiological studies. Results obtained with eighteen positive and nine negative control sera indicated that there were statistically significant differences among these antigen extracts (Multifactor ANOVA, pEste estudo avaliou seis extratos antigênicos de Plasmodium falciparum para utilização no diagnóstico da malária e em estudos epidemiológicos, empregando o teste "enzyme-linked immunosorbent assay" (ELISA para a pesquisa de anticorpos das classes IgG e IgM. Os resultados obtidos empregando dezoito soros padrão positivo e nove padrão negativo, indicaram que houve diferenças estatisticamente significativas entre os extratos antigênicos estudados (Multifactor ANOVA, p <0,0001. Extratos antigênicos preparados com uréia, desoxicolato de sódio e Zwittergent forneceram melhores resultados que os demais e se comportaram de modo bastante semelhante, para a detecção de anticorpos IgG. Extratos antigenicos de uréia, alcalino e de desoxicolato de sódio foram os melhores na detecção de anticorpos IgM. Devido à relação linear obtida entre as densidades ópticas (ou os log10 respectivos e os log10 das diluições, com uma inclinação constante, os títulos dos soros foram determinados através de duas equações diferentes, que necessitam apenas de uma diluição do soro. Para anticorpos IgM, a expressão que emprega log10 da densidade óptica forneceu resultados que se correlacionaram melhor com a titulação (95% Bon-ferroni. Para anticorpos IgG, as diferenças entre resultados não foram significativas. A avaliação da reprodutibilidade dos títulos e das partidas de antígenos forneceu resultados satisfatórios.

  4. Immune inhibition of virus release from human and nonhuman cells by antibody to viral and host cell determinants.

    Science.gov (United States)

    Shariff, D M; Davies, J; Desperbasques, M; Billstrom, M; Geerligs, H J; Welling, G W; Welling-Wester, S; Buchan, A; Skinner, G R

    1991-01-01

    Immune inhibition of release of the DNA viruses, herpes simplex virus types 1 and 2 and pseudorabies virus by anti-viral and anti-host cell sera occurred while two RNA viruses, influenza and encephalomyocarditis, were inhibited only by anti-viral sera (not anti-host cell sera). Simian virus 40 and surprisingly two herpes viruses, bovine mamillitis and equine abortion, were not inhibited by either anti-viral or anti-host sera. Using the herpes simplex virus model, inhibition of virus release was detected in different cells of human and nonhuman origin with cross-inhibition between cell lines of different origin; thus, this form of immunotherapy may not require antibody to be tissue or organ specific. Evidence of inhibition of virus release from neoplastic and leukemic cell lines suggests possible application of this approach to control of virus-mediated leukoproliferative pathology (e.g. Burkitt's lymphoma or adult T cell leukemia).

  5. Pathogenesis and mechanisms of antibody-mediated hemolysis.

    Science.gov (United States)

    Flegel, Willy A

    2015-07-01

    The clinical consequences of antibodies to red blood cells (RBCs) have been studied for a century. Most clinically relevant antibodies can be detected by sensitive in vitro assays. Several mechanisms of antibody-mediated hemolysis are well understood. Such hemolysis after transfusion is reliably avoided in a donor-recipient pair, if one individual is negative for the cognate antigen to which the other has the antibody. Mechanisms of antibody-mediated hemolysis were reviewed based on a presentation at the Strategies to Address Hemolytic Complications of Immune Globulin Infusions Workshop addressing intravenous immunoglobulin (IVIG) and ABO antibodies. The presented topics included the rates of intravascular and extravascular hemolysis; immunoglobulin (Ig)M and IgG isoagglutinins; auto- and alloantibodies; antibody specificity; A, B, A,B, and A1 antigens; A1 versus A2 phenotypes; monocytes-macrophages, other immune cells, and complement; monocyte monolayer assay; antibody-dependent cell-mediated cytotoxicity; and transfusion reactions due to ABO and other antibodies. Several clinically relevant questions remained unresolved, and diagnostic tools were lacking to routinely and reliably predict the clinical consequences of RBC antibodies. Most hemolytic transfusion reactions associated with IVIG were due to ABO antibodies. Reducing the titers of such antibodies in IVIG may lower the frequency of this kind of adverse event. The only way to stop these events is to have no anti-A or anti-B in the IVIG products. © 2015 AABB.

  6. Theobromine Is Responsible for the Effects of Cocoa on the Antibody Immune Status of Rats.

    Science.gov (United States)

    Camps-Bossacoma, Mariona; Pérez-Cano, Francisco J; Franch, Àngels; Castell, Margarida

    2018-03-01

    A 10% cocoa-enriched diet influences immune system functionality including the prevention of the antibody response and the induction of lower immunoglobulin (Ig) concentrations. However, neither cocoa polyphenols nor cocoa fiber can totally explain these immunoregulatory properties. This study aimed to establish the influence of cocoa theobromine in systemic and intestinal Ig concentrations and to determine the effect of cocoa or theobromine feeding on lymphoid tissue lymphocyte composition. Three-week-old female Lewis rats were fed either a standard diet (AIN-93M; RF group), a 10% cocoa diet (CC group), or a 0.25% theobromine diet (the same amount provided by the cocoa diet; TB group) in 2 separate experiments that lasted 19 (experiment 1) or 8 (experiment 2) d. Serum IgG, IgM, IgA, and intestinal secretory IgA (sIgA) concentrations were determined. In addition, at the end of experiment 2, thymus, mesenteric lymph node (MLN), and spleen lymphocyte populations were analyzed. Both CC and TB groups in experiments 1 and 2 showed similar serum IgG, IgM, and IgA and intestinal sIgA concentrations, which were lower than those in the RF group (46-98% lower in experiment 1 and 23-91% lower in experiment 2; P theobromine diets similarly changed the thymocyte composition by increasing CD4-CD8- (+133%) and CD4+CD8- (+53%) proportions (P theobromine in cocoa plays an immunoregulatory role that is responsible for cocoa's influence on both systemic and intestinal antibody concentrations and also for modifying lymphoid tissue lymphocyte composition in young healthy Lewis rats. The majority of these changes are observed after a single week of being fed a diet containing 0.25% theobromine.

  7. An amperometric immunosensor for diagnosis of celiac disease based on covalent immobilization of open conformation tissue transglutaminase for determination of anti-tTG antibodies in human serum.

    Science.gov (United States)

    Giannetto, Marco; Mattarozzi, Monica; Umiltà, Eleonora; Manfredi, Anita; Quaglia, Sara; Careri, Maria

    2014-12-15

    A new amperometric immunosensor based on the covalent immobilization of tissue transglutaminase enzyme in its open conformation (open-tTG) was developed and optimized for determination of anti-tissue transglutaminase antibodies (anti-tTG) in human serum. Experimental design allowed us to find the optimal conditions for quantification of both IgA and IgG isotypes of anti-tTG in order to assess suitability of the device for diagnostic purposes. The glassy carbon electrodic substrate was electrochemically functionalized with gold nanoparticles and subsequently derivatized with a self-assembled monolayer of 11-mercaptoundecanoic acid for the covalent anchoring of the enzyme. This step was performed under carefully controlled conditions in order to keep the open conformation of the tTG. The immunosensor showed good analytical performance with limit of detection levels (1.7 AU mL(-1) for IgA and 2.7 AU mL(-1) for IgG) below the diagnostic threshold value (3.0 AU mL(-1)) and inter-sensor reproducibility giving RSD lower than 10%. The developed sensor was validated in serum samples from pediatric patients for clinical applications, using two ELISA kits specific for the determination of anti-tTG IgA and IgG antibodies as reference methods; good recovery rates ranging from 74% to 117% were calculated. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Role of antilipopolysaccharide antibodies in serum bactericidal activity against Salmonella enterica serovar Typhimurium in healthy adults and children in the United States.

    Science.gov (United States)

    Trebicka, Estela; Jacob, Susan; Pirzai, Waheed; Hurley, Bryan P; Cherayil, Bobby J

    2013-10-01

    Recent observations from Africa have rekindled interest in the role of serum bactericidal antibodies in protecting against systemic infection with Salmonella enterica serovar Typhimurium. To determine whether the findings are applicable to other populations, we analyzed serum samples collected from healthy individuals in the United States. We found that all but 1 of the 49 adult samples tested had robust bactericidal activity against S. Typhimurium in a standard in vitro assay. The activity was dependent on complement and could be reproduced by immunoglobulin G (IgG) purified from the sera. The bactericidal activity was inhibited by competition with soluble lipopolysaccharide (LPS) from S. Typhimurium but not from Escherichia coli, consistent with recognition of a determinant in the O-antigen polysaccharide. Sera from healthy children aged 10 to 48 months also had bactericidal activity, although it was significantly less than in the adults, correlating with lower levels of LPS-specific IgM and IgG. The lone sample in our collection that lacked bactericidal activity was able to inhibit killing of S. Typhimurium by the other sera. The inhibition correlated with the presence of an LPS-specific IgM and was associated with decreased complement deposition on the bacterial surface. Our results indicate that healthy individuals can have circulating antibodies to LPS that either mediate or inhibit killing of S. Typhimurium. The findings contrast with the observations from Africa, which linked bactericidal activity to antibodies against an S. Typhimurium outer membrane protein and correlated the presence of inhibitory anti-LPS antibodies with human immunodeficiency virus infection.

  9. Effect of the Various Steps in the Processing of Human Milk in the Concentrations of IgA, IgM, and Lactoferrin.

    Science.gov (United States)

    Arroyo, Gerardo; Ortiz Barrientos, Kevin Alexander; Lange, Karla; Nave, Federico; Miss Mas, Gabriela; Lam Aguilar, Pamela; Soto Galindo, Miguel Angel

    2017-09-01

    Human milk immune components are unique and important for the development of the newborn. Milk processing at the Human Milk Banks (HMB), however, causes partial destruction of immune proteins. The objective of this study was to determine the effects that heating during the milk processing procedure at the HMB had on the concentrations of IgA, IgM, and lactoferrin at three critical points in time. Fifty milk samples (150 mL) were collected from voluntary donors at the HMB at the Hospital Nacional Pedro de Bethancourt, located in Antigua Guatemala. Samples from three critical points in time during the milk processing procedure were selected for analysis: freezing/thawing I, freezing/thawing II, and pasteurization. IgA, IgM, and lactoferrin concentrations were determined during each critical point and compared with a baseline concentration. After milk processing, IgA, IgM, and lactoferrin mean concentrations were reduced by 30.0%, 36.0%, and 70.0%, respectively (p milk processing on the immune proteins that were evaluated in this study demonstrated a significant reduction.

  10. Caracterización de IgM, IgG total, IgG1 y anticuerpos de cadena pesada en calostro de llamas (Lama Glama) mediante Elisa

    OpenAIRE

    Caggiano, Nicolás; Saccodossi, Natalia; Gentile, Maria Teresa; Chiappe Barbará, María Angelina; Leoni, Juliana; de Simone, Emilio Adrian

    2016-01-01

    Objetivos: determinar los niveles de IgM, IgG total y Anticuerpos de Cadena Pesada (HCAbs; por su sigla en inglés Heavy Chain Antibodies) (IgG2 e IgG3) en calostro de llamas y evaluar la concentración de HCAbs en relación a la IgG total y al isotipo convencional IgG1. Métodos: en este estudio se utilizaron 15 llamas preñadas, que fueron ordeñadas dentro de las primeras 24 horas post-parto. Se diseñaron ELISAs Sandwich para la cuantificación de IgM total, IgG total e IgG1. La concent...

  11. Poroid Fungi of Hungary in the Collection of Zoltán Igmándy

    Directory of Open Access Journals (Sweden)

    SZABÓ, Ilona

    2012-01-01

    Full Text Available Zoltán Igmándy (1925–2000, prominent Hungarian mycologist, worked as professor of forest protection at the University of West-Hungary Sopron. His main research area was the investigation of wood-inhabiting poroid fungi of Hungary, their occurrence and importance in forest pathology and wood protection. During his 40 years of scientific activity Igmándy created a rich fungal collection (herbarium Z. Igmándy which includes the complete polypore mycota known to occur in Hungary until 1990. The paper provides a brief compendium of the collection, a list of species characterised by the number of specimens, number of sampling locations and enumeration of the hosts and substrata of the specimens.

  12. Symptomatic Secondary Selective IgM Immunodeficiency in Adult Man with Undiagnosed Celiac Disease

    Directory of Open Access Journals (Sweden)

    Eli Magen

    2012-01-01

    Full Text Available Selective IgM immunodeficiency (SIgMID is a heterogeneous disorder with no known genetic background and may occur as a primary or a secondary condition. Celiac disease has been reported in association with several humeral immunodeficiencies, including isolated severe selective IgA deficiency, panhypogammaglobulinemia, and isolated combined IgA and IgM deficiency. There are only few reported cases of pediatric and adult patients with SIgMID and celiac disease. In this paper, we describe an adult patient with a symptomatic secondary SIgMID associated with undiagnosed celiac disease, with a resolution of clinical symptoms of immunodeficiency and serum IgM normalization following a gluten-free diet.

  13. Nitrocellulose membrane-based enzyme-linked immunoassay for dengue serotype-1 IgM detection

    International Nuclear Information System (INIS)

    Leon, S.; Guevara, C.; Chunga, A.

    1999-01-01

    To evaluate the sensitivity and specifity of a nitrocellulose membrane-based immunoassay for dengue IgM, with respect to capture enzyme immunoassay, for the diagnosis of dengue virus infection. 101 serum samples were processed and divided into 2 groups: 53 from dengue serotype 1 (DEN1) infected patients, and 48 from healthy subjects. Both groups were tested with a nitrocellulose membrane-based IgM capture enzyme immunoassay (NMB-EIA) and also with an ELISA as referential pattern. NMB-EIA testing detected IgM anti-DEN1 in 94,34% of samples from infected patients, and in 14,58% of control samples, whereas ELISA fails to report false positive or false negative results: NMB-EIA appears to be a good alternative for dengue infection diagnosis. (authors)

  14. Prevalence and isotype distribution of antiphospholipid antibodies in unselected Chilean patients with venous and arterial thrombosis.

    Science.gov (United States)

    Palomo, Iván; Pereira, Jaime; Alarcón, Marcelo; Vásquez, Marcela; Pinochet, Carmen; Vélez, María T; Sandoval, Jorge; Icaza, Gloria; Pierangeli, Silvia

    2004-04-01

    Antiphospholipid antibodies (aPL) are a heterogeneous family of antibodies associated with thrombotic events and other complications. The objective of this study was to investigate the prevalence of aPL in a group of Chilean patients with thrombosis. Two hundred and twenty-six patients with venous and arterial thrombosis and 95 healthy controls were studied. Anticardiolipin (aCL), anti-beta(2 )glycoprotein I (anti-beta(2)GPI), and antiprothrombin (aPT) antibodies were determined. Eighty-eight out of 226 (38.9%) patients with thrombosis had some type of aPL. Fifty-seven patients (25.2%) were positive for aCL, 31 (13.7%) for aPT, and 14 (6.2%) for anti-beta(2)GPI antibodies. Twelve patients (5.3%) were positive for more than one aPL. IgG, IgM and IgA isotypes were observed in aCL, anti-beta(2)GPI, and aPT antibodies. Twenty-six out of 92 (28.3%) patients with venous thrombosis and 31/134 (23.1%) patients with arterial thrombosis were positive for aCL antibodies. With regard to the control group (4/95=4.2%), the odd ratios (OR) were 5.2 (1.3-19.8; p0.01) and 5.7 (1.6-22.3; p0.01), respectively. Additionally, we observed statistically significant OR with aPT and anti-beta(2)GPI antibodies; in the first, with venous and arterial thrombosis, and in the second, only with arterial thrombosis. Our results show a significant prevalence of aPL, predominantly aCL and aPT antibodies, in patients with thrombosis. Additionally, aCL and aPT antibodies appear to be a risk factor for venous and arterial thrombosis, and anti-beta(2)GPI antibodies appear to be a risk factor for arterial thrombosis.

  15. Increased urine IgM excretion predicts cardiovascular events in patients with type 1 diabetes nephropathy

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    Torffvit Ole

    2009-08-01

    Full Text Available Abstract Background Diabetic nephropathy, a major complication of diabetes, is characterized by progressive renal injury and increased cardiovascular mortality. An increased urinary albumin excretion due dysfunction of the glomerular barrier is an early sign of diabetic nephropathy. An increased urinary excretion of higher molecular weight proteins such as IgM appears with progression of glomerular injury. We aim here to study the prognostic significance of urine IgM excretion in patients with type 1 diabetes mellitus (type 1 diabetic nephropathy. Methods This is an observational study of 139 patients with type1 diabetes mellitus (79 males and 60 females under routine care at the diabetic outpatient clinic at the Lund University Hospital. The median follow-up time was 18 years (1 to 22 years. Urine albumin and urine IgM concentration were measured at time of recruitment. Results Overall 32 (14 male and 18 female patients died in a cardiovascular event and 20 (11 male and 9 female patients reached end-stage renal disease. Univariate analysis indicated that patient survival and renal survival were inversely associated with urine albumin excretion (RR = 2.9 and 5.8, respectively and urine IgM excretion (RR = 4.6 and 5.7, respectively. Stratified analysis demonstrated that in patients with different degrees of albuminuria, the cardiovascular mortality rate and the incidence of end-stage renal disease was approximately three times higher in patients with increased urine IgM excretion. Conclusion An increase in urinary IgM excretion in patients with type 1 diabetes is associated with an increased risk for cardiovascular mortality and renal failure, regardless of the degree of albuminuria.

  16. Helium Reionization Simulations. II. Signatures of Quasar Activity on the IGM

    Science.gov (United States)

    La Plante, Paul; Trac, Hy; Croft, Rupert; Cen, Renyue

    2017-06-01

    We have run a new suite of simulations that solve hydrodynamics and radiative transfer simultaneously to study helium II reionization. Our suite of simulations employs various models for populating quasars inside of dark matter halos, which affect the He II reionization history. In particular, we are able to explore the impact that differences in the timing and duration of reionization have on observables. We examine the thermal signature that reionization leaves on the intergalactic medium (IGM), and measure the temperature-density relation. As previous studies have shown, we confirm that the photoheating feedback from helium II reionization raises the temperature of the IGM by several thousand kelvin. To compare against observations, we generate synthetic Lyα forest sightlines on-the-fly and match the observed effective optical depth {τ }{eff}(z) of hydrogen to recent observations. We show that when the simulations have been normalized to have the same values of {τ }{eff}, the effect that helium II reionization has on observations of the hydrogen Lyα forest is minimal. Specifically, the flux PDF and the one-dimensional power spectrum are sensitive to the thermal state of the IGM, but do not show direct evidence for the ionization state of helium. We show that the peak temperature of the IGM typically corresponds to the time of 90%-95% helium ionization by volume, and is a relatively robust indicator of the timing of reionization. Future observations of helium reionization from the hydrogen Lyα forest should thus focus on measuring the temperature of the IGM, especially at mean density. Detecting the peak in the IGM temperature would provide valuable information about the timing of the end of helium II reionization.

  17. 18-FDG-PET in pretherapeutical determination of extra medullary involvement prior to Re-188-antibody guided myeloablative therapy

    International Nuclear Information System (INIS)

    Hofmann, M.; Boerner, A.R.; Otto, D.; Knapp, W.H.; Hertenstein, B.

    2002-01-01

    Full text: In relapsed or refractory leukemia it is highly desirable to intensify the conditioning prior to allogenic bone mamarrow transplantation (BMT). Application of the 188-Re-labelled antibody BW 250/183 has proved to be feasible for internal radiation therapy of the bone marrow. Because the number of granulocytes (which express the CD66b antigen targeted) in extramedullary manifestations are usually low, the treatment of extramkedullary lesions will be not effective. This study deals with the pretherapeutical identification of patients with extramedullary involvement via 18-F-FDG-PET. 1-2 weeks prior scheduled 188-Re-therapy 12 patients underwent 18-F-FDG PET (dedicated PET scanner Siemens ECAT EXACT 47). Non physiological tracer accumulations were evaluated by means of SUV and consecutive morphological imaging (CT/MRI). 3 of 11 patients did show non physiological enrichment. 1 patient had enrichment in the upper mediastine, which was confirmed as extramedullary leukemia involvement by cytology. In CT scan these lesions had been identified as small lymphnodes (1-2 cm diameter). 2 Patients did show small lesions in the lung. In one patient they were confirmed to be active fungous infection (pos. sputum culture) and in the other patient they were regarded as regenerative residuals of previous known pneumonic infection not active now (neg. sputum cultures and neg. inflammation markers). 18-F-FDG PET is helpful of identifying extramedullary involvement and gives in addition information on the inflammatory status of patients prior to BMT. (author)

  18. Renal disease, epidermal necrosis, and epithelial cell antibodies.

    OpenAIRE

    Deal, J E; Groves, R W; Harmer, A W; Welsh, K I; MacDonald, D M; Rigden, S P

    1991-01-01

    OBJECTIVE--To describe the association between epithelial cell IgM, which has previously been associated with an increased incidence of loss of renal graft in children, with a novel cutaneous eruption and unexplained native renal disease. DESIGN--Observational study on children with epithelial cell antibody presenting with unexplained renal or skin disease. SETTING--General paediatric department and regional paediatric nephrology unit. PATIENTS--Six children (five girls, one boy), who present...

  19. Genetic differences in natural antibody levels in common carp (Cyprinus carpio L.)

    NARCIS (Netherlands)

    Kachamakova, N.M.; Irnazarow, I.; Parmentier, H.K.; Savelkoul, H.F.J.; Pilarczyk, A.; Wiegertjes, G.F.

    2006-01-01

    In mammals, natural antibodies (Nabs) are mostly of the IgM isotype and can bind to a particular antigen or pathogen even if the host has never been exposed. Despite their early detection and abundance, the exact role and genetic control of Nabs remain unclear. We have used an indirect ELISA with

  20. Genetic parameters for natural antibody isotype titers in milk of Dutch Holstein-Friesians

    NARCIS (Netherlands)

    Wijga, S.; Bovenhuis, H.; Bastiaansen, J.W.M.; Arendonk, van J.A.M.; Ploegaert, T.C.W.; Tijhaar, E.; Poel, van der J.J.

    2013-01-01

    The objective of the present study was to estimate genetic parameters for natural antibody isotypes immunoglobulin (Ig) A, IgG1 and IgM titers binding the bacterial antigens lipopolysaccharide, peptidoglycan and the model antigen keyhole limpet hemocyanin in Dutch Holstein-Friesian cows (n = 1695).

  1. Comparison of dot-ELISA and standard ELISA for detection of Neisseria meningitidis outer membrane complex-specific antibodies

    Directory of Open Access Journals (Sweden)

    Elza FT Belo

    Full Text Available Dot-ELISA using the outer membrane complex antigens of Neisseria meningitidis as a target was standardized for rapid detection of meningococcal-specific antibodies in human serum. We investigated the level of meningococcal-specific IgG, IgA, and IgM in serum using dot-ELISA with outer membrane antigens prepared from Neisseria meningitidis serotype B:4.19:P1.15,3,7,9 (a strain isolated from a Brazilian epidemic. The dot-ELISA is based on the same principles as the standard ELISA and is useful for detection of anti-N. meningitidis B antibodies in serum of patients with meningococcal infections. For the assay, outer membrane complexes (OMCs were absorbed by nitrocellulose membrane and blocked with a 5% skim milk solution. Serum samples were drawn upon hospital admission and during convalescence from patients with meningococcal septicemia, and single samples were drawn from uninfected controls. We retrospectively examined a total of 57 serum samples: 35 from patients infected with N. meningitidis B, 12 from patients infected with Haemophilus influenzae b, and 10 from health individuals. When performed at room temperature, dot-ELISA took approximately four hours to perform, and the optimum antigen concentration was 0.42 µg per dot. The specificity of IgG, IgM, and IgA demonstrates that dot-ELISA using OMCs from N. meningitidis B as a target is suitable for serologic verification of clinically suspected meningococcal disease in patients and for titer determination of antibodies produced during different phases of natural infection. Furthermore, the sensitivity of dot-ELISA was comparable to that of standard ELISA. Overall, dot-ELISA is simple to perform, rapid, and low cost. Further validation of the test as a screening tool is required.

  2. A single serum dilution enzyme-linked immunosorbent assay for determining anti-human papillomavirus (HPV) antibody titres in humans immunised with prophylactic HPV vaccines.

    Science.gov (United States)

    Jin, Yingji; Kim, Hyoung Jin; Yim, Ga Won; Kim, Young Tae; Chang, Don Yong; Kim, Hong-Jin

    2012-07-01

    Two types of prophylactic human papillomavirus (HPV) vaccines are currently available. However, there is no simple monitoring system for assessing acquired immunity that can cope simultaneously with large numbers of serum samples. Approximately 30% of women with normal cytology are known to be seropositive for HPV types 16 and 18 because of the high prevalence of these HPV types. Therefore, to be useful the monitoring system has to discriminate clearly between vaccine recipients and other serology groups. However, there has never been any focus on developing a method to satisfy this condition. In this study, we developed a high-throughput single-serum-dilution enzyme-linked immunoassay (ELISA) system for determining anti-HPV antibody titres following vaccination. We optimised the conditions for each ELISA step to increase its accuracy and precision and to avoid the high background of non-specific reactions that is a major problem for serology assays. The new ELISA system has superior linearity, accuracy and reproducibility. Moreover, it clearly discriminated between antibody levels in vaccine recipients and those in other serology groups such as individuals with normal cervical cytology and those with cervical cancer. Therefore, this single-serum-dilution ELISA should be very useful for assessing the acquired immunity of HPV vaccine recipients. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. Thyroglobulin (Tg) recovery testing with quantitative Tg antibody measurement for determining interference in serum Tg assays in differentiated thyroid carcinoma.

    Science.gov (United States)

    Persoon, Adrienne C M; Links, Thera P; Wilde, Juergen; Sluiter, Wim J; Wolffenbuttel, Bruce H R; van den Ouweland, Johannes M W

    2006-06-01

    Thyroglobulin (Tg) measurements are complicated by interference from Tg autoantibodies (TgAbs) or heterophilic antibodies (HAMAs). We used a new automated immunochemiluminometric assay (ICMA) with Tg recovery (TgR) on the Nichols Advantage platform to reassess the clinical utility of recovery testing in detecting interference in serum Tg measurement in patients with differentiated thyroid carcinoma. We used 2 TgAb methods to detect Tg measurement interference with TgR and quantitative TgAb measurement in sera from 127 patients. In a limited number of samples, we used an RIA as comparison method because it appeared to be minimally affected by TgAb. Prevalence of TgAbs was 13% (17 of 127) in either 1 or both TgAb assays. A compromised TgR ( or =70%) corresponded with TgAb negativity in both assays for 95 of 101 samples (94%). In 6 TgAb-positive sera with TgR within the reference interval, there were no discrepancies between RIA and ICMA results. We obtained discordant RIA and ICMA results for 6 of 9 TgAb-positive sera with decreased TgR. In 1 TgAb-negative sample, the Tg result was falsely increased because of interference by HAMAs, as shown by an overrecovery of 126%. The Nichols Advantage TgR assay is a valuable complementary method to overcome the technical problem of interference by TgAbs or HAMAs in TgAb assays. Further studies are needed to confirm the potential added value of this TgR assay.

  4. Magnitude of Increased Infliximab Clearance Imposed by Anti-infliximab Antibodies in Crohn's Disease Is Determined by Their Concentration.

    Science.gov (United States)

    Edlund, Helena; Steenholdt, Casper; Ainsworth, Mark A; Goebgen, Eva; Brynskov, Jørn; Thomsen, Ole Ø; Huisinga, Wilhelm; Kloft, Charlotte

    2017-01-01

    Antibodies (Abs) against infliximab (IFX) increase IFX clearance and can result in treatment failure and acute hypersensitivity reactions. However, interpretation of their clinical value is complicated by individual differences in Ab responses and methods used for quantification. The increase in IFX clearance imposed by anti-IFX Abs has generally been evaluated using a binary classification, i.e., positive or negative. This analysis aimed to investigate if anti-IFX Ab concentrations provide a more adequate prediction of alterations in clearance. Data originated from a clinical trial on Crohn's disease patients with IFX treatment failure. The trial was not originally designed for pharmacokinetic analysis. Therefore, published pharmacokinetic models were utilized as priors to enable covariate investigation. The impact of anti-IFX Abs on clearance was assessed using different mathematical relationships and exploiting information from two different quantification assays, measuring semi-quantitative "total" or "unbound neutralizing" concentrations of anti-IFX Ab, respectively. Inclusion of anti-IFX Ab status/concentration improved the model's performance for all investigated relationships. The anti-IFX Ab concentrations were superior to the binary classifications, indicating that the magnitude of increase in IFX clearance imposed by anti-IFX Abs closely relates to their concentration. Furthermore, total anti-IFX Ab concentrations appeared superior to the unbound neutralizing fraction in identifying high clearance individuals. Simulations showed that even at low concentrations, anti-IFX Abs lead to sub-therapeutic IFX concentrations, supporting a need of treatment interventions in all anti-IFX Ab positive patients. The developed model can serve as a basis for further investigations to refine treatment recommendations for patients with anti-IFX Abs.

  5. Evaluation of tests based on the antibody response to keyhole limpet haemocyanin and soluble egg antigen to differentiate acute and chronic human schistosomiasis mansoni

    Directory of Open Access Journals (Sweden)

    Lílian Beck

    2004-08-01

    Full Text Available Specific IgG and IgM responses to soluble egg antigen (SEA and keyhole limpet haemocyanin (KLH were measured by ELISA in patients with acute and chronic schistosomiasis. The tests based upon IgM and IgG antibodies responses to KLH presented the best diagnostic discrimination, and can be used in conjunction with clinical and epidemiological data to the differential diagnosis of acute schistosomiasis.

  6. Behaviour of non-donor specific antibodies during rapid re-synthesis of donor specific HLA antibodies after antibody incompatible renal transplantation.

    Directory of Open Access Journals (Sweden)

    Nithya S Krishnan

    Full Text Available HLA directed antibodies play an important role in acute and chronic allograft rejection. During viral infection of a patient with HLA antibodies, the HLA antibody levels may rise even though there is no new immunization with antigen. However it is not known whether the converse occurs, and whether changes on non-donor specific antibodies are associated with any outcomes following HLA antibody incompatible renal transplantation.55 patients, 31 women and 24 men, who underwent HLAi renal transplant in our center from September 2005 to September 2010 were included in the studies. We analysed the data using two different approaches, based on; i DSA levels and ii rejection episode post transplant. HLA antibody levels were measured during the early post transplant period and corresponding CMV, VZV and Anti-HBs IgG antibody levels and blood group IgG, IgM and IgA antibodies were quantified.Despite a significant DSA antibody rise no significant non-donor specific HLA antibody, viral or blood group antibody rise was found. In rejection episode analyses, multiple logistic regression modelling showed that change in the DSA was significantly associated with rejection (p = 0.002, even when adjusted for other antibody levels. No other antibody levels were predictive of rejection. Increase in DSA from pre treatment to a post transplant peak of 1000 was equivalent to an increased chance of rejection with an odds ratio of 1.47 (1.08, 2.00.In spite of increases or decreases in the DSA levels, there were no changes in the viral or the blood group antibodies in these patients. Thus the DSA rise is specific in contrast to the viral, blood group or third party antibodies post transplantation. Increases in the DSA post transplant in comparison to pre-treatment are strongly associated with occurrence of rejection.

  7. Prevalence of Serum Antibodies to TORCH Infection in and Around Varanasi, Northern India.

    Science.gov (United States)

    Sen, M R; Shukla, B N; Tuhina, Banerjee

    2012-11-01

    The acute infections which are caused by Toxoplasma gondii, Rubella virus, Cytomegalovirus (CMV) and the Herpes Simplex Virus (HSV-2) during pregnancy are often associated with adverse foetal outcomes and reproductive failures. In the Indian context, the exact seroprevalence of these infections is not known due to unavailability of baseline data. The present study was undertaken to determine the serological evidence of the acute TORCH infections in women who were in the first trimesters of their pregnancies in and around Varanasi, north India. This study was carried out in the Sir Sunderlal Hospital, Varanasi and in the Department of Microbiology, Institute of Medical Sciences, BHU, Varanasi, UP, India. The study population involved pregnant women with bad obstetric histories, who were in the first trimester of their pregnancy. Sera were collected from the women with Bon and they were tested for the presence of specific IgM antibodies against the TORCH infections by ELISA. A 95% confidence interval was calculated for the positive cases in each of the TORCH components. The specific IgM antibodies were found to be positive in 74(19.4%) cases for toxoplasmosis, in 126 (30.4%) cases for the Rubella virus, in 130 (34.7%) cases for CMV and in 151 samples (33.5%) for the HSV-2 infections. The study showed a high prevalence of the infections which were caused by the TORCH complex amongst pregnant women with bad obstetric histories. Therefore, all the antenatal cases should be routinely screened for the TORCH infections, for carrying out early interventions to prevent foetal loss.

  8. CD73 expression identifies a subset of IgM+ antigen-experienced cells with memory attributes that is T cell and CD40 signalling dependent.

    Science.gov (United States)

    D'Souza, Lucas; Gupta, Sneh Lata; Bal, Vineeta; Rath, Satyajit; George, Anna

    2017-12-01

    B-cell memory was long characterized as isotype-switched, somatically mutated and germinal centre (GC)-derived. However, it is now clear that the memory pool is a complex mixture that includes unswitched and unmutated cells. Further, expression of CD73, CD80 and CD273 has allowed the categorization of B-cell memory into multiple subsets, with combinatorial expression of the markers increasing with GC progression, isotype-switching and acquisition of somatic mutations. We have extended these findings to determine whether these markers can be used to identify IgM memory phenotypically as arising from T-dependent versus T-independent responses. We report that CD73 expression identifies a subset of antigen-experienced IgM + cells that share attributes of functional B-cell memory. This subset is reduced in the spleens of T-cell-deficient and CD40-deficient mice and in mixed marrow chimeras made with mutant and wild-type marrow, the proportion of CD73 + IgM memory is restored in the T-cell-deficient donor compartment but not in the CD40-deficient donor compartment, indicating that CD40 ligation is involved in its generation. We also report that CD40 signalling supports optimal expression of CD73 on splenic T cells and age-associated B cells (ABCs), but not on other immune cells such as neutrophils, marginal zone B cells, peritoneal cavity B-1 B cells and regulatory T and B cells. Our data indicate that in addition to promoting GC-associated memory generation during B-cell differentiation, CD40-signalling can influence the composition of the unswitched memory B-cell pool. They also raise the possibility that a fraction of ABCs may represent T-cell-dependent IgM memory. © 2017 John Wiley & Sons Ltd.

  9. IgM, lgG and IL-6 profiles in the Trypanosoma brucei brucei monkey model of human African trypanosomiasis.

    Science.gov (United States)

    Waema, Maxwell W; Maina, Naomi W; Ngotho, Maina; Karanja, Simon M; Gachie, Beatrice M; Maranga, Dawn N; Kagira, John M

    2017-04-01

    Human African trypanosomiasis (HAT) patients manifest immunological profiles, whose variations over time can be used to indicate disease progression. However, monitoring of these biomarkers in human patients is beset by several limitations which can be offset by using chronic animal models. A recent improved monkey model of HAT using a Trypanosoma brucei brucei isolate has been developed but the immunological profile has not been elucidated. The objectives of the current study was to determine the IgM, IgG and IL-6 profiles in blood and cerebrospinal fluid (CSF) in vervet monkeys infected with T. b. brucei. Three vervet monkeys were infected intravenously with 10 5 T. b. brucei, monitored for disease development and subsequently treated 28days post infection (dpi) sub-curatively using diminazene aceturate (DA) to induce late stage disease and curatively treated with melarsoprol (Mel B) at 119 dpi, respectively. Matched serum and cerebrospinal fluid (CSF) samples were obtained at regular intervals and immunospecific IgM, immunoglobulin G (IgG) were quantified by ELISA while IL-6 was assayed using a cytometric bead array (CBA) kit. Results showed that following infection, CSF IgM, IgG, IL-6 and serum IL-6 were significantly (p0.05) between infection and 35 dpi but levels increased significantly (P<0.05) with the highest level of 55.53pg/ml recorded at112 dpi. IL-6 elevation from 35 dpi may be indicative of parasite neuroinvasion hence can be used as possible candidate marker for late stage disease in the monkey model. Further, the marker can also be used in conjunction with IgG and IgM as markers for development of test of cure for HAT. Copyright © 2017. Published by Elsevier B.V.

  10. Association between false-positive TORCH and antiphospholipid antibodies in healthy pregnant women.

    Science.gov (United States)

    De Carolis, S; Tabacco, S; Rizzo, F; Perrone, G; Garufi, C; Botta, A; Salvi, S; Benedetti Panici, P; Lanzone, A

    2018-04-01

    The present study investigated: (a) the presence of antiphospholipid antibodies and (b) the obstetric outcome in healthy pregnant women showing false-positive TORCH-Toxoplasmosis, Other: syphilis, varicella-zoster, Rubella, Cytomegalovirus (CMV), and Herpes infections-results. Data from 23 singleton healthy pregnancies with false-positive TORCH results were collected. Each woman was systematically screened for TORCH IgG and IgM during the pre-conception assessment and/or at the beginning of pregnancy. In the presence of IgM positivity, when indicated (CMV, toxoplasmosis, rubella, herpes simplex virus), IgG avidity was evaluated and, if possible, polymerase chain reaction was performed on an amniotic fluid sample in order to distinguish between primary infection or false positivity. The antiphospholipid antibodies tests were: lupus anticoagulant, anticardiolipin antibodies IgG, IgM, and anti-β2glicoprotein I IgG, IgM. The antiphospholipid antibodies tests, if positive, were repeated after 12 weeks to confirm the results. In pregnant women with false-positive TORCH, the overall prevalence of positive antiphospholipid antibodies for one or more tests was 52.2%. To clarify the correlation of false-positive TORCH results with clinical practice, obstetric outcome was analyzed in terms of live births, week of delivery, neonatal birth weight, and neonatal birth weight percentile. A statistically significant lower neonatal birth weight and neonatal birth weight percentile were observed in women with false-positive TORCH associated with antiphospholipid antibodies positivity (Group A) in comparison with those in women with false-positive TORCH without antiphospholipid antibodies positivity (Group B). No statistically significant difference was found for the week of delivery between the two groups. It is hoped that future studies will verify the life-long persistence of antiphospholipid antibodies positivity by follow-up of these women and identify who will develop a

  11. Indirect immunofluorescence(IgG and IgM tests for toxoplasmosis on 203 persons, with no symptomatology suggesting the disease, located in the city of Rio de Janeiro. Serological follow up one to two years later.

    Directory of Open Access Journals (Sweden)

    Maria Regina Reis Amendoeira

    1981-12-01

    Full Text Available Clinical and serological follow up examinations were performed on 203 persons, from three to twenty years of age, from the otolaryngology department of a hospital in the city of Rio de Janeiro, with no symptomatology suggesting toxoplasmosis, but suffering from chronic tonsillitis. According to results obtained during the first indirect immunofluorescence tests, the patients were divided into following groups: Group I (non-reactive IgG and IgM, 98 persons (48.3%; Group II (1:16 ≤ IgG ≤ 1:256 and non-reactive IgM, 74 persons (36.5%; Group III (IgM ≥ 1:1024 and non-reactive IgM, 18 persons (8.8%, and Group IV (IgG and IgM reactive, 13 persons (6.4%. One to two years later, 131 (64.5% of the 203 persons were reexamined by a second indirect immunofluorescence test. In the case of 66 persons (Group I whose serum was non-reactive in the IgG and IgM classes during the first indirect immunofluorescence test, serum conversion was observed in aproximately 21.2%. in 65 individuals (49.6%, (Groups II, III and IV,with reactive serum in the IgG classes during the first indirect immunofluorescence test, the second reaction showed an increase in titres in 20% of the cases, a decrease in 67.7% of the cases, or no alterations in 12.3 of the cases. In the IgM class, all 131 sera were non-reactive at 116 dilution the second immunofluorescence test, including the 13 cases that had previously been reactive in the immunoglobulin class, Symptomatology suggesting toxoplasmosis was only observed in one case during the second testing, this patient's principal physical sign being hypertrophied lymph nodes. during this period, the Toxoplasma antibodies showed titres of IgG 1:32000 and non-reactive IgM, whilst one year previously, during the first test, these titres were IgG 1:1024 and IgM 1:64. Differences in the age, sex and skin coloring of patients were not statistically significant as regards alterations in the indirect immunofluorescence test titres.Foi feito o

  12. Assessment of Anti-Tumor Cytotoxic Activity of Naturally Occurring Antibodies in Human Serum or Plasma.

    Science.gov (United States)

    Schwartz-Albiez, Reinhard; Dill, Othmar

    2017-01-01

    A small percentage of the Western population carries antibodies in the peripheral blood, which are able to kill human tumors such as neuroblastoma or melanoma. Several observations indicate that these antibodies, preferentially of IgM isotype, belong to the class of naturally occurring antibodies. Here, we describe two screening methods for the detection and quantification of such antibodies in human blood samples: a cellular ELISA technique and a flow cytometric assay, based on intercalation of fluorescent propidium iodide into the DNA of dying or dead cells.

  13. Titration of serum anti-ganglioside antibodies in patients with chronic medular injury previous to treatment with GM1 ganglioside

    Directory of Open Access Journals (Sweden)

    Barros Filho Tarcísio Eloy Pessoa

    2003-01-01

    Full Text Available Anti-ganglioside serum titers were evaluated by ELISA in 150 patients with complete spinal cord lesion for 6 to 12 months (IgG monosialo GM1, IgM monosialo GM1, IgG asialo GM1, IgM asialo GM1, IgG disialo GD1b e IgM disialo GD1b prior to treatment with GM1 100 mg/day i.m. Only 4 patients showed positive titers for anti-asialo-GM1 (IgM antibodies . All patients were clinically examined during and after treatment. No important side effects were observed with GM1 therapy. These results suggest that GM1-ganglioside administration in patients with chronic spinal cord injury is safe.

  14. The Molecular Determinants of Antibody Recognition and Antigenic Drift in the H3 Hemagglutinin of Swine Influenza A Virus

    Science.gov (United States)

    Abente, Eugenio J.; Santos, Jefferson; Lewis, Nicola S.; Gauger, Phillip C.; Stratton, Jered; Skepner, Eugene; Rajao, Daniela S.

    2016-01-01

    ABSTRACT Influenza A virus (IAV) of the H3 subtype is an important respiratory pathogen that affects both humans and swine. Vaccination to induce neutralizing antibodies against the surface glycoprotein hemagglutinin (HA) is the primary method used to control disease. However, due to antigenic drift, vaccine strains must be periodically updated. Six of the 7 positions previously identified in human seasonal H3 (positions 145, 155, 156, 158, 159, 189, and 193) were also indicated in swine H3 antigenic evolution. To experimentally test the effect on virus antigenicity of these 7 positions, substitutions were introduced into the HA of an isogenic swine lineage virus. We tested the antigenic effect of these introduced substitutions by using hemagglutination inhibition (HI) data with monovalent swine antisera and antigenic cartography to evaluate the antigenic phenotype of the mutant viruses. Combinations of substitutions within the antigenic motif caused significant changes in antigenicity. One virus mutant that varied at only two positions relative to the wild type had a >4-fold reduction in HI titers compared to homologous antisera. Potential changes in pathogenesis and transmission of the double mutant were evaluated in pigs. Although the double mutant had virus shedding titers and transmissibility comparable to those of the wild type, it caused a significantly lower percentage of lung lesions. Elucidating the antigenic effects of specific amino acid substitutions at these sites in swine H3 IAV has important implications for understanding IAV evolution within pigs as well as for improved vaccine development and control strategies in swine. IMPORTANCE A key component of influenza virus evolution is antigenic drift mediated by the accumulation of amino acid substitutions in the hemagglutinin (HA) protein, resulting in escape from prior immunity generated by natural infection or vaccination. Understanding which amino acid positions of the HA contribute to the ability

  15. Dengue antibodies in blood donors Pesquisa de anticorpos antidengue em doadores de sangue

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    Rejane Cristina Ribas-Silva

    2012-01-01

    Full Text Available BACKGROUND: Dengue is an urban arbovirus whose etiologic agent is a virus of the genus Flavorius with four distinct antigen serotypes (DENV-1, DENV-2, DENV-3 and DENV-4 that is transmitted to humans through the bite of the mosquito Aedes aegypti. The Campo Mourão region in Brazil is endemic for dengue fever. OBTECTIVE: The aim of this study was to evaluate the presence of IgG and IgM antibodies specific to the four serotypes of dengue in donors of the blood donor service in the city of Campo Mourão. METHODS: Epidemiological records were evaluated and 4 mL of peripheral blood from 213 blood donors were collected in tubes without anticoagulant. Serum was then obtained and immunochromatographic tests were undertaken (Imuno-Rápido Dengue IgM/IgG TM. Individuals involved in the study answered a social and epidemiological questionnaire on data which included age, gender and diagnosis of dengue. RESULTS: Only three (1.4% of the 213 blood tests were positive for IgG anti-dengue antibodies. No donors with IgM antibody, which identifies acute infection, were identified. CONCLUSIONS: The results of the current analysis show that the introduction of quantitative or molecular serological methods to determine the presence of anti-dengue antibodies or the detection of the dengue virus in blood donors in endemic regions should be established so that the quality of blood transfusions is guaranteed.OBJETIVO: O objetivo desta pesquisa foi realizar uma triagem sorológica para detectar a presença de anticorpos IgG e IgM específicos para os quatro sorotipos da dengue em doadores de sangue atendidos pelo Hemonúcleo de Campo Mourão-PR. MÉTODOS: foram aplicadas fichas epidemiológicas e coletados 4 mL de sangue periférico em tubos sem anticoagulante de 213 doadores de sangue. Posteriormente, foi obtido o soro e realizado o teste imunocromatrográfico (Imuno - Rápido Dengue IgM/IgG. Os indivíduos envolvidos neste estudo responderam um questionário s

  16. Measurement of Levels of Ebstein-Barr Virus Antibodies in Patients with Hodgkins Lymphoma and Comparison with Normal Population

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    M Mortazavi-zadeh

    2004-07-01

    Full Text Available Introduction: Hodgkins lymphoma is a unique malignancy with unknown etiology .Curability and prognosis of Hodgkin,s disease (HD depends on quickly early diagnosis .One of hypothesis proposed for the cause of this disease is Epstein- Barr virus infection and its activity in HD patients . Material and Methods:This case- control study was performed to determine the type and titers of antibodies against EBV capsid Antigens (Anti VCA IgM & IgG in HD patients as compared to the general population and its relation to age , sex , and subtype of Hodgkin. Thus, a fifty- person group of Hodgkin disease patients as the case group and a fifty – person group from the general population with the same age and sex characteristics as the control group were studied. Result: There was no significant difference for mean titer of IgM between two age ranges in each group of case and control. Also, there was statistically no significant difference between case and control groups ( P.Value=0.558 .Most of the patients as well as non affected persons had negative IgM titers. Regarding IgG, there was statistically no significant difference between case and control groups for being either negative or positive, and most persons (92% of each group and were positive for IgG, but mean titer of IgG was 2.87 mmol/lit in case group and 1.50 mmol/lit in control group , and this difference between two groups was statistically significant (Pvalue = 0.0001 . Conclusion: High titer of Anti-VCA IgG in Hodgkin disease patients compared to general population as seen in this study can explain over activity of EBV in Hodgkin's disease patients and the probable role of EBV in establishment and/or activity of the disease.

  17. Histone H1(0) mapping using monoclonal antibodies.

    Science.gov (United States)

    Dousson, S; Gorka, C; Gilly, C; Lawrence, J J

    1989-06-01

    Monoclonal antibodies (mAb) to ox liver histone H1 degree were produced and characterized. Two sets of mice were immunized either with pure H1(0) or with an H1(0)-yeast tRNA complex. Eleven hybridomas of various clonal origin were selected. Typing of the antibodies indicated that all but three IgM belonged to the IgG1 class and contained kappa light chains. Immunoblotting experiments using peptides derived from H1(0) or H5 treated by various proteolytic agents (trypsin, N-bromosuccinimide, cyanogen bromide, acetic acid), revealed that nine of the mAb reacted with the globular part of H1(0). More advanced characterization of the antigenic determinants allowed us to determine distinct regions within this globular part which are involved in the antigenic recognition. The peptopes could be subdivided into two groups. Three mAb bound to residues 24-27 and were specific for H1(0). Six mAb bound to residues 27-30 and were specific for H1(0) except one of them which strongly cross-reacted with H5 and GH5. Two mAb reacted with the entire histone H1(0) but failed to react with any of the peptides, suggesting that the corresponding epitope is a conformational antigenic determinant. In order to confirm the localization of the two distinct regions which are involved in the antigenic recognition, a synthetic decapeptide corresponding to the beginning of human H1(0) globular part (from residue 19 to residue 28) was synthesized. Inhibition experiments of the reaction between H1(0) and the various IgG1 mAb by increasing amounts of peptide-bovine serum albumin conjugates were then performed.

  18. Mechanisms underlying acquired von Willebrand syndrome associated with an IgM paraprotein.

    Science.gov (United States)

    Mayerhofer, M; Haushofer, A; Kyrle, P A; Chott, A; Müllner, C; Quehenberger, P; Worel, N; Traby, L; Eichinger, S

    2009-09-01

    Acquired von Willebrand (vW) syndrome is a rare bleeding disorder which is frequently associated with immunological, malignant or cardiovascular disorders. The underlying pathomechanisms, particularly in patients with IgM monoclonal gammopathies, often remain unknown. We report a patient with indolent small B-cell lymphoma (immunocytoma) and plasmacytic differentiation with an IgM kappa paraprotein who was admitted with retroperitoneal haematoma. Medical history and coagulation testing were consistent with acquired vW syndrome. vW immunohistochemistry showed normal cytoplasmic labelling of endothelial cells and megakaryocytes, whereas the lymphomatous infiltrate was negative. Acquired vW syndrome due to adsorption of vW factor on malignant cells was thus excluded. In the multimeric analysis, all multimers were present similar to that in type 1 vW syndrome, but the triplet structures were blurred. The bands on serum immunofixation electrophoresis were also atypically broadened, which suggested complex formation between the IgM and vW factor. Immunoprecipitation studies showed that the 176-kDa proteolytic fragment of vW factor co-precipitated with the IgM paraprotein in the patient but not in the controls, suggesting a specific interaction between vW factor and the paraprotein in the patient. The patient required surgery and was successfully managed by chemotherapy consisting of rituximab and fludarabin as well as plasma exchange.

  19. The sensitivity and specificity of Lassa virus IgM by ELISA as ...

    African Journals Online (AJOL)

    RT‑PCR) test, the gold standard for diagnosis, is unavailable in most centers. Serologic detection of LAV IgM is a more accessible tool and this work was to investigate its adequacy as an early marker for LF. Patients and Methods: A prospective ...

  20. IGMS: An Integrated ISO-to-Appliance Scale Grid Modeling System

    Energy Technology Data Exchange (ETDEWEB)

    Palmintier, Bryan; Hale, Elaine; Hansen, Timothy M.; Jones, Wesley; Biagioni, David; Sorensen, Harry; Wu, Hongyu; Hodge, Bri-Mathias

    2017-05-01

    This paper describes the Integrated Grid Modeling System (IGMS), a novel electric power system modeling platform for integrated transmission-distribution analysis that co-simulates off-the-shelf tools on high performance computing (HPC) platforms to offer unprecedented resolution from ISO markets down to appliances and other end uses. Specifically, the system simultaneously models hundreds or thousands of distribution systems in co-simulation with detailed Independent System Operator (ISO) markets and AGC-level reserve deployment. IGMS uses a new MPI-based hierarchical co-simulation framework to connect existing sub-domain models. Our initial efforts integrate opensource tools for wholesale markets (FESTIV), bulk AC power flow (MATPOWER), and full-featured distribution systems including physics-based end-use and distributed generation models (many instances of GridLAB-D[TM]). The modular IGMS framework enables tool substitution and additions for multi-domain analyses. This paper describes the IGMS tool, characterizes its performance, and demonstrates the impacts of the coupled simulations for analyzing high-penetration solar PV and price responsive load scenarios.

  1. Prevalence of Toxoplasma gondii IgG and IgM and associated risk ...

    African Journals Online (AJOL)

    Prevalence of Toxoplasma gondii IgG and IgM and associated risk factors among HIV-positive and HIV-negative patients in Vhembe district of South Africa. ... shown a high prevalence of T. gondii (IgG) among patients attending different HIV clinics in the Vhembe district with no current infections among pregnant women.

  2. Truly selective primary IgM deficiency is probably very rare

    NARCIS (Netherlands)

    Janssen, L.M.A.; Macken, T.; Creemers, M.C.W.; Pruijt, J.F.M.; Eijk, J.J.J.; de Vries, E.

    2017-01-01

    Isolated decreased serum-immunoglobulin (Ig)M has been associated with severe and/or recurrent infections, atopy and autoimmunity. However, the reported high prevalence of clinical problems in IgM-deficient patients may reflect the skewed tertiary centre population studied so far. Also, many papers

  3. Immunoglobulin classes (IgG, IgA and IgM) and acute phase ...

    African Journals Online (AJOL)

    The results indicate that USS or pregnancy changes different aspects of humoral immunity, thus the co-existence of pregnancy and S. haematobium infection may ... Résultats et conclusions: IgG, IgA et IgM étaient remarquablement élevés chez les femmes enceintes atteintes de la schistose urinaire par rapport aux femmes ...

  4. Serologic Survey of Specific Rubella Virus IgM in the Sera of ...

    African Journals Online (AJOL)

    Erah

    1.64%) were positive for Rubella IgM ... antenatal screening and vaccination of all females of child bearing age to eliminate this potentially devastating virus in the county (Afr J ... and July 2007 and transported on ice in cold boxes to the Virology ...

  5. Scalp Lesions in a Pediatric Patient with Hyper IgM Syndrome: Clinical and Histologic Mimicry of Cryptococcus neoformans Infection.

    Science.gov (United States)

    Acker, Karen P; Fetch, Audrey; Schnell, Stephanie A; Hammond, Jennifer; Herrera, Christina; Niedt, George; Ratner, Adam J; Lauren, Christine T

    2018-01-01

    We report a case of cutaneous cryptococcosis due to Cryptococcus neoformans in a pediatric patient with hyper IgM syndrome with scalp lesions that resembled tinea capitis on gross examination and mimicked juvenile xanthogranuloma on histologic examination. This case highlights the importance of considering cutaneous cryptococcosis in patients with hyper IgM syndrome. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Comparison of in-house IgM and IgG ELISAs for the serodiagnosis of melioidosis in Malaysia.

    Science.gov (United States)

    Hii, Shirley Yi Fen; Ali, Noor Azila; Ahmad, Norazah; Amran, Fairuz

    2017-11-01

    Melioidosis is an endemic infectious disease in Southeast Asia and northern Australia, caused by Burkholderia pseudomallei. However, the incidence rate in Malaysia is not well documented. The high mortality rate and broad range of clinical presentations require rapid and accurate diagnosis for appropriate treatment. This study compared the efficacy of in-house IgM and IgG ELISA methods using a local B. pseudomallei strain. The diagnostic accuracy of the in-house IgG ELISA was better than that of the IgM ELISA: sensitivity (IgG: 84.71 %, IgM: 76.14 %) and specificity (IgG: 93.64 %, IgM: 90.17 %); positive predictive value (IgG: 86.75 %, IgM: 79.76 %) and negative predictive value (IgG: 92.57 %, IgM: 89.66 %); likelihood ratio (LR) [IgG: 13.32, IgM: 7.75 (LR+); IgG: 0.16, IgM: 0.26 (LR-)], and was supported by the observation of the absorbance value in comparisons between culture and serology sampling. In-house IgG ELISA was shown to be useful as an early diagnostic tool for melioidosis.

  7. Rapid diagnosis of occult abscesses using sup 99m Tc-labeled monoclonal antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Coons, T.A.; Rhodes, B.A. (RhoMed, Inc., Albuquerque, NM (USA)); Thakur, M.L. (Thomas Jefferson Univ., Philadelphia, PA (USA)); Marcus, C.S. (Harbor-UCLA Medical Center, Torrance, CA (USA)); Ballou, B. (Pittsburgh Univ., PA (USA))

    1991-01-01

    Acute infections, such as appendicitis and occult infections in AIDS patients, can be diagnosed within two hours by gamma scintigraphy after i.v. administration of {sup 99m}Tc labeled antibodies reactive with human granulocytes. The antibody, murine IgM anti-SSEA-1, is partially reduced using Sn(II) to expose and protect reactive sulfide groups. The antibody is then purified, stannous tartrate and stabilizers are added, and the mixture is lyophilized. To label, sodium pertechnetate is added. After a 15 minute incubation the tracer drug is injected. The rate of accumulation and degree of concentration at the site of infection is presumptively determinative of the severity of the infection. Acceptance criteria and tests for the {sup 99m}Tc labeled antibody product have been established and validated. Greater than 93% of the {sup 99m}Tc is firmly bound to the protein as determined by quantitative HPLC. Radiochemical impurities, colloidal {sup 99m}Tc and free pertechnetate are together less than 4% as determined by thin layer chromatography. The immunoreactive fraction, measured by binding to solid phase antigen, and affinity measured be ELISA, are unchanged by the {sup 99m}Tc-direct labeling process. Two hour blood clearance in rats is within 90% of the value of the {sup 125}I labeled analog. The immunoreactive fraction decreases less than 10% when incubated in human plasma for 24 hours. This method has been compared to other direct labeling methods, and found to give higher radiochemical yields. 5 figs.

  8. Molecular determinants of dengue virus 2 envelope protein important for virus entry in FcγRIIA-mediated antibody-dependent enhancement of infection

    Energy Technology Data Exchange (ETDEWEB)

    Chotiwan, Nunya; Roehrig, John T. [Arboviral Diseases Branch, Division of Vector-Borne Disease, Centers for Disease Control and Prevention, Fort Collins, CO 80521 (United States); Schlesinger, Jacob J. [Department of Medicine, University of Rochester, Rochester, NY 14642 (United States); Blair, Carol D. [Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523 (United States); Huang, Claire Y.-H., E-mail: yxh0@cdc.gov [Arboviral Diseases Branch, Division of Vector-Borne Disease, Centers for Disease Control and Prevention, Fort Collins, CO 80521 (United States)

    2014-05-15

    Antibody-dependent enhancement (ADE) of infection may cause severe illness in patients suffering a secondary infection by a heterologous dengue virus (DENV) serotype. During ADE of infection, cross-reactive non- or poorly-neutralizing antibodies form infectious virus-Ab complexes with the newly infecting serotype and enhance virus infection by binding to the Fcγ receptors (FcγR) on FcγR-bearing cells. In this study, we determined that molecular determinants of DENV2 envelope protein critical for virus entry during non-ADE infection are also required for ADE infection mediated by FcγRIIA, and binding of virus-Ab complexes with FcγRIIA alone is not sufficient for ADE of infection. The FcγRIIA mainly plays an auxiliary role in concentrating the virus–Ab complex to the cell surface, and other primary cellular receptors are required for virus entry. Understanding the viral entry pathway in ADE of DENV infection will greatly facilitate rational designs of anti-viral therapeutics against severe dengue disease associated with ADE. - Highlights: • KKK305/307/310 in DENV2 E-DIII is critical for virus attachment in ADE and non-ADE infection. • Binding of DENV2–Ab complex with FcγRII alone is not sufficient for virus entry in ADE infection. • Other primary receptors were required for DENV2 internalization during FcγRII–mediated ADE. • G104 and L135 of DENV2 E are critical for virus-mediated membrane fusion. • DENV2 virus-mediated membrane fusion is required for both ADE and non-ADE infection.

  9. Molecular determinants of dengue virus 2 envelope protein important for virus entry in FcγRIIA-mediated antibody-dependent enhancement of infection

    International Nuclear Information System (INIS)

    Chotiwan, Nunya; Roehrig, John T.; Schlesinger, Jacob J.; Blair, Carol D.; Huang, Claire Y.-H.

    2014-01-01

    Antibody-dependent enhancement (ADE) of infection may cause severe illness in patients suffering a secondary infection by a heterologous dengue virus (DENV) serotype. During ADE of infection, cross-reactive non- or poorly-neutralizing antibodies form infectious virus-Ab complexes with the newly infecting serotype and enhance virus infection by binding to the Fcγ receptors (FcγR) on FcγR-bearing cells. In this study, we determined that molecular determinants of DENV2 envelope protein critical for virus entry during non-ADE infection are also required for ADE infection mediated by FcγRIIA, and binding of virus-Ab complexes with FcγRIIA alone is not sufficient for ADE of infection. The FcγRIIA mainly plays an auxiliary role in concentrating the virus–Ab complex to the cell surface, and other primary cellular receptors are required for virus entry. Understanding the viral entry pathway in ADE of DENV infection will greatly facilitate rational designs of anti-viral therapeutics against severe dengue disease associated with ADE. - Highlights: • KKK305/307/310 in DENV2 E-DIII is critical for virus attachment in ADE and non-ADE infection. • Binding of DENV2–Ab complex with FcγRII alone is not sufficient for virus entry in ADE infection. • Other primary receptors were required for DENV2 internalization during FcγRII–mediated ADE. • G104 and L135 of DENV2 E are critical for virus-mediated membrane fusion. • DENV2 virus-mediated membrane fusion is required for both ADE and non-ADE infection

  10. Multi-scale calculation and global-fit analysis of hydrodynamic properties of biological macromolecules: determination of the overall conformation of antibody IgG molecules.

    Science.gov (United States)

    Amorós, D; Ortega, A; Harding, S E; García de la Torre, J

    2010-02-01

    We present a scheme, based on existing and newly developed computational tools, for the determination of the overall conformation of biological macromolecules composed by domains or subunits, using from such structural determination easily available solution properties. In a multi-scale approach, atomic-level structures are used to provide simple shapes for the subunits, which are put together in a coarse grained model, with a few parameters that determine the overall shape of the macromolecule. Computer programs, like those in the HYDRO suite that evaluate the properties of either atomic or coarse-grained models. In this paper we present a new scheme for a global fit of multiple properties, implemented in a new computer program, HYDROFIT, which interfaces with the programs of the HYDRO suite to find an optimum, best-fitting structure in a robust but simple way. The determination of the overall structure of the native antibody IgG3, bearing a long hinge, and that of the hingeless mutant m15 is presented to test and confirm the validity of this simple, systematic and efficient scheme.

  11. Human antibody recognition of xenogeneic antigens (NeuGc and Gal) on porcine heart valves: could genetically modified pig heart valves reduce structural valve deterioration?

    Science.gov (United States)

    Lee, Whayoung; Long, Cassandra; Ramsoondar, Jagdeece; Ayares, David; Cooper, David K C; Manji, Rizwan A; Hara, Hidetaka

    2016-09-01

    Glutaraldehyde-fixed bioprosthetic heart valves (GBHVs) derived from wild-type (WT, genetically unmodified) pigs are widely used clinically for heart valve replacement. There is evidence that their failure is related to an immune response. The use of valves from genetically engineered pigs that do not express specific pig antigens may prolong GBHV survival. Our aims were to determine (i) expression of Gal and NeuGc on heart (aortic and pulmonary) valves and pericardium of WT, α1,3-galactosyltransferase gene knockout (GTKO) and GTKO/N-glycolylneuraminic acid gene-knockout (GTKO/NeuGcKO) pigs in comparison with three different commercially available GBHVs and (ii) to determine human antibody binding to these tissues. Wild-type, GTKO/CD46, and GTKO/CD46/NeuGcKO pig valves and pericardium were tested (i) fresh and (ii) after fixation with glutaraldehyde (0.02%, 0.2%, 2%). Sections of GBHVs, fresh and fixed valves, and pericardium were stained for Gal and NeuGc expression, and for human IgM and IgG antibody binding. Gal and NeuGc expression was high on all GBHVs and WT pig valves/pericardium, but was absent after antigen-specific-knockout. There was no difference in antigen expression or antibody binding among WT aortic, pulmonary valves, and pericardium as well as GBHVs. Glutaraldehyde fixation did not alter expression of Gal or NeuGc. After incubation with human serum, human IgM and IgG bound to all GBHVs and WT pig valves/pericardium. Valves from GTKO/CD46 pigs and, particularly, GTKO/CD46/NeuGcKO pigs (with/without glutaraldehyde fixation) showed less IgM and IgG binding. Compared to WT pigs, GTKO/CD46/NeuGcKO pigs would be preferable sources of GBHVs, because the absence of Gal/NeuGc expression reduces human antibody binding. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  12. IgG/IgM antiphospholipid antibodies present in the classification criteria for the antiphospholipid syndrome: a critical review of their association with thrombosis.

    Science.gov (United States)

    Kelchtermans, H; Pelkmans, L; de Laat, B; Devreese, K M

    2016-08-01

    Essentials The clinical value of IgM antibodies in thrombotic antiphospholipid syndrome (APS) is debated. By review of literature, we reconsidered the clinical value of IgM antibodies in thrombotic APS. More significant correlations with thrombosis were found for the IgG compared to IgM isotype. Unavailability of paired IgG/IgM results hampers evaluating the added value of IgM positivity. Click to hear Dr de Groot's perspective on antiphospholipid syndrome Background Despite the update of the classification criteria for the antiphospholipid syndrome (APS), difficulties persist in the identification of patients at risk for thrombosis. Current guidelines include assays detecting IgG/IgM anti-β2 -glycoprotein I and anti-cardiolipin antibodies, although the relevance of IgM antibodies has been debated. Objectives Through a review of the literature from 2001 to 2014, we aimed to formally establish the thrombotic risk stratification potential of IgM as compared with IgG anti-phospholipid antibodies (aPLs). Patients/methods One thousand two hundred and twenty-eight articles were selected by a computer-assisted search of the literature. Of the 177 studies that met our inclusion criteria, the clinical value of IgG/IgM aPLs was established through analysis of odds ratios for thrombosis or percentage of positives in the thrombotic population. Results/conclusions We clearly found more significant correlations with thrombosis for the IgG than for the IgM isotype. Nonetheless, in a minority of studies, significant associations with thrombosis were found for IgM but not IgG antibodies. The unavailability of paired results of IgG and IgM for each separate patient hampers evaluation of the added value of isolated IgM positivity. To fully take advantage of results obtained by future studies, we strongly encourage scientists to provide all studied information per patient. We planned a large multicenter study to investigate clinical associations of isolated/combined positivity for

  13. Anticorpos antifosfolípides em mulheres com antecedentes de perdas gestacionais: estudo caso-controle Antiphospholipid antibodies in women with recurrent pregnancy loss: a case-control study

    Directory of Open Access Journals (Sweden)

    Olívia Lúcia Nunes Costa

    2005-06-01

    Full Text Available OBJETIVO: determinar a prevalência de anticorpos antifosfolípides em mulheres com antecedentes de perdas gestacionais na população obstétrica em geral e verificar se os anticorpos antifosfolípides representam fator de risco para perdas gestacionais na população estudada. MÉTODOS: foi realizado um estudo caso-controle prospectivo com mulheres grávidas e não grávidas, atendidas numa maternidade pública entre março de 2003 e junho de 2004. As mulheres foram divididas em dois grupos de acordo com o passado obstétrico; 100 mulheres com antecedentes de perdas gestacionais de acordo com a definição estabelecida para o diagnóstico da síndrome antifosfolipídica e que não apresentassem outros fatores relacionados ao insucesso gestacional; 150 mulheres saudáveis com antecedentes de duas ou mais gestações bem sucedidas. A determinação do anticoagulante lúpico (AL foi feita mediante os testes de TTPA, dRVVT de triagem e dRVVT confirmatório. Para a pesquisa dos anticorpos anticardiolipina (aCL classes IgG e IgM foi utilizado o teste de ELISA. e os resultados semiquantitativos expressos em unidades GPL e MPL. RESULTADOS: o anticoagulante lúpico estava presente em 5% das pacientes-caso e 2% dos controles (p=0,27. Os anticorpos aCL IgG estavam presentes em 18% das pacientes-caso e 8,7% das pacientes-controle (p=0,028; OR=2,3; IC 95%=1-53. Na classe IgM, 5% de positividade para os casos e 1% para os controles (p=0,21. CONCLUSÕES: os anticorpos antifosfolípides (AL e/ou aCL IgG e/ou IgM foram mais prevalentes nas mulheres com perdas gestacionais (28% que na população obstétrica em geral (17%. As mulheres com aCL IgG têm duas vezes mais chance de ter perdas gestacionais que a população obstétrica em geral.OBJECTIVE: To determine the prevalence of antiphospholipid antibodies in women with pregnancy loss and verify if such antibodies represent a risk factor for pregnancy failure. METHODS: We performed a case-control study with

  14. Avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii: immunological marker for acute recent toxoplasmosis Avidez de anticorpos IgG anti-antígeno de secreção e excreção de Toxoplasma gondii: marcador imunológico de toxoplasmose aguda recente

    Directory of Open Access Journals (Sweden)

    Patrícia Regina Barboza Araújo

    2008-04-01

    Full Text Available Detection of anti-toxoplasma IgM antibodies has frequently been used as a serological marker for diagnosing recently acquired toxoplasmosis. However, the persistence of these antibodies in some patients has complicated the interpretation of serological results when toxoplasmosis is suspected. The purpose of the present study was to evaluate the avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii by means of immunoblot, to establish a profile for acute recent infection in a single serum sample and confirm the presence of residual IgM antibodies obtained in automated assays. When we evaluated the avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii by means of immunoblot, we observed phase-specific reactivity, i.e. cases of acute recent toxoplasmosis presented low avidity and cases of non-acute recent toxoplasmosis presented high avidity towards the 30kDa protein fraction, which probably corresponds to the SAG-1 surface antigen. Our results suggest that the avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii is an important immunological marker for distinguishing between recent infections and for determining the presence of residual IgM antibodies obtained from automated assays.A detecção de anticorpos IgM antitoxoplasma tem sido freqüentemente utilizada como marcador sorológico para o diagnóstico de toxoplasmose de aquisição recente. Entretanto, a persistência destes anticorpos em alguns pacientes tem complicado a interpretação dos resultados sorológicos quando a toxoplasmose é suspeitada. A proposta deste trabalho foi avaliar a avidez de anticorpos IgG contra antígenos de secreção e excreção de Toxoplasma gondii por immunoblot, para estabelecer um perfil de infecção recente aguda em uma única amostra de soro e confirmar a presença de anticorpos IgM residuais obtidos nos testes automatizados. Quando a avidez de anticorpos IgG contra ant

  15. Heterogeneity of Polyneuropathy Associated with Anti-MAG Antibodies

    Directory of Open Access Journals (Sweden)

    Laurent Magy

    2015-01-01

    Full Text Available Polyneuropathy associated with IgM monoclonal gammopathy and anti-myelin associated glycoprotein (MAG antibodies is an immune-mediated demyelinating neuropathy. The pathophysiology of this condition is likely to involve anti-MAG antibody deposition on myelin sheaths of the peripheral nerves and it is supposed to be distinct from chronic inflammatory demyelinating neuropathy (CIDP, another immune-mediated demyelinating peripheral neuropathy. In this series, we have retrospectively reviewed clinical and laboratory findings from 60 patients with polyneuropathy, IgM gammopathy, and anti-MAG antibodies. We found that the clinical picture in these patients is highly variable suggesting a direct link between the monoclonal gammopathy and the neuropathy. Conversely, one-third of patients had a CIDP-like phenotype on electrodiagnostic testing and this was correlated with a low titer of anti-MAG antibodies and the absence of widening of myelin lamellae. Our data suggest that polyneuropathy associated with anti-MAG antibodies is less homogeneous than previously said and that the pathophysiology of the condition is likely to be heterogeneous as well with the self-antigen being MAG in most of the patients but possibly being another component of myelin in the others.

  16. Combination of SDS-PAGE and intact mass analysis for rapid determination of heterogeneities in monoclonal antibody therapeutics.

    Science.gov (United States)

    Yamada, Hideaki; Matsumura, Chiemi; Yamada, Keita; Teshima, Koichiro; Hiroshima, Takashi; Kinoshita, Mitsuhiro; Suzuki, Shigeo; Kakehi, Kazuaki

    2017-05-01

    mAbs are currently mainstream in biopharmaceuticals, and their market has been growing due to their high target specificity. Characterization of heterogeneities in mAbs is performed to secure their quality and safety by physicochemical analyses. However, they require time-consuming task, which often strain the resources of drug development in pharmaceuticals. Rapid and direct method to determine the heterogeneities should be a powerful tool for pharmaceutical analysis. Considering the advantages of electrophoresis and MS, this study addresses the combination of SDS-PAGE and intact mass analysis, which provides direct, rapid, and orthogonal determination of heterogeneities in mAb therapeutics. mAb therapeutics that migrated in SDS-PAGE were recovered from gel by treatment with SDC-containing buffer. Usage of SDC-containing buffer as extraction solvent and ethanol-based staining solution enhanced the recovery of intact IgG from SDS-PAGE gels. Recovery of mAbs reached more than 86% with 0.2% SD. The heterogeneities, especially N-glycan variants in the recovered mAb therapeutics, were clearly determined by intact mass analysis. We believe that the study is important in pharmaceuticals‧ perspective since orthogonal combination of gel electrophoresis and intact mass analysis should be pivotal role for rapid and precise characterization of mAbs. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Progression from IgD+ IgM+ to isotype-switched B cells is site specific during coronavirus-induced encephalomyelitis.

    Science.gov (United States)

    Phares, Timothy W; DiSano, Krista D; Stohlman, Stephen A; Bergmann, Cornelia C

    2014-08-01

    Various infections in the central nervous system (CNS) trigger B cell accumulation; however, the relative dynamics between viral replication and alterations in distinct B cell subsets are largely unknown. Using a glia-tropic coronavirus infection, which is initiated in the brain but rapidly spreads to and predominantly persists in the spinal cord, this study characterizes longitudinal changes in B cell subsets at both infected anatomical sites. The phase of T cell-dependent, antibody-independent control of infectious virus was associated with a similar recruitment of naive/early-activated IgD(+) IgM(+) B cells into both the brain and spinal cord. This population was progressively replaced by CD138(-) IgD(-) IgM(+) B cells, isotype-switched CD138(-) IgD(-) IgM(-) memory B cells (B(mem)), and CD138(+) antibody-secreting cells (ASC). A more rapid transition to B(mem) and ASC in spinal cord than in brain was associated with higher levels of persisting viral RNA and transcripts encoding factors promoting B cell migration, differentiation, and survival. The results demonstrate that naive/early-activated B cells are recruited early during coronavirus CNS infection but are subsequently replaced by more differentiated B cells. Furthermore, viral persistence, even at low levels, is a driving force for accumulation of isotype-switched B(mem) and ASC. Acute and chronic human CNS infections are associated with an accumulation of heterogeneous B cell subsets; however, their influence on viral load and disease is unclear. Using a glia-tropic coronavirus model, we demonstrate that the accumulation of B cells ranging from early-activated to isotype-switched differentiation stages is both temporally and spatially orchestrated. Acutely infected brains and spinal cords indiscriminately recruit a homogeneous population of early-activated B cells, which is progressively replaced by diverse, more differentiated subsets. The latter process is accelerated by elevated proinflammatory

  18. TOXOPLASMA AND VIRAL ANTIBODIES AMONG HIV PATIENTS AND INMATES IN CENTRAL JAVA, INDONESIA.

    Science.gov (United States)

    Sari, Yulia; Haryati, Sri; Raharjo, Irvan; Prasetyo, Afiono Agung

    2015-11-01

    In Indonesia, Toxoplasma and its associations with blood-borne viruses have been poorly studied. In order to study the association between anti-Toxoplasma antibodies and blood-borne viral antibodies, blood samples from 497 participants (375 inmates from four prisons in Central Java, Indonesia and 122 HIV patients at a Voluntary Counseling and Testing Clinic in Surakarta, Indonesia) were tested for serological markers of Toxoplasma, human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV) and human T-lymphotropic virus types I and II (HTLV-1/2). Anti-Toxoplasma IgG and IgM positivity rates were 41.6% and 3.6%, respectively. One point two percent of participants was positive for both anti-Toxoplasma IgG and IgM antibodies. Sixteen point five percent, 11.3%, 2.6% and 2.8% of participants were positive for anti- Toxoplasma IgG combined with anti-HCV antibodies, anti-Toxoplasma IgG combined with anti-HIV antibodies, anti-Toxoplasma IgM combined with anti-HIV antibodes and anti-Toxoplasma IgG combined with both anti-HIV and anti-HCV antibodies, respectively. Anti-Toxoplasma IgM seropositivity was associated with anti-HIV (aOR = 4.3; 95% CI: 1.112-16.204, p = 0.034). Anti-Toxoplasma IgG antibodies were associated with anti-HCV (aOR = 2.8; 95% CI: 1.749-4.538, p < 0.001) and history of injection drug use (aOR = 3.1; 95% CI: 1.905-5.093, p < 0.001). In conclusion, we recommend patients with HIV, HCV infection and injection drug users should be screened for Toxoplasma infection in Indonesia.

  19. Anti-beta2-glycoprotein I antibodies are associated with pregnancy loss in women with the lupus anticoagulant.

    Science.gov (United States)

    Sailer, Thomas; Zoghlami, Claudia; Kurz, Christine; Rumpold, Helmut; Quehenberger, Peter; Panzer, Simon; Pabinger, Ingrid

    2006-05-01

    The presence of lupus anticoagulant (LA) predisposes to fetal loss and to venous and arterial thrombosis; however, a subgroup of women is unaffected by pregnancy loss. Currently, no predictive markers are available for the identification of women positive for LA at increased risk for pregnancy loss. It was the aim of our study to investigate whether increased anti-beta2-GPI-antibodies predict pregnancy loss in women positive for LA. We performed a cross-sectional study in a cohort of 39 women with persistent LA, who had in total 111 pregnancies. Fifteen women had exclusively normal pregnancies (30 pregnancies) and 24 women had pregnancy losses (81 pregnancies). Anti-beta2-GPI-antibodies were determined using a semiquantitative enzyme linked immunoassay (QUANTA Lite beta2 GPI IgG and IgM; Inova Diagnostics). Increased levels of anti-beta2-GPI antibodies were significantly associated with pregnancy loss [odds ratio (OR) 9.6, 95% confidence interval (CI) 1.6-56.4]. This risk was even higher in the subgroup of women (n = 16) with more than two miscarriages or fetal loss after the first trimester [OR 13.1, 95% CI 1.4-126.3]. There was no significant association between anticardiolipin antibodies and pregnancy loss [OR 3.5, 95% CI 0.7-17.6]. The co-existence of anti-beta2-GPI and anticardiolipin antibodies was also predictive for pregnancy loss [OR 6.1, 95% CI 1.3-29.7]. Interestingly, the prevalence of thrombosis was similar between women with normal pregnancy (87%) and those with pregnancy loss (75%). We conclude that increased levels of anti-beta2-GPI antibodies are predictive for pregnancy loss among women positive for LA, and that prophylactic treatment should be considered in these women even without a history of previous pregnancy loss.

  20. Increased prevalence of diverse N-methyl-D-aspartate glutamate receptor antibodies in patients with an initial diagnosis of schizophrenia: specific relevance of IgG NR1a antibodies for distinction from N-methyl-D-aspartate glutamate receptor encephalitis.

    Science.gov (United States)

    Steiner, Johann; Walter, Martin; Glanz, Wenzel; Sarnyai, Zoltán; Bernstein, Hans-Gert; Vielhaber, Stefan; Kästner, Andrea; Skalej, Martin; Jordan, Wolfgang; Schiltz, Kolja; Klingbeil, Christine; Wandinger, Klaus-Peter; Bogerts, Bernhard; Stoecker, Winfried

    2013-03-01

    Evidence for symptomatic convergence of schizophrenia and N-methyl-D-aspartate glutamate receptor (NMDA-R) encephalitis highlights the need for an assessment of antibody prevalence and specificity for distinct disease mechanisms in patients with a diagnosis of schizophrenia among glutamatergic pathophysiologic abnormalities in psychiatric disorders. To compare the specificity and prevalence of NMDA-R antibodies in schizophrenia (DSM-IV criteria) with those of other psychiatric diagnoses and to determine whether antibody subtypes characterize overlap with and distinction from those in NMDA-R encephalitis. Serum from 459 patients admitted with acute schizophrenia, major depression (MD), and borderline personality disorder (BLPD) or individuals serving as matched controls was obtained from our scientific blood bank. To explore epitope specificity and antibody subtype, IgA/IgG/IgM NMDA-R (NR1a or NR1a/NR2b) and α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptors (AMPA-R) (GluR1/GluR2) serum antibodies were determined. Two hundred thirty matched healthy controls were compared with patients (unmedicated for at least 6 weeks) with schizophrenia (n = 121), MD (n = 70), or BLPD (n = 38). The primary outcome was the overall number of seropositive cases for NMDA-R and AMPA-R antibodies; the secondary outcome was disease specificity of IgA/IgG/IgM antibodies and epitope specificity for clinical subgroups. Diverse NMDA-R antibodies were identified in 15 subjects, primarily those with an initial schizophrenia diagnosis (9.9%), opposed to MD (2.8%), BLPD (0), and controls (0.4%). Retrospectively, 2 patients initially classified as having catatonic or disorganized schizophrenia were reclassified as having misdiagnosed NMDA-R encephalitis (presence of specific serum and cerebrospinal fluid IgG NR1a antibodies). In all other seropositive cases, the antibodies consisted of classes IgA and/or IgM or were directed against NR1a/NR2b (not against NR1a alone). None of the

  1. Determination of human IgG by solid substrate room temperature phosphorescence immunoassay based on an antibody labeled with nanoparticles containing dibromofluorescein luminescent molecules

    International Nuclear Information System (INIS)

    Liu Jiaming; Zhu Guohui; Rao Zhiming; Wei Changjin; Li Longdi; Chen Cuilian; Li Zhiming

    2005-01-01

    Luminescent silicon dioxide nano-particles with size of 20 nm, which containing dibromofluorescein (D) were synthesized by sol-gel method (symbolized by D-SiO 2 ).The particles can emit intense and stable room temperature phosphorescence signal on polyamide membrane when Pb(Ac) 2 was used as a heavy atom perturber. The λ exmax /λ emmax was 457/622 nm. Our research indicated that the specific immune reaction between goat-anti-human IgG antibody labeled with D-SiO 2 and human IgG could be carried out on polyamide membrane quantitatively, and the phosphorescence intensity of the particle was enhanced after the immunoreactions. Thus a new method of solid substrate room temperature phosphorescence immunoassay (SS-RTP-IA) for the determination of human IgG was established basing on antibody labeled with the D-SiO 2 nanoparticles. The linear range of this method was 0.0624-20.0 pg human IgG spot -1 (corresponding concentration: 0.156-50.0 ng ml -1 , the sample volume: 0.40 μl spot -1 ) with a limit of detection (LD) as 0.018 pg spot -1 , and the regression equation of working curve was ΔI p = 7.201 m IgG (pg spot -1 ) + 82.57. Samples containing 0.156 and 50.0 ng ml -1 of IgG were measured repeatedly for 11 times and R.S.D.s were 4.1 and 3.4%, respectively. Results showed that this method had the merits as sensitive, accurate and precise

  2. The value of antibody-coated bacteria in tracheal aspirates for the diagnosis of ventilator-associated pneumonia: a case-control study.

    Science.gov (United States)

    Ranzani, Otavio Tavares; Forte, Daniel Neves; Forte, Antonio Carlos; Mimica, Igor; Forte, Wilma Carvalho Neves

    2016-01-01

    Ventilator-associated pneumonia (VAP) is the leading type of hospital-acquired infection in ICU patients. The diagnosis of VAP is challenging, mostly due to limitations of the diagnostic methods available. The aim of this study was to determine whether antibody-coated bacteria (ACB) evaluation can improve the specificity of endotracheal aspirate (EA) culture in VAP diagnosis. We conducted a diagnostic case-control study, enrolling 45 patients undergoing mechanical ventilation. Samples of EA were obtained from patients with and without VAP (cases and controls, respectively), and we assessed the number of bacteria coated with FITC-conjugated monoclonal antibodies (IgA, IgM, or IgG) or an FITC-conjugated polyvalent antibody. Using immunofluorescence microscopy, we determined the proportion of ACB among a fixed number of 80 bacteria. The median proportions of ACB were significantly higher among the cases (n = 22) than among the controls (n = 23)-IgA (60.6% vs. 22.5%), IgM (42.5% vs. 12.5%), IgG (50.6% vs. 17.5%), and polyvalent (75.6% vs. 33.8%)-p 95,0% e > 93,3%, respectivamente. O número de BRA em amostras de AT foi maior nos casos que nos controles. Nossos achados indicam que a avaliação de BRA no AT é uma ferramenta promissora para aumentar a especificidade do diagnóstico de PAVM. A técnica pode ser custo-efetiva e, portanto, útil em locais com poucos recursos, com as vantagens de minimizar resultados falso-positivos e evitar o tratamento excessivo.

  3. Monoclonal antibodies for the measurement of class-specific antibody responses in the green turtle, Chelonia mydas.

    Science.gov (United States)

    Herbst, L H; Klein, P A

    1995-06-01

    Monoclonal antibodies (Mabs) were developed against the known immunoglobulin classes of the green turtle, Chelonia mydas. Plasma protein fractions enriched for 5.7S IgY, 7S IgY, and IgM turtle immunoglobulins were used to immunize Balb/c mice for hybridoma production and for hybridoma screening. Fifteen hybridomas produced Mabs with specificity for turtle immunoglobulins and for affinity purified dinitrophenol (DNP) specific turtle antibodies. Three Mabs specific for either turtle 5.7S IgY heavy chain (HL814), 7S IgY heavy chain (HL857), or IgM heavy chain (HL846) were purified and used in an enzyme-linked immunosorbent assay (ELISA) to measure antibody responses in two turtles immunized with 2,4-dinitrophenylated bovine serum albumin (DNP-BSA) over a 10 month period. In both turtles the 7S IgY antibody response developed within 5 weeks of the first inoculation and remained high over the following 9 months. The 5.7S IgY antibody response was detected in one turtle at 3-4 months and in the other at 8 months, and reached high levels in both individuals by 10 months. The IgM responses were difficult to interpret. One turtle had pre-inoculation anti-DNP IgM antibody in its plasma and the other developed only a weak, transient response at about 4 months. The class-specific antibody activity in immune turtle plasma could be strongly inhibited by soluble DNP or by rabbit anti-DNP specific antiserum, showing that these antibody responses were directed predominantly to the DNP hapten on the DNP-BSA antigen. Antibody responses to the BSA carrier could not be detected in either turtle over the course of the immunization. Mab HL814, specific for an epitope on the 5.7S green turtle immunoglobulin heavy chain, will be useful for characterizing the molecular relationships of 5.7S, 7S and IgM heavy chains and the role of 5.7S antibody in humoral immunity in this species. All anti-turtle Ig Mabs were screened against the plasma globulins of Loggerhead (Caretta caretta), Olive

  4. A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM

    International Nuclear Information System (INIS)

    Jung, Kyung oh; Youn, Hyewon; Kim, Seung Hoo; Kim, Young-Hwa; Kang, Keon Wook; Chung, June-Key

    2016-01-01

    Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and 64 Cu. HT29 (hTERT+) and U2OS (hTERT−) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus. In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo. - Highlights: • We developed new probes for imaging hTERT using Tat-conjugated IgM antibodies labeled with a fluorescent dye and radioisotope. • This probes could be used to overcome limitation of conventional antibody imaging system in live cell imaging. • This system could be applicable to monitor intracellular and intranuclear proteins in vitro and in vivo.

  5. A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Kyung oh [Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of); Biomedical Sciences, Seoul National University College of Medicine (Korea, Republic of); Cancer Research Institute, Seoul National University College of Medicine (Korea, Republic of); Tumor Microenvironment Global Core Research Center, Seoul National University (Korea, Republic of); Youn, Hyewon, E-mail: hwyoun@snu.ac.kr [Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of); Cancer Research Institute, Seoul National University College of Medicine (Korea, Republic of); Tumor Microenvironment Global Core Research Center, Seoul National University (Korea, Republic of); Cancer Imaging Center, Seoul National University Hospital, Seoul (Korea, Republic of); Kim, Seung Hoo [Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of); Cancer Research Institute, Seoul National University College of Medicine (Korea, Republic of); Kim, Young-Hwa [Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of); Biomedical Sciences, Seoul National University College of Medicine (Korea, Republic of); Cancer Research Institute, Seoul National University College of Medicine (Korea, Republic of); Kang, Keon Wook [Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of); Cancer Research Institute, Seoul National University College of Medicine (Korea, Republic of); Chung, June-Key, E-mail: jkchung@snu.ac.kr [Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of); Biomedical Sciences, Seoul National University College of Medicine (Korea, Republic of); Cancer Research Institute, Seoul National University College of Medicine (Korea, Republic of); Tumor Microenvironment Global Core Research Center, Seoul National University (Korea, Republic of)

    2016-08-26

    Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and {sup 64}Cu. HT29 (hTERT+) and U2OS (hTERT−) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus. In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo. - Highlights: • We developed new probes for imaging hTERT using Tat-conjugated IgM antibodies labeled with a fluorescent dye and radioisotope. • This probes could be used to overcome limitation of conventional antibody imaging system in live cell imaging. • This system could be applicable to monitor intracellular and intranuclear proteins in vitro and in vivo.

  6. Prevalence of human cytomegalovirus (HCMV antibodies among patients HIV/AIDS in Kurdistan province, 2015

    Directory of Open Access Journals (Sweden)

    Arezo Omati

    2016-12-01

    Full Text Available Background : Cytomegalovirus (CMV infection is one of the most common causes of death in people with immune deficiency diseases such as: organ or tissue recipients, patients with HIV/AIDS and newborn infants. The aim of this study was to determine the prevalence of anti-CMV antibodies in patients with HIV/AIDS in the Kurdistan province. Materials and Methods: This cross-sectional study carried out on 151 patients with HIV/AIDS covered by behavioral health counseling centers in Kurdistan province, 2015. For all patients the values of CMV antibodies, include IgG and IgM, were determined by ELISA technique and Diagnostic Kits (EIA WELL, Rome, Italy. Data analyses were done by use of t-test and multiple linear regression tests in stata software, version 13. Results: 116 (76.8% and 35 (23.2% of the patients were male and female, respectively. Mean (SD age of the patients were 39.3 (9.1 years. All studied patients were found positive for CMV-IgG (prevalence=100%, whereas only one case (0.7% was found positive for CMV-IgM. The relationship between age and CMV-IgG levels was statistically significant. Conclusion: The results showed that high prevalence of CMV in patients with HIV/AIDS, so in addition to paying more attention to the issue of HIV and CMV co-infection, about value of early antiretroviral treatment conducting further studies seems necessary.

  7. Insights into the biological features of the antigenic determinants recognized by four monoclonal antibodies in redia and adult stages of the liver fluke Fasciola hepatica.

    Science.gov (United States)

    Alba, Annia; Sánchez, Jorge; Hernández, Hilda; Mosqueda, Maryani; Rodríguez, Suanel Y; Capó, Virginia; Otero, Oscar; Alfonso, Carlos; Marcet, Ricardo; Sarracent, Jorge

    2016-09-01

    Fasciola hepatica is a digenean trematode which infects a wide variety of domestic animals and also humans. Previous studies have demonstrated that four monoclonal antibodies (Mabs) against the total extract of F. hepatica redia (named as 1E4, 6G11, 4E5 and 4G11) also recognized the excretion - secretion antigens (ES Ag) of adult parasites, which is a biologically-relevant mixture of molecules with functional roles during infection and immune evasion on definitive hosts. In the present report we describe the partial characterization of the epitopes recognized by these Mabs by heat treatment, mercaptoethanol reduction, pronase proteolysis and sodium peryodate oxidation, which suggested their predominant protein and conformational nature. Also, a comparative study using immunodetection assays on crude extracts and on histological sections of both rediae and adults of F. hepatica were performed to explore the expression pattern of the antigenic determinants in these developmental stages. From these experiments it was found that the Mabs reacted most likely with the same proteins of approximately 64 and 105 kDa present on both rediae and adult's extracts. However, the 1E4, 6G11 and 4E5 Mabs also recognized other molecules of the total extract of F. hepatica adults, a fact that constitutes an evidence of the antigenic variation between both stages and points at a certain biological relevance of the recognized antigenic determinants. Immunolocalization studies on histological sections revealed that all Mabs reacted with the tegument of F. hepatica in both rediae and adults stages, while the epitopes recognized by 1E4, 6G11 and 4E5 antibodies were also preferentially localized in the intestinal caeca and in different organs of the reproductive system of adult specimens. The immunogenicity of these antigenic determinants, their conserved status among different stages of the life cycle of F. hepatica and their presence in both tegument and ES Ag of adult parasites

  8. Validation of the kit ELISA (FAO, IAEA, PANAFTOSA) to determine antibodies against the virus of the foot-and-mouth disease

    International Nuclear Information System (INIS)

    Novel, Magaly; Ochoa, Carlos; Ramos, Pedro; Dominguez, Josefa de; Ruiz, Richard

    1997-01-01

    In Venezuela, endemic country for foot and mouth disease, the inmunoenzymatic test ELISA is not used in the routine diagnose for detection of antibodies against this illness. This fact determined that Panaftosa selected Venezuela as well as Argentina, Brazil, Colombia and Paraguay, to validate the kit developed by FAO - IAEA, in order to establish the point of cut value for each country. It is of high importance to precise the details of the technique in antigens (glycerinated or lyophilizated) so that, under equal conditions, national epidemic studies can be settle down, in such a way that its results can reorient the vaccination campaign and contribute to the Eradication of the Foot and Mouth Disease in Southamerican Countries. In this first stage 622 serums were processed, vaccinated animal (200), virgin (400), and coming from farms where the disease appeared (22). The analysis gave the screening in virgin animals among the 290 serums coming from Venezuela, with the result of 7 positive; this determined to continue insisting on requesting negative serums from Patagonia Argentina. The serum were received on September 14 of 1995 and mounting the screening in front of the Ag O and A, they were really negative to the ELISA test. From the screening of the 200 vaccinated animal serums, 48 were positive A (24%) and 33 positive to virus O (15.5); these results were taken with reservation, since the controls of antigens didn't give the value settled down in the protocol 1.0-1.5 [es

  9. Antibodies to Lactobacilli and Bifidobacteria in young children with different propensity to develop islet autoimmunity.

    Science.gov (United States)

    Talja, Ija; Kubo, Anna-Liisa; Veijola, Riitta; Knip, Mikael; Simell, Olli; Ilonen, Jorma; Vähä-Mäkilä, Mari; Sepp, Epp; Mikelsaar, Marika; Utt, Meeme; Uibo, Raivo

    2014-01-01

    The intestinal microbiota is essential to the maturation and homeostasis of the immune system. Immunoblot assays were used to establish the prevalence of serum IgG, IgM, and IgA antibodies specific for Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus rhamnosus GG proteins in young children presenting with or without type 1 diabetes (T1D). We demonstrated that children between the ages of 6 and 12 months had a substantial increase in the frequency of IgG antibodies specific for L. rhamnosus GG proteins. We measured IgG, IgM, and IgA class antibody reactivity against B. adolescentis DSM 20083, B. adolescentis DSM 20086, and B. longum DSM 20088 proteins demonstrating significantly higher IgA responses against B. adolescentis DSM 20083 strain proteins in children who developed islet autoimmunity and T1D later in life. B. adolescentis strains showed more IgM type antibodies in children who developed T1D later in life, but the difference was not statistically significant. B. longum proteins were recognized by IgG and IgA antibodies to a higher extent compared to other bacteria studied. These results confirm that differences in immune reactivity against some commensal strains in young children may represent a different risk factor for developing T1D.

  10. Antibodies to Lactobacilli and Bifidobacteria in Young Children with Different Propensity to Develop Islet Autoimmunity

    Directory of Open Access Journals (Sweden)

    Ija Talja

    2014-01-01

    Full Text Available The intestinal microbiota is essential to the maturation and homeostasis of the immune system. Immunoblot assays were used to establish the prevalence of serum IgG, IgM, and IgA antibodies specific for Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus rhamnosus GG proteins in young children presenting with or without type 1 diabetes (T1D. We demonstrated that children between the ages of 6 and 12 months had a substantial increase in the frequency of IgG antibodies specific for L. rhamnosus GG proteins. We measured IgG, IgM, and IgA class antibody reactivity against B. adolescentis DSM 20083, B. adolescentis DSM 20086, and B. longum DSM 20088 proteins demonstrating significantly higher IgA responses against B. adolescentis DSM 20083 strain proteins in children who developed islet autoimmunity and T1D later in life. B. adolescentis strains showed more IgM type antibodies in children who developed T1D later in life, but the difference was not statistically significant. B. longum proteins were recognized by IgG and IgA antibodies to a higher extent compared to other bacteria studied. These results confirm that differences in immune reactivity against some commensal strains in young children may represent a different risk factor for developing T1D.

  11. Diagnostic accuracy and comparison of two assays for Borrelia-specific IgG and IgM antibodies

    DEFF Research Database (Denmark)

    Dessau, Ram

    2013-01-01

    to represent the serological response of disseminated early Lyme borreliosis in general. Serum samples were obtained from 216 Danish blood donors as controls. By comparing sensitivity and specificity using pre-specified cut-off values, significant differences were found. However, using receiver operating...

  12. High Levels of IgA Antibodies to Helicobacter Pylori among Omani Women during Pregnancy and after Delivery.

    Science.gov (United States)

    Al-Balushi, Mohammed Said; Said, Elias Anthony; Hasson, Sidgi Syed; Al-Busaidi, Juma Zaid; Al-Reesi, Iman; Idris, Mohammed; Al-Ghafri, Wadha; Al-Kalbani, Moza; Al-Jabri, Ali Abdullah

    2016-06-01

    Helicobacter pylori (H. pylori), is a common infection in pregnant women accompanied by variations in the levels of the IgM, IgA and IgG antibody isotypes. The variations of anti-H. pylori antibodies during and after pregnancy, and the extent of protection they provide to the mother and the fetus are not completely understood. To investigate the changes of the anti-H. pylori IgM, IgA and IgG levels in healthy Omani pregnant women during pregnancy and 3 months after delivery. Serum samples obtained from 70 Omani healthy pregnant women, with no history of autoimmune diseases, were tested for anti-H. pylori IgM, IgA and IgG in the first trimester of pregnancy and 3 months after delivery. In parallel and as a control group, sera obtained from a group of 70 healthy non-pregnant Omani women were tested. The levels of anti-H. pylori IgM, IgA and IgG were measured using standard Enzyme Linked Immunosorbent Assays (ELISAs). Anti-H. pylori IgA levels were found to be significantly higher during pregnancy (p=0.046) and after delivery (p=0.02) when compared to the control group. Moreover, a significant increase in the levels of anti-H. pylori IgM, IgA and IgG was detected after delivery (p=0.002) when compared to the levels during pregnancy. Pregnancy is associated with an increase in the levels of anti-H. pylori IgA antibodies. In addition, anti-H. pylori IgM, IgG and IgA antibody levels increase after delivery.

  13. Theranostics Using Antibodies and Antibody-Related Therapeutics

    NARCIS (Netherlands)

    Moek, Kirsten L; Giesen, Danique; Kok, Iris C; de Groot, Derk Jan A; Jalving, Mathilde; Fehrmann, Rudolf S N; Lub-de Hooge, Marjolijn N; Brouwers, Adrienne H; de Vries, Elisabeth G E

    In theranostics, radiolabeled compounds are used to determine a treatment strategy by combining therapeutics and diagnostics in the same agent. Monoclonal antibodies (mAbs) and antibody-related therapeutics represent a rapidly expanding group of cancer medicines. Theranostic approaches using these

  14. IgM ELISA for leptospirosis diagnosis: a systematic review and meta-analysis

    Directory of Open Access Journals (Sweden)

    Maria Ines Rosa

    Full Text Available Abstract A systematic review with meta-analysis was performed to estimate the accuracy of IgM ELISA for Leptospirosis diagnosis. A search of Medline, Lilacs, Embase, Cochrane Central Register of Controlled Trials and Grey literature (Google Scholar and British Library was conducted. The medical subject headings (MeSHs and the words “leptospirosis”, “human leptospirosis” and “IgM ELISA” were used. Fifty-two studies were analyzed, which included 10,775 samples. The pooled sensitivity of all the studies was 86% (CI 95%, 85%-87% and specificity was 90% (CI 95%, 89%-91%. In studies of the acute phase, the sensitivity and specificity were 84% (CI 95%, 82%-85% and 91% (CI 95%, 90%-91%, respectively. In conclusion, IgM ELISA is sensitive for use as an initial screen for leptospiral infections.

  15. Type-specific IgG and IgA antibodies in old lymphogranuloma venerum determined by solid-phase radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Meurman, O.; Terho, P.; Sonck, C.E.

    1981-01-01

    A solid-phase radioimmunoassay (RIA) using egg-grown purified Chlamydia trachomatis lymphogranuloma venereum (LGV) serotypes L1, L2, and L3 as antigen was used to measure type-specific IgG and IgA antibodies in sera of 36 patients who had contracted LGV infection about 40 years ago. The RIA test gave compatible results with the standard microimmunofluorescence test, and by RIA it was possible to identify the infecting serotype in 30 out of 36 patients studied. In 28 cases this was L2 and in two cases L1. Each patient had IgG antibodies and most of them (80%) IgA antibodies to at least one of the LGV serotypes. The antibody titers were still high 40 years after the acute infection, being higher than in male patients with a recent chlamydial urethritis. Highest antibody titers were detected in LGV patients who had a severe disease with intestinal involvement.

  16. Homeostatic roles of naturally occurring antibodies: an overview.

    Science.gov (United States)

    Lutz, Hans U

    2007-12-01

    Immunoglobulins may have been developed in evolution to provide specificity for clearing body waste in the first animals with three germ layers. Tissue homeostasis in vertebrates comprises clearance of proteins released from lysed cells, elimination of altered plasma proteins, of senescent and apoptotic cells. Rather specific IgM and IgG naturally occurring antibodies (NAbs) to cytoplasmic and cytoskeletal proteins bind to proteins released from lysing cells and the IgG NAbs are slightly upregulated upon demand. Some of these NAbs along with complement have devastating effects when massive amounts of intracellular proteins are released during an infarct or an ischemia/reperfusion experiment. IgM NAbs to neoepitopes on plasma proteins/lipids help clear denatured proteins and are protective. IgG NAbs to an exposed protein, band 3 from red blood cells, bind to oligomerized band 3 and due to an affinity for C3 within their framework preferentially form C3b2-IgG complexes from nascent C3b. Thus, anti-band 3 NAbs gain potency by using avidity and generating a potent precursor of the amplifying C3 convertase. IgM NAbs to neoepitopes, which are generated by oxidized lipids forming Schiff bases with proteins, are protective and help clear this waste in atherosclerosis, but IgG antibodies (NAbs?) of the same specificity promote disease.

  17. A single point in protein trafficking by Plasmodium falciparum determines the expression of major antigens on the surface of infected erythrocytes targeted by human antibodies.

    Science.gov (United States)

    Chan, Jo-Anne; Howell, Katherine B; Langer, Christine; Maier, Alexander G; Hasang, Wina; Rogerson, Stephen J; Petter, Michaela; Chesson, Joanne; Stanisic, Danielle I; Duffy, Michael F; Cooke, Brian M; Siba, Peter M; Mueller, Ivo; Bull, Peter C; Marsh, Kevin; Fowkes, Freya J I; Beeson, James G

    2016-11-01

    Antibodies to blood-stage antigens of Plasmodium falciparum play a pivotal role in human immunity to malaria. During parasite development, multiple proteins are trafficked from the intracellular parasite to the surface of P. falciparum-infected erythrocytes (IEs). However, the relative importance of different proteins as targets of acquired antibodies, and key pathways involved in trafficking major antigens remain to be clearly defined. We quantified antibodies to surface antigens among children, adults, and pregnant women from different malaria-exposed regions. We quantified the importance of antigens as antibody targets using genetically engineered P. falciparum with modified surface antigen expression. Genetic deletion of the trafficking protein skeleton-binding protein-1 (SBP1), which is involved in trafficking the surface antigen PfEMP1, led to a dramatic reduction in antibody recognition of IEs and the ability of human antibodies to promote opsonic phagocytosis of IEs, a key mechanism of parasite clearance. The great majority of antibody epitopes on the IE surface were SBP1-dependent. This was demonstrated using parasite isolates with different genetic or phenotypic backgrounds, and among antibodies from children, adults, and pregnant women in different populations. Comparisons of antibody reactivity to parasite isolates with SBP1 deletion or inhibited PfEMP1 expression suggest that PfEMP1 is the dominant target of acquired human antibodies, and that other P. falciparum IE surface proteins are minor targets. These results establish SBP1 as part of a critical pathway for the trafficking of major surface antigens targeted by human immunity, and have key implications for vaccine development, and quantifying immunity in populations.

  18. Immunodeficiency in the chicken. II. Production of monomeric IgM following testosterone treatment or infection with Gumboro disease.

    Science.gov (United States)

    Ivanyi, J

    1975-06-01

    Chickens were treated at an early embryonal age with testosterone propionate or infected neonatally with the virus producing Gumboro disease. Their sera were subsequently fractionated by Sephadex G-200 chromatography, and showed a complete deficiency of IgG and the presence of IgM which was eluted with the 7S protein fraction. Purified and 125-I-labelled monomeric IgM was examined by SDS-acrylamide gel electrophoresis and found to contain both mu and light chains, together with a chain of intermediary size, which was absent from the patterns of 19S IgM or IgG.

  19. Prevalence and clinical associations of lupus anticoagulant, anticardiolipin antibodies, and anti-beta2-glycoprotein I antibodies in patients with systemic lupus erythematosus.

    Science.gov (United States)

    Woo, Kwang-Sook; Kim, Kyung-Eun; Kim, Jeong Man; Han, Jin-Yeong; Chung, Won-Tae; Kim, Kyeong-Hee

    2010-02-01

    The presence of antiphospholipid antibodies (aPLs) is associated with the clinical features of antiphospholipid syndrome (APS), which comprises venous and arterial thrombosis and pregnancy loss, and systemic lupus erythematosus (SLE). The prevalence of aPLs has been reported to be different in patient populations affected by either of these conditions. We performed a retrospective study to evaluate the prevalence and clinical associations of aPLs, including lupus anticoagulant (LAC), anticardiolipin (aCL), and anti-beta2-glycoprotein I antibodies (anti-beta2-GPI) in a cohort of Korean patients with SLE. This study included samples from 88 SLE patients for whom aPL testing had been advised between June 2006 and July 2009 at the Dong-A University Hospital. Serum and plasma samples were tested for LAC, aCL (IgG, IgM), and anti-beta2-GPI (IgG, IgM) antibodies. Clinical data from patients were obtained from a review of medical records. LAC was the most common (34.1% of total patients, 30/88) antibody, followed by IgM aCL (31.8%, 28/88), IgG aCL (18.2%, 16/88), and IgM and IgG anti-beta2-GPI (both 5.7%, 5/88 each). Positivity for LAC was strongly associated with venous/arterial thrombosis (P=0.002). LAC was the most common antibody detected in Korean SLE patients and is shown to have a significant association with the presence of venous/arterial thrombosis. The measurement of LAC may be clinically useful in identifying patients with SLE who are at a high risk for venous/arterial thrombosis.

  20. Tabhu: tools for antibody humanization

    DEFF Research Database (Denmark)

    Olimpieri, Pier Paolo; Marcatili, Paolo; Tramontano, Anna

    2015-01-01

    and time-consuming experiments. Here we present tools for antibody humanization (Tabhu) a web server for antibody humanization. Tabhu includes tools for human template selection, grafting, back-mutation evaluation, antibody modelling and structural analysis, helping the user in all the critical steps...... elicit unwanted and dangerous immunogenic responses. Antibody humanization methods are designed to produce molecules with a better safety profile still maintaining their ability to bind the antigen. This can be accomplished by grafting the non-human regions determining the antigen specificity...... of the humanization experiment protocol....

  1. [Clinical and immunological profile of 15 Moroccan patients with Hyper IgM syndrome].

    Science.gov (United States)

    Ouair, Hind; Benhsaien, Ibtihal; Jeddane, Leila; El Bakkouri, Jalila; Elhafidi, Naima; Rada, Noureddine; Najib, Jilali; Ailal, Fatima; Alj, Hanane Salih; Bousfiha, Ahmed Aziz

    2017-01-01

    Hyper IgM syndrome is a well known genetic (primary) immunodeficiency disorder which was first described in 1961. It is caused by B lymphocyte deficiency characterized by normal or elevated serum IgM levels and low or zero levels of IgG, IgA, IgE resulting from isotype-switching deficiency. Clinical manifestations are dominated by recurrent infections, especially involving the digestive tube of the ENT sphere and the lungs. This syndrome is caused by B-cell immunoglobulin class switch deficiency and decreased capacity to induce proliferation of T lymphocytes. The net result of these deficiencies is reflected in increased susceptibility to Pneumocystis jiroveci, Cryptosporidium spp and other intracellular organisms as well as high rate of bacterial and viral infections. This study aimed to illustrate the importance of understanding the pathophysiological mechanisms associated with this increased susceptibility to infections in order to allow a better diagnosis and therapy in patients with Hyper IgM syndrome (HIM).

  2. Snakes antibodies.

    Science.gov (United States)

    Gambón-Deza, Francisco; Sánchez-Espinel, Christian; Mirete-Bachiller, Serafín; Magadán-Mompó, Susana

    2012-09-01

    Immunoglobulins are basic molecules of the immune system of vertebrates. In previous studies we described the immunoglobulins found in two squamata reptiles, Anolis carolinensis and Eublepharis macularius. Snakes are squamata reptiles too but they have undergone an extreme evolutionary process. We therefore wanted to know how these changes affected their immunoglobulin coding genes. To perform this analysis we studied five snake transcriptomes and two genome draft sequences. Sequences coding for immunoglobulin M (IgM), immunoglobulin D (IgD) and two classes of immunoglobulin Y (IgY - named IgYa and IgYb-) were found in all of them. Moreover, the Thamnophis elegans transcriptome and Python molurus genome draft sequences showed a third class of IgY, the IgYc, whose constant region only presents three domains and lacks the CH2. All data suggest that the IgYb is the evolutionary origin of this IgYc. An exhaustive search of the light chains were carried out, being lambda the only light chain found in snakes. The results provide a clear picture of the immunoglobulins present in the suborder Serpentes. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. An enzyme-linked immunosorbent assay for the detection of mouse polyomavirus-specific antibodies in laboratory mice.

    NARCIS (Netherlands)

    H.W.J. Broeders; J. Groen (Jan); A.D.M.E. Osterhaus (Albert); G. van Steenis (Bert)

    1994-01-01

    textabstractAn enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantification of IgM and IgG serum antibodies to mouse polyomavirus (MPV). To evaluate the potential of this ELISA for the screening of laboratory rodents, serum samples from specific pathogen free (SPF)

  4. Antibody reactivities to glutamate-rich peptides of Plasmodium falciparum parasites in humans from areas of different malaria endemicity

    DEFF Research Database (Denmark)

    Jakobsen, P H; Theander, T G; Hviid, L

    1996-01-01

    Synthetic P. falciparum peptides were evaluated as tools in epidemiological investigations of malaria. Plasma IgM and IgG antibody reactivities against synthetic peptides covering sequences of glutamate-rich protein (GLURP) and acidic-basic repeat antigen (ABRA) were measured by ELISA...

  5. Antibody-mediated enhancement of human immunodeficiency virus type 1 infectivity is determined by the structure of gp120 and depends on modulation of the gp120-CCR5 interaction

    NARCIS (Netherlands)

    C. Guillon (Christophe); M. Schutten (Martin); P.H.M. Boers (Patrick); R.A. Gruters (Rob); A.D.M.E. Osterhaus (Albert)

    2002-01-01

    textabstractIn this study, we characterized the viral determinants of coreceptor usage in relation to susceptibility to antibody-mediated neutralization or enhancement of infectivity by using chimeras of three highly related human immunodeficiency virus type 1 (HIV-1) isolates of different

  6. Interferon gamma modulation of disease manifestation and the local antibody response to alphavirus encephalomyelitis.

    Science.gov (United States)

    Baxter, Victoria K; Griffin, Diane E

    2016-11-01

    Infection of mice with Sindbis virus (SINV) produces encephalomyelitis and provides a model for examination of the central nervous system (CNS) immune response to alphavirus infection. Clearance of infectious virus is accomplished through a cooperative effort between SINV-specific antibody and IFN-γ, but the regulatory interactions are poorly understood. To determine the effects of IFN-γ on clinical disease and the antiviral immune response, C57BL/6 mice lacking IFN-γ (Ifng-/-) or IFN-γ receptor (Ifngr1-/-) were studied in comparison to WT mice. Maximum production of Ifng mRNA and IFN-γ protein in the CNS of WT and Ifngr1-/- mice occurred 5-7 days after infection, with higher levels of IFN-γ in Ifngr1-/- mice. Onset of clinical disease was earlier in mice with impaired IFN-γ signalling, although Ifngr1-/- mice recovered more rapidly. Ifng-/- and Ifngr1-/- mice maintained body weight better than WT mice, associated with better food intake and lower brain levels of inflammatory cytokines. Clearance of infectious virus from the spinal cords was slower, and CNS, but not serum, levels of SINV-specific IgM, IgG2a and IgG2b were lower in Ifngr1-/- and Ifng-/- mice compared to WT mice. Decreased CNS antiviral antibody was associated with lower expression of mRNAs for B-cell attracting chemokines CXCL9, CXCL10 and CXCL13 and fewer B cells in the CNS. Therefore, IFN-γ signalling increases levels of CNS pro-inflammatory cytokines, leading to clinical disease, but synergistically clears virus with SINV-specific antibody at least in part by increasing chemokine production important for infiltration of antibody-secreting B cells into the CNS.

  7. Human anti-animal antibody interferences in immunological assays.

    Science.gov (United States)

    Kricka, L J

    1999-07-01

    The scope and significance of human anti-animal antibody interference in immunological assays is reviewed with an emphasis on human anti-animal immunoglobulins, particularly human anti-mouse antibodies (HAMAs). Anti-animal antibodies (IgG, IgA, IgM, IgE class, anti-isotype, and anti-idiotype specificity) arise as a result of iatrogenic and noniatrogenic causes and include human anti-mouse, -rabbit, -goat, -sheep, -cow, -pig, -rat, and -horse antibodies and antibodies with mixed specificity. Circulating antibodies can reach gram per liter concentrations and may persist for years. Prevalence estimates for anti-animal antibodies in the general population vary widely and range from HAMA, which causes both positive and negative interferences in two-site mouse monoclonal antibody-based assays. Strategies to prevent the development of human anti-animal antibody responses include immunosuppressant therapy and the use of humanized, polyethylene glycolylated, or Fab fragments of antibody agents. Sample pretreatment or assay redesign can eliminate immunoassay interferences caused by anti-animal antibodies. Enzyme immunoassays, immunoradiometric assays, immunofluorescence, and HPLC assays have been designed to detect HAMA and other anti-animal antibodies, but intermethod comparability is complicated by differences in assay specificity and lack of standardization. Human anti-animal antibodies often go unnoticed, to the detriment of patient care. A heightened awareness on the part of laboratory staff and clinicians of the problems caused by this type of interference in routine immunoassay tests is desirable. Efforts should be directed at improving methods for identifying and eliminating this type of analytical interference.

  8. Antibodies Targeting EMT

    Science.gov (United States)

    2017-10-01

    determine their targets on the cell. The newly discovered antibodies will then be engineered for utility as new highly specific drugs and diagnostics in...are from the aldo-keto reductase family (AKRs). Remarkably, 3 of the top 10 genes with induction in the mesenchymal TES2b cells Figure 1. Amino

  9. Seroprevalence of parvovirus B19 antibodies and evidence of viremia among Nigerian patients with sickle cell anemia

    Science.gov (United States)

    Iwalokun, Bamidele Abiodun; Iwalokun, Senapon Olusola; Hodonu, Semande Olufunmilayo

    2013-01-01

    Clinical, biochemical and molecular evidence for the sickle cell anemia (SCA) crisis in Nigerian patients arising from parvovirus b19 infection remains inadequate. This study determined the prevalence and correlates of anti-parvovirus b19 antibodies in a population of SCA patients and non-SCA healthy controls in Lagos, Nigeria. In this prospective cross-sectional study, we enrolled 73 confirmed SCA patients from 5 district hospitals in Lagos and 81 sex and age-matched non-SCA healthy controls. Serum sample from each study participant was screened for anti-parvovirus b19 by ELISA and PCR techniques. Standard biomedical assays were also done. Anti-parvovirus b19 IgM and IgG antibodies were detected in 22 (14.3%) and 97 (62.9%) of the 154 sera screened, 13 (17.8%) and 45 (61.6%) in SCA patients; 9 (11.1%) and 52 (64.2%) in non-SCA controls. The overall seronegativity rate was 19.5%. Parvovirus B19 DNA was found in 2 (11.1%) of the 18 IgM seropositive SCA serum samples screened. On the whole, parvovirus b19 infection was more commonly asymptomatic in non-SCA controls but caused significant elevation in liver enzymes in infected SCA patients (P parvovirus b19 infection increased 65 times during unsteady state among the SCA patients. Although no deaths of infected patients were recorded during the study, age below 12 years, hospitalization and overcrowded environment were risk factors for infection. We conclude that parvovirus b19 is common in SCA patients, incurring greater susceptibility to infections. PMID:23885266

  10. Moisture content determination in an antibody-drug conjugate freeze-dried medicine by near-infrared spectroscopy: A case study for release testing.

    Science.gov (United States)

    Clavaud, Matthieu; Roggo, Yves; Dégardin, Klara; Sacré, Pierre-Yves; Hubert, Philippe; Ziemons, Eric

    2016-11-30

    The use of Near-infrared spectroscopy (NIRS) as a fast and non-destructive technique was employed for moisture content (MC) determination in Antibody-drug conjugates (ADCs) in replacement to Karl Fischer (KF) method. The lab analysis of ADCs, high potent medicines, should be performed in conditions ensuring the operator's safety and using secured analytical tools like NIRS. A NIRS method was first developed and validated in compliance with current guidelines. The novelty of this work first lies in the large number of samples prepared for a wide moisture calibration range of 0.51%-4.01%. Then, the classical Partial Least Square (PLS) regression was used as chemometric tool for the computation of the model. Excellent predictive calibration results were shown. A coefficient of correlation (r) value of 0.99 was obtained. An intercept value of 0.02 and a slope of 0.99 were observed, while the root mean square error of calibration (RMSEC) and the root mean square error of prediction (RMSEP) were respectively 0.10% and 0.12%. In addition, instrumentation, model performances and robustness of the method were evaluated, demonstrating the validation results. Calibration transfer issue and impact of the number of samples were also evaluated. Consequently, a validation strategy was introduced as a basis for submission to the health authorities' for release and stability activities in a cGMP environment in replacement of the KF method. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Anti-N antibody reacting at 37°C: An unusual occurrence interfering with routine testing: Two interesting cases

    Directory of Open Access Journals (Sweden)

    Vijay Kumawat

    2015-01-01

    Full Text Available Most anti-N antibodies are naturally occurring, IgM antibodies, and not active above 25°C and are not clinically significant but IgG anti- N has also been described. Immune anti-N resulting from multiple transfusions does occur & has been implicated as the cause of hemolytic transfusion reactions and mild hemolytic disease of fetus and newborn. Anti- N reacting at room temperature can be a cause for ABO blood group discrepancy

  12. Development of a highly specific and sensitive rubella immunoglobulin M antibody capture enzyme immunoassay that uses enzyme-labeled antigen.

    OpenAIRE

    Seppänen, H

    1990-01-01

    An enzyme immunoassay (EIA) for serum immunoglobulin M (IgM) antibodies to rubella virus based on enzyme labeling of viral antigen was developed. The sensitivity of the EIA for the detection of recent rubella virus infection was evaluated by using 115 rubella-IgM-antibody-positive serum specimens, which were confirmed as positive by Rubazyme M (Abbott Diagnostics). In addition, 12 individuals, 2 of whom were exposed to rubella through vaccination and 10 of whom were exposed through natural in...

  13. Novel enzyme immunoassay system for simultaneous detection of six subclasses of antiphospholipid antibodies for differential diagnosis of antiphospholipid syndrome.

    Science.gov (United States)

    Nojima, Junzo; Motoki, Yukari; Hara, Kazusa; Sakata, Toshiyuki; Ichihara, Kiyoshi

    2017-06-01

    : Antiphospholipid syndrome, which often complicates systemic lupus erythematosus (SLE), features high occurrence of arterial and/or venous thrombosis and recurrent fetal loss. However, which antibody subclass contributes to which clinical event remains uncertain. We newly developed an up-to-date enzyme immunoassay system using the AcuStar automated analyzer (Instrumentation Laboratory, Bedford, Massachusetts, USA) for parallel detection of six subclasses of antiphospholipid antibodies (aPLs): anticardiolipin antibodies (aCL) of IgG, IgM, and IgA and anti-β2-glycoprotein I antibodies (aβ2GPI) of IgG, IgM, and IgA. They were measured in 276 healthy volunteers and 138 patients with SLE: 45 with thromboembolic complications (29 arterial; 16 venous) and 93 without. Lupus anticoagulant activity in their plasma was measured according to the guidelines recommended by the Subcommittee on Lupus Anticoagulant/Phospholipid-Dependent Antibodies. aCL/β2GPI was measured with a standard ELISA kit commonly used in Japan. The positive results of IgG aCL, IgA aCL, and IgG aβ2GPI were closely associated with thromboembolic complications, whereas IgM aCL and IgM aβ2GPI were not. receiver operating characteristic analysis revealed that the accuracy of predicting thromboembolic complications based on the composite test results of the former three antibodies were obviously higher than by each alone. Regarding agreement with the test results of lupus anticoagulant activity, IgG aβ2GPI showed the closest match. Patients with SLE frequently possess various combinations of the six aPL subclasses, and this antibody spectrum is closely associated with thromboembolic events in these patients. This new automated enzyme immunoassay system allows simultaneous analysis of the profile of aPL subclasses for the differential diagnosis of antiphospholipid antibody syndrome in its early stage.

  14. Antibody biotechnology

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-06

    Jul 6, 2009 ... and automated, the hybrid cells can be stored for many years in liquid nitrogen and antibodies production is homogeneous. The hybridoma method .... they may be modified to vehicle active molecules such as radio-isotopes, toxins, cytokines, enzyme etc. In these cases, the therapeutic effect is due to ...

  15. Catalytic Antibodies

    Indian Academy of Sciences (India)

    The ability of the highly evolved machinery of immune system to produce structurally and functionally complex ... to Pauling, if the structure of the antigen binding site of antibodies were to be produced in a random ..... where the immune system of the body is destructive, as in autoimmune disorders or after organ transplant.

  16. Catalytic Antibodies

    Indian Academy of Sciences (India)

    While chemistry provides the framework for understanding the structure and function of biomolecules, the immune sys- tem provides a highly evolved natural process to generate one class of complex biomolecules – the antibodies. A combination of the two could be exploited to generate new classes of molecules with novel ...

  17. [Significance of anti-citrullinated human papilloma virus-47 E2(345-362) peptide antibodies in diagnosis of early-stage rheumatoid arthritis].

    Science.gov (United States)

    Li, Gui-ye; Sun, Xiao-lin; Li, Yun; Jia, Ru-lin; Jia, Yuan; Li, Zhan-guo

    2013-06-18

    To investigate the prevalence and the diagnostic values of antibodies to the citrullinated HPVP in early-stage rheumatoid arthritis. Antibodies against HPVP and citrullinated HPVP were detected by ELISA in the sera of 101 patients with early-stage RA, 149 patients with other rheumatic diseases and 40 healthy controls. The prevalence and diagnostic values of these antibodies for early-stage RA were analyzed by statistical software. (1)The prevalence of IgG, IgM antibodies to citrullinated HPVP in early-stage RA were significantly higher than that in the patients with other rheumatic diseases as well as in the healthy individuals.(2)The diagnostic sensitivity of IgG and IgM citrullinated HPVP antibodies in early-stage RA were 62.4% and 66.3% respectively and the specificity value of the two antibody isotypes were 88.7% and 89.6%,similar to that of the anti-CCP antibody. (3)The positivity rates of IgG and IgM antibodies against citrullinated HPVP were 59.4% and 71.9% in IgM-RF negative early-stage RA patients, and 39.4% and 51.5% in anti-CCP antibody negative early-stage RA patients. (4) The DAS28 score [ IgG (6.3±1.0) vs. (5.6±0.9), P=0.002; IgM (6.2±1.1) vs. (5.6±0.8), P=0.008] , X-ray stages (IgG 56.1% vs. 21.2%, P=0.001;IgM 50.9% vs. 29.4%, P=0.036),anti-CCP antibodies(IgG 96.8% vs. 55.3%, P=0.001; IgM 89.6% vs. 64.7%, P=0.023) in citrullinated HPVPP positive patients were higher than those of citrullinated HPVP negative patients. Additionally, the levels of ESR[IgG(70.3±32.4)vs.(51.9±27.8), P=0.004; IgM (67.4±31.5)vs.(53.8±27.7), P=0.035] in citrullinated HPVP positive patients were higher than those of negative patients (Pbiomarkers for early-stage RA diagnosis, and are related to disease activity and joint damage.

  18. Cytokine measurements and possible interference from heterophilic antibodies--problems and solutions experienced with rheumatoid factor

    DEFF Research Database (Denmark)

    Bartels, Else Marie; Ribel-Madsen, Søren

    2013-01-01

    these. Some antibodies, such as heterophilic and human anti-animal antibodies, are able to interfere with all immunoassays, but the immunometric techniques are most prone to serious interference from this source. Another type, rheumatoid factor (RF) is a composite of different autoimmune antibodies...... which can be present in both blood and synovial fluid. RF is present in some arthritic diseases as well as in some other medical conditions. When present, especially RF IgM is known to interfere with the immunometric measurements. A possible and affordable solution to diminish this interference is PEG...

  19. igm Robots with laser measurement for an economic repair welding of equipment components of open-pit minings; igm-Roboter mit Laservermessung zum wirtschaftlichen Reparaturschweissen von Tagebauausruestungsteilen

    Energy Technology Data Exchange (ETDEWEB)

    Wihsbeck, Martin [igm Robotersysteme AG, Wiener Neudorf (Austria); Wache, Wolfgang [Vattenfall Europe Mining AG, Spremberg (Germany). Schwarze Pumpe; Springfeld, Peter

    2012-08-15

    In the Lusatian area,Vattenfall Europe AG prosecutes four open-pit minings whose lignite directly is supplied to power stations. A quarter of Germany's total electricity supply is based on lignite. The lignite mining in the area of the Lusatian open-pit minings is particularly efficient due to the horizontal orientation of the Lusatian coal seams. The movable components of the overburden conveyor bridges as well as the bucket wheel excavators and bucket ladder dredgers are subject to a tremendous wear. It is not economically feasible to refurbish certain wear parts. Such parts include Turas corners. In order to refurbish such Turas corners nevertheless, the Austrian company igm robot system AG (Wiener Neudorf, Austria) offers a welding robot.

  20. IgM AUTOANTIBODIES TO DNA IN BLOOD SERUM OF THE PATIENTS WITH HEMORRHAGIC FEVER WITH RENAL SYNDROME

    Directory of Open Access Journals (Sweden)

    D. G. Ishmukhametova

    2010-01-01

    Full Text Available Levels of IgM autoantibodies (AAbs to native (double-stranded and denaturated (single-stranded DNA were studied in blood serum of sixty patients with hemorrhagic fever with renal syndrome (HFRS and twenty-five healthy persons, using an ELISA technique. The median levels of IgM AAbs to double-stranded DNA in blood serum of healthy persons and HFRS patients corresponded to 0.41 and 0.53 arbitrary units, respectively. Thus, the difference between the samples from HFRS and healthy persons proved to be non-significant. The median level of IgM AAbs to single-stranded DNA in blood sera of HFRS patients (0.71 arbitrary units did significantly exceed serum values of healthy persons (0.57 arbitrary units. A probable involvement of IgM AAbs into regulation of IgG AAbs' production during virus-induced activation of autoimmune events in HFRS patients is discussed.

  1. Humoral and secretory antibodies to Ureaplasma diversum in heifers following subcutaneous vaccination and vaginal infection.

    Science.gov (United States)

    Mulira, G L; Saunders, J R

    1994-04-01

    We measured antibody levels in serum and cervicovaginal mucus (CVM) of four heifers vaccinated with two inoculations of killed Ureaplasma diversum strain 2312 in incomplete Freund's adjuvant (IFA) two weeks apart, and six heifers given a placebo. Two weeks later, the vaccinates and four placebo heifers, were challenged by intravaginal inoculation with 6.4 x 10(8) colony-forming units of the homologous U. diversum strain. The remaining two placebo heifers served as unvaccinated, unchallenged controls. Antibody levels in serum and CVM of all heifers were determined by an enzyme-linked immunosorbent assay (ELISA). Vaccination stimulated specific IgG1 and IgG2 responses in serum and CVM but only a slight IgM and no IgA response. In both vaccinate and placebo heifers, subsequent intravaginal challenge resulted in a granular vulvitis (GV) with a predominant IgA response in the CVM. The GV gradually subsided during the 35 day observation period but ureaplasmas were consistently demonstrated by culture. We concluded that subcutaneous vaccination stimulated a specific, albeit nonprotective, IgG response in serum and CVM. In contrast, vaginal infection primarily induced a mucosal IgA response.

  2. Hantavirus Pulmonary Syndrome in Santa Cruz, Bolivia: Outbreak Investigation and Antibody Prevalence Study

    Science.gov (United States)

    Montgomery, Joel M.; Blair, Patrick J.; Carroll, Darin S.; Mills, James N.; Gianella, Alberto; Iihoshi, Naomi; Briggiler, Ana M.; Felices, Vidal; Salazar, Milagros; Olson, James G.; Glabman, Raisa A.; Bausch, Daniel G.

    2012-01-01

    We report the results of an investigation of a small outbreak of hantavirus pulmonary syndrome in 2002 in the Department of Santa Cruz, Bolivia, where the disease had not previously been reported. Two cases were initially reported. The first case was a physician infected with Laguna Negra virus during a weekend visit to his ranch. Four other persons living on the ranch were IgM antibody-positive, two of whom were symptomatic for mild hantavirus pulmonary syndrome. The second case was a migrant sugarcane worker. Although no sample remained to determine the specific infecting hantavirus, a virus 90% homologous with Río Mamoré virus was previously found in small-eared pygmy rice rats (Oligoryzomys microtis) trapped in the area. An antibody prevalence study conducted in the region as part of the outbreak investigation showed 45 (9.1%) of 494 persons to be IgG positive, illustrating that hantavirus infection is common in Santa Cruz Department. Precipitation in the months preceding the outbreak was particularly heavy in comparison to other years, suggesting a possible climatic or ecological influence on rodent populations and risk of hantavirus transmission to humans. Hantavirus infection appears to be common in the Santa Cruz Department, but more comprehensive surveillance and field studies are needed to fully understand the epidemiology and risk to humans. PMID:23094116

  3. Cocoa Diet and Antibody Immune Response in Preclinical Studies

    Directory of Open Access Journals (Sweden)

    Mariona Camps-Bossacoma

    2017-06-01

    Full Text Available The ability of cocoa to interact with the immune system in vitro and in vivo has been described. In the latter context, a cocoa-enriched diet in healthy rats was able to modify the immune system’s functionality. This fact could be observed in the composition and functionality of lymphoid tissues, such as the thymus, spleen, and lymph nodes. Consequently, immune effector mechanisms, such as antibody synthesis, were modified. A cocoa-enriched diet in young rats was able to attenuate the serum levels of immunoglobulin (Ig G, IgM, and IgA and also the intestinal IgM and IgA secretion. Moreover, in immunized rats, the intake of cocoa decreased specific IgG1, IgG2a, IgG2c, and IgM concentrations in serum. This immune-regulator potential was then tested in disease models in which antibodies play a pathogenic role. A cocoa-enriched diet was able to partially prevent the synthesis of autoantibodies in a model of autoimmune arthritis in rats and was also able to protect against IgE and T helper 2-related antibody synthesis in two rat models of allergy. Likewise, a cocoa-enriched diet prevented an oral sensitization process in young rats. In this review, we will focus on the influence of cocoa on the acquired branch of the immune function. Therefore, we will focus on how a cocoa diet influences lymphocyte function both in the systemic and intestinal immune system. Likewise, its potential role in preventing some antibody-induced immune diseases is also included. Although further studies must characterize the particular cocoa components responsible for such effects and nutritional studies in humans need to be carried out, cocoa has potential as a nutraceutical agent in some hypersensitivity status.

  4. Isomerization and Oxidation in the Complementarity-Determining Regions of a Monoclonal Antibody: A Study of the Modification-Structure-Function Correlations by Hydrogen-Deuterium Exchange Mass Spectrometry.

    Science.gov (United States)

    Yan, Yuetian; Wei, Hui; Fu, Ya; Jusuf, Sutjano; Zeng, Ming; Ludwig, Richard; Krystek, Stanley R; Chen, Guodong; Tao, Li; Das, Tapan K

    2016-02-16

    Chemical modifications can potentially change monoclonal antibody's (mAb) local or global conformation and therefore impact their efficacy as therapeutic drugs. Modifications in the complementarity-determining regions (CDRs) are especially important because they can impair the binding affinity of an antibody for its target and therefore drug potency as a result. In order to understand the impact on mAb attributes induced by specific chemical modifications within the CDR, hydrogen-deuterium exchange mass spectrometry (HDX MS) was used to interrogate the conformational impact of Asp isomerization and Met oxidation in the CDRs of a model monoclonal antibody (mAb1). Our results indicate that despite their proximity to each other, Asp54 isomerization and Met56 oxidation in CDR2 in the heavy chain of mAb1 result in opposing conformational impacts on the local and nearby regions, leading directly to different alterations on antibody-antigen binding affinity. This study revealed direct evidence of local and global conformational changes caused by two of the most common degradation pathways in the CDRs of a mAb and identified correlations between chemical modification, structure, and function of the therapeutic monoclonal antibody.

  5. Radioimmunodetection of human melanoma tumor xenografts with human monoclonal antibodies

    International Nuclear Information System (INIS)

    Gomibuchi, Makoto; Saxton, R.E.; Lake, R.R.; Katano, Mitsuo; Irie, R.F.

    1986-01-01

    A human IgM monoclonal antibody has been established that defines a tumor-associated membrane antigen expressed on human melanoma cells. The antigen has been identified as the ganglioside GD2. In this paper, the authors describe the potential usefulness of the human monoclonal antibody for radioimaging. Nude mice bearing tumors derived from a human melanoma cell line were used as a model. Antibody activity was degradated significantly after labeling with 131 I by the use of a modified chloramine-T method. After testing various concentrations, labeled antibody of a specific activity of 2.8μCi/μg produced the best results. Balb/c nude mice bearing a GD2-positive M14 melanoma cell line were injected with 10-30μg of labeled antibody, and its radiolocalization in different organs and in the whole body were evaluated. The best tumor image was obtained on Day 6. The labeled antibody uptake ratio between tumor and muscle was 9.2:1; the ratio between tumor and liver was 1.4:1. These studies represent the first report of experimental tumor imaging with human monoclonal antibody. Human monoclonals will probably prove to be superior reagents for tumor imaging in melanoma patients if the problem of anti-body radiolysis is resolved. (author)

  6. Value of HAMA--determination in clinical practice--an overview.

    Science.gov (United States)

    Hasholzner, U; Stieber, P; Meier, W; Lamerz, R

    1997-01-01

    HAMAs (human anti mouse antibodies) in serum of patients may be stimulated as an immunologic reaction to the application of animal protein. They are differentiated into heterophilic (species-unspecific) and mouse-specific anti-isotypic (often IgG1) or-idiotypic antibodies as well as according to the human immunoglobulin type (IgG or IgM). Besides infrequent clinically immunologic side effects (anaphylactic reaction), their influence on tumor marker measurements predominates in the follow-up of tumor patients by the possible impairment of newly applied monoclonal antibodies. The detection of HAMAs is performed on suspicion of a tumor by self-made or commercial tests, however, there is no consensus standardization of tests, standards and controls. Experiences are reported herein on the occurrence of heterophilic antibodies, reasons of suspicion, detection and removal, comparative determination of HAMAs by two commercially available tests (ImmunoSTRIP HAMA, Enzygnost) and HAMA induction in patients following immunoscintigraphy (OC-125, n = 27). Besides the rare appearance of HAMAs in patients without pretreatment or following biologic therapy, i.v. application of monoclonal antibodies for diagnosis (immunoscintigraphy) or therapy represents the most frequent reason for HAMA development that should always be ruled out in case of anamnestic indication and clinically unexplained tumor marker changes.

  7. Anti-Toxoplasma antibodies in Egyptian rheumatoid arthritis patients.

    Science.gov (United States)

    El-Henawy, Abeer A; Hafez, Eman Abdel Razek; Nabih, Nairmen; Shalaby, Naglaa M; Mashaly, Mervat

    2017-05-01

    To assess seroprevalence of anti-Toxoplasma gondii antibodies; both IgG and IgM in Egyptian rheumatoid arthritis (RA) patients versus a non-RA group and to compare anti-Toxoplasma antibodies seroprevalence among RA patients receiving traditional treatment and RA patients treated with biologic drug. 60 RA patients and 60 healthy controls were enrolled in the study. Patients were categorized into two groups: one group included 30 patients receiving disease modifying anti-rheumatic drugs (DMARDs), while the other group included 30 patients receiving biologic agent, infliximab, a TNF-α antagonist. Serum samples of all investigated persons were examined for anti-Toxoplasma antibodies. RA activity markers including rheumatoid factor, anti-cyclic citrullinated protein antibodies, C reactive protein, ESR in addition to disease activity score 28 (DAS28) of RA patients were also evaluated to explore their association with Toxoplasma seropositivity. Anti-Toxoplasma IgG antibodies were detected among 46/60 RA patients (76.7%) versus 29/60 controls (48.3%), (p = 0.001). Anti-Toxoplasma IgG titre was higher among RA group [median, (range) = 232.940 (8.949-653.242) IU/ml] than among controls [median, (range) = 68.820 (2.450-318.945) IU/ml], (p Toxoplasma IgG antibodies. No positive anti-Toxoplasma IgM was detected. A positive correlation was detected between anti-Toxoplasma IgG titre and disease activity markers. Higher seroprevalence of anti-Toxoplasma IgG antibodies among RA patients compared to controls reflects an association between latent Toxoplasma infection and RA. Our findings support previous studies and necessitate future large-scale studies to elucidate the exact role of Toxoplasma whether a trigger of autoimmunity in RA or an effect of immunosuppression.

  8. Epstein-Barr virus early antigen diffuse (EBV-EA/D)-directed immunoglobulin A antibodies in systemic lupus erythematosus patients.

    Science.gov (United States)

    Draborg, A H; Jørgensen, J M; Müller, H; Nielsen, C T; Jacobsen, S; Iversen, L V; Theander, E; Nielsen, L P; Houen, G; Duus, K

    2012-08-01

    We sought to determine whether the serological response towards lytic cycle antigens of Epstein-Barr virus (EBV) is altered in systemic lupus erythematosus (SLE) patients. We used enzyme-linked immunosorbent assay (ELISA) to investigate the prevalence of EBV early antigen diffuse (EBV-EA/D) antibodies in sera from 60 patients with SLE, 40 with scleroderma (SSc), 20 with primary Sjögren's syndrome (pSS), 20 with rheumatoid arthritis (RA), 20 healthy controls, and also subjects with various circulating autoantibodies. Samples from patients were obtained from clinics specialized within the diseases in Denmark and Sweden and samples from healthy controls were obtained from volunteers. A significant elevated titre of immunoglobulin (Ig)A, IgG, and IgM EBV-EA/D antibodies was found in SLE patients compared to healthy controls, a finding not explained by immunosuppressive treatment or disease activity. The largest difference was observed for IgA EBV-EA/D antibodies (p = 0.0013) with a seropositive rate of 58% in SLE patients and 0% in healthy controls. RA and SSc patients and individuals seropositive for anti-Scl-70 were additionally found to have elevated titres of IgA EBV-EA/D antibodies (40%, p = 0.014; 60%, p = 0.015; and 38.5%, p = 0.045, respectively). However, the titres were generally lower than in SLE patients. Our findings support an association between EBV and SLE. The elevated titre of EBV-EA/D-directed IgA antibodies found in SLE patients could suggest reactivation of EBV in epithelial cells or reinfection of epithelial cells after reactivation in B cells, indicating lack of control of the latent infection.

  9. Nature of the bifunctional chelating agent used for radioimmunotherapy with yttrium-88 monoclonal antibodies: critical factors in determining in vivo survival and organ toxicity

    Energy Technology Data Exchange (ETDEWEB)

    Kozak, R.W.; Raubitschek, A.; Mirzadeh, S.; Brechbiel, M.W.; Junghaus, R.; Gansow, O.A.; Waldmann, T.A. (Center for Biologics Evaluation and Research, FDA, Bethesda, MD (USA))

    1989-05-15

    One factor that is critical to the potential effectiveness of radioimmunotherapy is the design of radiometal-chelated antibodies that will be stable in vivo. Stability in vivo depends on the condition that both the chelate linkage and radiolabeling procedures not alter antibody specificity and biodistribution. In addition, synthesis and selection of the chelating agent is critical for each radiometal in order to prevent inappropriate release of the radiometal in vivo. In the present study, we compare the in vivo stability of seven radioimmunoconjugates that use different polyaminocarboxylate chelating agents to complex yttrium-88 to the mouse anti-human interleukin-2 receptor monoclonal antibody, anti-Tac. Chelate linkage and radiolabeling procedures did not alter the immunospecificity of anti-Tac. In order to assess whether yttrium was inappropriately released from the chelate-coupled antibody in vivo, iodine-131-labeled and yttrium-88 chelate-coupled antibodies were simultaneously administered to the same animals to correlate the decline in yttrium and radioiodinated antibody activity. The four stable yttrium-88 chelate-coupled antibodies studied displayed similar iodine-131 and yttrium-88 activity, indicating minimal elution of yttrium-88 from the complex. In contrast, the unstable yttrium-88 chelate-coupled antibodies had serum yttrium-88 activities that declined much more rapidly than their iodine-131 activities, suggesting loss of the radiolabel yttrium-88 from the chelate. Furthermore, high rates of yttrium-88 elution correlated with deposition in bone. Four chelating agents emerged as promising immunotherapeutic reagents: isothiocyanate benzyl DTPA and its derivatives 1B3M, MX, and 1M3B.

  10. Enhanced jejunal production of antibodies to Klebsiella and other Enterobacteria in patients with ankylosing spondylitis and rheumatoid arthritis.

    Science.gov (United States)

    Mäki-Ikola, O; Hällgren, R; Kanerud, L; Feltelius, N; Knutsson, L; Granfors, K

    1997-07-01

    To measure gut immunity directly in jejunal fluid in patients with ankylosing spondylitis (AS) and rheumatoid arthritis (RA). Antibodies against three different Enterobacterias were measured in jejunal perfusion fluids (collected by a double balloon perfusion device) of 19 patients with AS, 14 patients with RA, and 22 healthy controls using enzyme linked immunosorbent assay. The AS patients had significantly increased jejunal fluid concentrations of IgM, IgG, and IgA class antibodies against Klebsiella pneumoniae, and IgM and IgA class antibodies against Escherichia coli and Proteus mirabilis compared with healthy controls. When compared with the patients with RA, the AS patients had higher concentrations of IgA and IgG class antibodies only against K pneumoniae. The RA patients had higher IgM class antibody concentrations against all three studied Enterobacterias, when compared with the healthy controls, suggesting an enhanced mucosal immune response in these patients. A three month treatment with sulphasalazine did not decrease enterobacterial antibody concentrations in the 10 patients with AS. There is strong direct evidence for an abnormal mucosal humoral immune response particularly to K pneumoniae in patients with AS.

  11. Comparison of different immunoassay methods to detect human anti-drug antibody using the WHO erythropoietin antibody reference panel for analytes.

    Science.gov (United States)

    Shibata, Hiroko; Nishimura, Kazuko; Miyama, Chizuru; Tada, Minoru; Suzuki, Takuo; Saito, Yoshiro; Ishii-Watabe, Akiko

    2018-01-01

    Development of an appropriate assay to detect anti-drug antibody (ADA) is important for assessing immunogenicity to therapeutic protein products. However, characterizing ADA assay methods is difficult because human ADA as a reference standard is not available in most cases. We compared the analytical performance of three ligand-binding assay methods for ADA, namely, surface plasmon resonance (SPR), electrochemiluminescence (ECL), and biolayer interferometry (BLI) methods, by using the anti-erythropoietin (EPO) monoclonal antibody reference panel developed by the World Health Organization (WHO) in 2015. Dose-dependent binding responses were observed for all nine anti-EPO antibodies in the anti-EPO panel by the SPR and BLI methods. In contrast, the ECL method did not clearly detect binding of low-affinity anti-EPO antibodies. Regarding IgG2 and IgM antibodies derived from the same clone, IgG2 exhibited a higher binding response in the SPR assay, whereas the IgM binding response was higher than that of IgG2 in the ECL assay. In the case of the BLI method, there was no consistent pattern observed in the binding responses of IgG2 or IgM. Results of the anti-EPO antibody reference panel, which contains a variety of monoclonal antibodies, indicated that the ability to detect ADAs differed among these assay methods. Therefore, with ligand-binding assays, differences in assay platforms can affect the sensitivity and other characteristics of assays to detect ADAs. These results show that understanding the analytical performance of ADA assays is important for an appropriate assessment of immunogenicity. Our study also indicated the benefits of using the established human ADA reference panel to assess the assay methods for ADA detection. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Experimental evidence for direct in situ binding of IgM and IgT to early trophonts of Ichthyophthirius multifiliis (Fouquet) in the gills of rainbow trout, Oncorhynchus mykiss (Walbaum)

    DEFF Research Database (Denmark)

    Jørgensen, Louise von Gersdorff; Heinecke, Rasmus Demuth; Skjødt, Karsten

    2011-01-01

    , Oncorhynchus mykiss, shortly (2 h) after invasion. No binding of IgT and no or only a weak binding of IgM was observed on the parasites in the gills of similarly exposed but naïve rainbow trout. This study indicates that antibodies play an important part in the protection of immune fish against Ich although......Freshwater fish are able to mount a protective immune response against the parasite Ichthyophthirius multifiliis (Ich) following a non-lethal exposure. Factors involved in immunity comprise cellular and humoral factors, but antibodies have been suggested to play a prominent role in protection...

  13. Antiphosphatidylserine/prothrombin antibodies as biomarkers to identify severe primary antiphospholipid syndrome.

    Science.gov (United States)

    Hoxha, Ariela; Mattia, Elena; Tonello, Marta; Grava, Chiara; Pengo, Vittorio; Ruffatti, Amelia

    2017-05-01

    Anti-phosphatidylserine/prothrombin (aPS/PT) antibodies have begun to be considered potentional biomarkers for antiphospholipid syndrome (APS). This cohort study investigate the role of aPS/PT antibodies as a risk factor for severe APS by evaluating the association between those antibodies and clinical/laboratory profiles of APS. Plasma/serum samples from 197 APS patients, 100 healthy subjects and 106 patients with autoimmune diseases were collected. IgG/IgM aPS/PT antibodies were assayed using commercial ELISA kit. Prevalences of IgG and IgM aPS/PT (pantiphospholipid antibody patients than in double and single positivity ones (p<0.0001 for all). APS/PT antibodies were associated to severe thrombosis, severe pregnancy complications inducing prematurity, and vascular microangiopathy, all generally associated to high risk APS forms requiring strong therapy.

  14. A three-layer immunoradiometric assay for determination of IgG subclass antibodies in Human Sera (''IgG subclass RAST'')

    International Nuclear Information System (INIS)

    Djurup, R.; Soendergaard, I.; Weeke, B.; University of Copenhagen, Denmark); Magnusson, C.G.M.

    1984-01-01

    We report the development of a three-layer immunoradiometric assay (TIRA) for measurement of IgG antibodies of all four subclasses in human sera. The first layer consists of diluted human serum, the second layer is monoclonal mouse antibodies to human IgG subclasses, and the third layer is 125 I-labelled rabbit anti-mouse IgG. Monoclonal anti-IgGI, anti-IgG3 and anti-IgG4 reacted only with their complementary IgG subclass, whereas the anti-IgG2 showed slight cross-reactivity to immunoglobins of other subclasses and classes and to light chain proteins. The observed cross-reactivity was found to be without importance, when the TIRA was applied to measurement of IgG subclass antibodies. Equipotency was established by use of appropriate dilutions of the monoclonal antibodies, and the assay was calibrated by use of human reference serum. The TIRA therefore permits reliable inter-individual and intra-individual comparisons of the IgG antibody response in all four subclasses. Non-specific binding obtained with pooled normal human serum was below 0.33%. Inter-assay coefficient of variation was between 18 and 27%. The TIRA was applied to measurement of IgG subclass antibodies to timothy grass pollen in sera from grass pollen allergies undergoing immunotherapy. (author)

  15. Comparison of Two Assays to Determine Anti-Citrullinated Peptide Antibodies in Rheumatoid Arthritis in relation to Other Chronic Inflammatory Rheumatic Diseases: Assaying Anti-Modified Citrullinated Vimentin Antibodies Adds Value to Second-Generation Anti-Citrullinated Cyclic Peptides Testing

    Science.gov (United States)

    Díaz-Toscano, Miriam Lizette; Olivas-Flores, Eva Maria; Zavaleta-Muñiz, Soraya Amali; Gamez-Nava, Jorge Ivan; Cardona-Muñoz, Ernesto German; Ponce-Guarneros, Manuel; Castro-Contreras, Uriel; Nava, Arnulfo; Salazar-Paramo, Mario; Celis, Alfredo; Fajardo-Robledo, Nicte Selene; Corona-Sanchez, Esther Guadalupe; Gonzalez-Lopez, Laura

    2014-01-01

    Determination of anti-citrullinated peptide antibodies (ACPA) plays a relevant role in the diagnosis of rheumatoid arthritis (RA). To date, it is still unclear if the use of several tests for these autoantibodies in the same patient offers additional value as compared to performing only one test. Therefore, we evaluated the performance of using two assays for ACPA: second-generation anti-citrullinated cyclic peptides antibodies (anti-CCP2) and anti-mutated citrullinated vimentin (anti-MCV) antibodies for the diagnosis of RA. We compared three groups: RA (n = 142), chronic inflammatory disease (CIRD, n = 86), and clinically healthy subjects (CHS, n = 56) to evaluate sensitivity, specificity, predictive values, and likelihood ratios (LR) of these two assays for the presence of RA. A lower frequency of positivity for anti-CCP2 was found in RA (66.2%) as compared with anti-MCV (81.0%). When comparing RA versus other CIRD, sensitivity increased when both assays were performed. This strategy of testing both assays had high specificity and LR+. We conclude that adding the assay of anti-MCV antibodies to the determination of anti-CCP2 increases the sensitivity for detecting seropositive RA. Therefore, we propose the use of both assays in the initial screening of RA in longitudinal studies, including early onset of undifferentiated arthritis. PMID:25025037

  16. Comparison of Two Assays to Determine Anti-Citrullinated Peptide Antibodies in Rheumatoid Arthritis in relation to Other Chronic Inflammatory Rheumatic Diseases: Assaying Anti-Modified Citrullinated Vimentin Antibodies Adds Value to Second-Generation Anti-Citrullinated Cyclic Peptides Testing

    Directory of Open Access Journals (Sweden)

    Miriam Lizette Díaz-Toscano

    2014-01-01

    Full Text Available Determination of anti-citrullinated peptide antibodies (ACPA plays a relevant role in the diagnosis of rheumatoid arthritis (RA. To date, it is still unclear if the use of several tests for these autoantibodies in the same patient offers additional value as compared to performing only one test. Therefore, we evaluated the performance of using two assays for ACPA: second-generation anti-citrullinated cyclic peptides antibodies (anti-CCP2 and anti-mutated citrullinated vimentin (anti-MCV antibodies for the diagnosis of RA. We compared three groups: RA (n=142, chronic inflammatory disease (CIRD, n=86, and clinically healthy subjects (CHS, n=56 to evaluate sensitivity, specificity, predictive values, and likelihood ratios (LR of these two assays for the presence of RA. A lower frequency of positivity for anti-CCP2 was found in RA (66.2% as compared with anti-MCV (81.0%. When comparing RA versus other CIRD, sensitivity increased when both assays were performed. This strategy of testing both assays had high specificity and LR+. We conclude that adding the assay of anti-MCV antibodies to the determination of anti-CCP2 increases the sensitivity for detecting seropositive RA. Therefore, we propose the use of both assays in the initial screening of RA in longitudinal studies, including early onset of undifferentiated arthritis.

  17. Detection of anti-Toxoplasma gondii antibodies in chronic myeloid leukemia and acute myeloid leukemia patients

    Directory of Open Access Journals (Sweden)

    Mohammad Javad Gharavi

    2017-09-01

    Full Text Available Background and Aim: Infection of Toxoplasma gondii is a worldwide distribution. Toxoplasmosis in patients who are immunocompromised by virtue of underlying leukemia disease has received relatively little attention. This study was aimed to evaluate IgG and IgM antibodies of T. gondii and to minimize the role of T. gondii and opportunistic infection complication at the early stage of infection in leukemia patients. Materials and Methods: The purpose of this assay was to measure anti-T. gondii IgG and IgM antibodies by enzyme-linked immunosorbent assay (ELISA technique in leukemia patients. Results: IgG antibodies against T. gondii were detected by ELISA in 96 (56.4% leukemia patients and 72 (42.4% control group. IgM antibodies were found in 10 patients (5.9% with leukemia and 3 (1.8% in the corresponding. Conclusion: Our finding indicated that leukemia patients under immunosuppressive condition should not be neglected. Toxoplasmosis in leukemia patients as a main risk factor is considered, meanwhile in some patients, due to possibility of the presence of secondary infection that leads to severe toxoplasmosis.

  18. Prevalence of Antibodies to Zika Virus in Mothers from Hawaii Who Delivered Babies with and without Microcephaly between 2009-2012.

    Directory of Open Access Journals (Sweden)

    Mukesh Kumar

    2016-12-01

    Full Text Available Zika virus (ZIKV is an emerging mosquito-borne pathogen. ZIKV infection is linked to the development of severe fetal abnormalities that include spontaneous abortion, stillbirth, hydranencephaly, and microcephaly. ZIKV outbreaks have been recorded in the United States. We recently demonstrated the first congenital ZIKV infection in the United States. In this study, we investigated archived blood samples from six mothers who gave birth to babies with microcephaly and 12 mothers who gave birth to healthy babies in Hawaii between 2009 and 2012. We tested maternal blood for the presence of ZIKV IgM and IgG antibodies using commercially available human ZIKV IgM and IgG ELISA kits. Blood from one mother who delivered babies with microcephaly tested positive for ZIKV IgM antibody (16.6% and blood from three mothers tested positive for ZIKV IgG antibody (50%. ZIKV showed a trend toward significance with microcephaly. ZIKV IgG antibody positive mothers were more likely to deliver babies with microcephaly than mothers who were negative for ZIKV IgG antibodies (Odds ratio [OR] = 11.0, 95% confidence interval [CI] = 0.8-147.9, p = 0.083. Similarly, ZIKV IgM antibody positive mothers were also more likely to deliver babies with microcephaly than mothers who were negative for ZIKV IgM antibody (OR = 6.8, 95% CI = 0.2-195.1. These data provide further evidence of a link between ZIKV infection and microcephaly and suggests presence of ZIKV positive cases and associated microcephaly in the United States as early as 2009.

  19. Le profil clinique et immunologique de 15 patients Marocains atteints de syndrome hyper IgM

    Science.gov (United States)

    Ouair, Hind; Benhsaien, Ibtihal; Jeddane, Leila; El Bakkouri, Jalila; Elhafidi, Naima; Rada, Noureddine; Najib, Jilali; Ailal, Fatima; Alj, Hanane Salih; Bousfiha, Ahmed Aziz

    2017-01-01

    Le Syndrome hyper IgM est un déficit immunitaire héréditaire bien connu, décrit pour la première fois en 1961. Il est causé par un défaut au niveau des lymphocytes B, caractérisé par un taux sérique normal ou élevé des IgM et un taux bas ou nul des IgG, IgA, IgE résultant d'une déficience de la commutation isotypique. Ses manifestations cliniques sont dominées par les infections à répétition, surtout du tube digestif, de la sphère ORL et des poumons. Le syndrome est causé par un défaut de commutation de classe d'immunoglobuline dans les cellules B, et une diminution de la capacité des monocytes à induire la prolifération des lymphocytes T. Le résultat net de tous ces défauts est la susceptibilité accrue aux infections opportunistes à Pneumocystis jiroveci, Cryptosporidium spp et d'autres organismes intracellulaires, ainsi qu'une fréquence élevée d'infections bactériennes et virales. L'intérêt de ce projet est d'illustrer l'importance de la compréhension des mécanismes physiopathologiques associés à cette susceptibilité accrue aux infections, ce qui permettra une meilleure prise en charge diagnostique et thérapeutique des patients atteint du Syndrome hyper IgM (SHIM). PMID:28690727

  20. Surveying the CGM and IGM across 4 orders of magnitude in environmental density

    Science.gov (United States)

    Burchett, Joseph

    2017-08-01

    Environment matters when it comes to galaxy evolution, and the mechanisms driving this evolution are reflected in the diffuse gas residing within the large-scale structures enveloping the cosmic galaxy population. QSO absorption lines effectively probe the circumgalactic medium (CGM) and intragroup and intracluster media, and work thus far hints at profound environmental effects on the CGM. However, sample sizes remain small, and a unifying picture of the gas characteristics across diverse environments has yet to emerge. Within the Sloan Digital Sky Survey, we have identified a sample volume containing a remarkable diversity in large-scale environment with an array of voids, >10,000 groups, several filaments, and 5 clusters, including the Coma Supercluster and CfA Great Wall. Leveraging the Hubble Spectroscopic Legacy Archive (HSLA), we propose a study using >360 background QSOs probing this volume to study the effects of large-scale environment on CGM and intergalactic medium (IGM) gas. The z = 0.019-0.028 spectroscopic galaxy sample is uniformly complete to galaxies L > 0.03 L* and, with the HSLA, produces 200 galaxy/sightline pairs within 300-kpc impact parameters across a wide range of environmental densities and structures.Upon quantifying the galaxy environment and identifying/measuring the QSO absorption lines at z = 0.019-0.028, we will pursue the following primary science goals:1. Constrain the CGM/IGM physical conditions across four orders of magnitude in galaxy density2. Compare ionic abundances and ionization states in the CGM of galaxies in filaments vs. voids3. Statistically investigate the IGM/CGM gas properties from structure to structure

  1. Soluble IgM links apoptosis to complement activation in early alcoholic liver disease in mice.

    Science.gov (United States)

    Smathers, Rebecca L; Chiang, Dian J; McMullen, Megan R; Feldstein, Ariel E; Roychowdhury, Sanjoy; Nagy, Laura E

    2016-04-01

    Ethanol feeding in mice activates complement via C1q binding to apoptotic cells in the liver; complement contributes to ethanol-induced inflammation and injury. Despite the critical role of C1q in ethanol-induced injury, the mechanism by which ethanol activates C1q remains poorly understood. Secretory IgM (sIgM), traditionally considered to act as an anti-microbial, also has critical housekeeping functions, facilitating clearance of apoptotic cells, at least in part through activation of C1q. Therefore, we hypothesized that (1) ethanol-induced apoptosis in the liver recruits sIgM, facilitating the activation of C1q and complement and (2) C1INH (C1 esterase inhibitor), which inhibits C1 functional activity, prevents complement activation and decreases ethanol-induced liver injury. Female C57BL/6 wild-type, C1qa(-/-), BID(-/-) and sIgM(-/-) mice were fed ethanol containing liquid diets or pair-fed control diets. C1INH or vehicle was given via tail vein injection to ethanol- or pair-fed wild-type mice at 24 and 48h prior to euthanasia. Ethanol exposure increased apoptosis in the liver, as well as the accumulation of IgM in the liver. In the early stages of ethanol feeding, C1q co-localized with IgM in the peri-sinusoidal space of the liver and accumulation of IgM and C3b was dependent on ethanol-induced BID-dependent apoptosis. sIgM(-/-) mice were protected from both ethanol-induced activation of complement and early ethanol-induced liver injury when compared to wild-type mice. Treatment with C1INH also decreased hepatic C3b deposition and ethanol-induced injury. These data indicate that sIgM contributes to activation of complement and ethanol-induced increases in inflammatory cytokine expression and hepatocyte injury in the early stages of ethanol-induced liver injury. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Retrospective analysis of dengue specific IgM reactive serum samples

    OpenAIRE

    Bhattacharya, Nemai; Bandyopadhyay, Bhaswati; Bhattacharjee, Indranil; Mukherjee, Hiranmoy; Talukdar, Srabani; Mondal, Ruby; Pramanick, Netai; Chandra, Goutam; Hati, Amiya K.

    2013-01-01

    Objective: To conduct a retrospective analysis of dengue cases in Kolkata, on the basis of presence of anti-dengue IgM in their sera and presence or absence of anti-dengue IgG and dengue specific Non structural 1 (NS1) antigen in each of the serum sample. Methods: Sample was tested quantitatively employing ELISA technique, using Biorad test kits, with a view to get a more comprehensive picture of dengue in an urban endemic area and also to evaluate individual cases. Results: Th...

  3. Study of a Cohort of 1,886 Persons To Determine Changes in Antibody Reactivity to Borrelia burgdorferi 3 Months after a Tick Bite

    DEFF Research Database (Denmark)

    Dessau, Ram B; Fryland, Linda; Wilhelmsson, Peter

    2015-01-01

    Lyme borreliosis is a tick-borne disease caused by the bacterium Borrelia burgdorferi. The most frequent clinical manifestation is a rash called erythema migrans. Changes in antibody reactivity to B. burgdorferi 3 months after a tick bite are measured using enzyme-linked immunosorbent assays...... and a 1.8-fold rise for the C6 assay. Of 1,886 persons, 102/101 (5.4%) had a significant rise in antibody reactivity in the flagellar assay or the C6 assay. Among 40 cases with a diagnosis of Lyme borreliosis, the sensitivities corresponding to a rise in antibodies were 33% and 50% for the flagellar...... in either ELISA but only 40 (2.1%) developed clinical Lyme borreliosis....

  4. Elevated serum antiphospholipid antibodies in adults with celiac disease.

    Science.gov (United States)

    Laine, Outi; Pitkänen, Katariina; Lindfors, Katri; Huhtala, Heini; Niemelä, Onni; Collin, Pekka; Kurppa, Kalle; Kaukinen, Katri

    2017-12-02

    An increased incidence of thrombosis is suggested in celiac disease. We explored serum levels of antiphospholipid antibodies in untreated and treated adult celiac disease patients. A cohort of 179 biopsy-proven celiac disease patients (89 untreated, 90 on long-term gluten-free diet) and 91 non-celiac controls underwent clinical examination, assessment of celiac serology and enzyme immunoassay testing for anticardiolipin IgG and IgM, prothrombin IgG, and phosphatidylserine-prothrombin IgG and IgM. The level of antiphospholipid antibodies was higher in celiac disease patients compared with controls: anticardiolipin IgG 4.9 (0.7-33.8) vs 2.2 (0.4-9.6) U/ml, antiprothrombin IgG 2.9 (0.3-87.8) vs 2.1 (0.5-187.0) U/ml, antiphosphatidylserine-prothrombin IgG 6.9 (0.0-54.1) vs 2.3 (0.5-15.1) U/ml; p celiac disease at presentation (gastrointestinal symptoms, malabsorption or anemia, and extraintestinal symptoms or screen-detected disease) had no effect on the level of serum antiphospholipid antibodies. The serum level of antiphospholipid antibodies is increased in adults with celiac disease. The higher level of antibodies in treated patients suggests that the increase is not gluten-dependent. The prothrombotic role of antiphospholipid antibodies in celiac disease warrants further studies. Copyright © 2017 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

  5. Highly sensitive and specific determination of mercury(II) ion in water, food and cosmetic samples with an ELISA based on a novel monoclonal antibody

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yuzhen; Li, Yuan [Sichuan University, College of Chemistry, Chengdu (China); Yang, Hong [Soochow University, College of Pharmacy, Suzhou (China); Pschenitza, Michael; Niessner, Reinhard; Knopp, Dietmar [Technical University Munich, Chair for Analytical Chemistry, Institute of Hydrochemistry and Chemical Balneology, Munich (Germany); Deng, Anping [Sichuan University, College of Chemistry, Chengdu (China); Soochow University, College of Chemistry, Chemical Engineering and Materials Science, Suzhou (China)

    2012-07-15

    Mercury is one of the most toxic heavy metals present in the environment. In this study, a highly sensitive and specific monoclonal antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (ELISA) for the determination of Hg{sup 2+} was developed. A new bifunctional ligand, 6-mercaptonicotinic acid (MNA), which contains a pyridine ring bearing a carboxylic group and a mercapto group, was selected for the preparation of immunogen. After immunization of mice and performing the hybridoma technique, the obtained mAb was characterized for its binding affinity and selectivity for Hg{sup 2+}. Based on this novel mAb, an ELISA was established. At optimal experimental conditions, the standard curve of the ELISA for Hg{sup 2+} was constructed in concentration range of 0.1-100 ng mL{sup -1}. The values of IC{sub 50} and LOD of the assay were found to be 1.12 and 0.08 ng mL{sup -1}. The cross-reactivity was lower than 2 % with MNA, CH{sub 3}Hg, and CH{sub 3}Hg-MNA and was 11.5 % and 4.4 % for Hg{sup +} and Au{sup 3+}, respectively. No cross-reactivity was found with other metal ions such as Cu{sup 2+}, Sn{sup 2+}, Ni{sup 2+}, Mn{sup 2+}, Pb{sup 2+}, Zn{sup 2+}, Cd{sup 2+}, Fe{sup 2+}, Co{sup 2+}, Mg{sup 2+}, Ca{sup 2+}, and anions such as Cl{sup -}, NO{sub 3} {sup -}, NO{sub 2} {sup -}, HCO{sub 3} {sup -}, F{sup -}, and SO{sub 4} {sup 2-}, indicating that the assay displays not only high sensitivity but also high selectivity. Different kinds of samples including water, milk, green vegetable, kelp, facial cleanser, and night cream were spiked with Hg{sup 2+} and the extracts were analyzed by ELISA. Acceptable recovery rates of 80.0-113.0 % and coefficients of variation of 1.9-18.6 % were obtained. A linear relationship between ELISA and cold-vapor atomic fluorescence spectroscopy (CV-AFS) as indicated by a correlation coefficient of 0.97 for liquid samples (water samples) and 0.98 for other samples was obtained. The proposed mAb-based ELISA provides a

  6. Determination of Anti-Adeno-Associated Viral Vector Neutralizing Antibodies in Patients With Heart Failure in the Cardiovascular Foundation of Colombia (ANVIAS): Study Protocol

    Science.gov (United States)

    Prada, Carlos E; Lopez, Marcos; Castillo, Victor; Echeverria, Luis Eduardo; Serrano, Norma

    2016-01-01

    Background Recent progress in the pathophysiology of heart failure (HF) has led to the development of new therapeutic options such as gene therapy and the use of adeno-associated viral (AAV) vectors. Despite the promising results in early clinical trials of gene therapy for HF, various obstacles have been faced, such as the presence of neutralizing antibodies (NAbs) against the capsid vectors. NAb activity limits vector transduction levels and therefore diminishes the final therapeutic response. Recent studies evaluating the prevalence of NAbs in various populations found considerable geographic variability for each AAV serotype. However, the levels of NAbs in Latin American populations are unknown, becoming a limiting factor to conducting AAV vector therapeutic trials in this population. Objective The goal of this study is to determine for the first time, the prevalence of anti-AAV NAbs for the serotypes 1, 2, and 9 in HF patients from the city of Bucaramanga, Colombia, using the in vitro transduction inhibition assay. Methods We will conduct a cross-sectional study with patients who periodically attend the HF clinic of the Cardiovascular Foundation of Colombia and healthy volunteers matched for age and sex. For all participants, we will evaluate the NAb levels against serotypes AAV1, AAV2, and AAV9. We will determine NAb levels using the in vitro transduction inhibition assay. In addition, participants will answer a survey to evaluate their epidemiological and socioeconomic variables. Participation in the study will be voluntary and all participants will sign an informed consent document before any intervention. Results The project is in the first phase: elaboration of case report forms and the informed consent form, and design of the recruitment strategy. Patient recruitment is expected to begin in the spring of 2016. We expect to have preliminary results, including the titer of the viral vectors, multiplicity of infections that we will use for each serotype

  7. Characterization of the mechanism of protection mediated by CS-D7, a monoclonal antibody to Staphylococcus aureus iron regulated surface determinant B (IsdB

    Directory of Open Access Journals (Sweden)

    Gregory ePancari

    2012-03-01

    Full Text Available We previously reported the development of a human monoclonal antibody (CS-D7, IgG1 with specificity and affinity for the iron regulated surface determinant B (IsdB of Staphylococcus aureus. CS-D7 mediates opsonophagocytic killing in vitro and protection in a murine sepsis model. In light of recent data indicating that IsdB specific T cells (CD4+, Th17, not Ab, mediate protection after vaccination with IsdB, it is important to investigate the mechanism of protection mediated by CS-D7. The mAb was examined to determine if it blocked heme binding to IsdB in vitro. The mAb was not found to have heme blocking activity, nor did it prevent bacterial growth under in vivo conditions, in an implanted growth chamber. To assess the role of the mAb Fc a point mutation was introduced at aa 297 (CS-D7●N297A. This point mutation removes Fc effector functions. In vitro analysis of the mutein confirmed that it lacked measurable binding to FcγR, and that it did not fix complement. The mutein had dramatically reduced in vitro opsonic OP activity compared to CS-D7. Nonetheless, the mutein conferred protection equivalent to the wild type mAb in the murine sepsis model. Both wild type and mutein mAbs were efficacious in FcγR deletion mice (including both FcγRII-/- mice and FcγRIII-/- mice, indicating that these receptors were not essential for mAb mediated protection in vivo. Protection mediated by CS-D7 was lost in Balb/c mice depleted of C3 with cobra venom factor (CFV, was lost in mice depleted of superoxide dismutase (SOD in P47phox deletion mice, and was absent in SCID mice. Enhanced clearance of S. aureus in the liver of CS-D7 treated mice and enhanced production of INF-γ, but not of IL17, may play a role in the mechanism of protection mediated by the mAb. CS-D7 apparently mediates survival in challenged mice through a mechanism involving complement, phagocytes, and lymphocytes, but which does not depend on interaction with FcγR, or on blocking heme

  8. Antiganglioside antibodies in Guillain-Barré syndrome after a recent cytomegalovirus infection.

    Science.gov (United States)

    Khalili-Shirazi, A; Gregson, N; Gray, I; Rees, J; Winer, J; Hughes, R

    1999-03-01

    To study the association between anti-ganglioside antibody responses and Guillan-Barré syndrome (GBS) after a recent cytomegalovirus (CMV) infection. Enzyme linked immunosorbant assay (ELISA) was undertaken on serum samples from 14 patients with GBS with recent cytomegalovirus (CMV) infection (CMV+GBS) and 12 without (CMV-GBS), 17 patients with other neurological diseases (OND), 11 patients with a recent CMV infection but without neurological involvement, 11 patients with recent Epstein-Barr virus (EBV) infection but without neurological involvement, and 20 normal control (NC) subjects. IgM antibodies were found at 1:100 serum dilution to gangliosides GM2 (six of 14 patients), GM1 (four of 14), GD1a (three of 14) and GD1b (two of 14) in the serum samples of the CMV+GBS patients, but not in those of any of the CMV-GBS patients. IgM antibodies were also found to gangliosides GM1, GD1a, and GD1b in one of 11 OND patients, to ganglioside GM1 in one of 11 non- neurological CMV patients, and to ganglioside GD1b in one of 20 NC subjects. Some patients with EBV infection had IgM antibodies to gangliosides GM1 (five of 11), GM2 (three of 11), and GD1a (two of 11). However, the antibodies to ganglioside GM2 had a low titre, none being positive at 1:200 dilution, whereas five of the CMV+GBS serum samples remained positive at this dilution. Antibodies to ganglioside GM2 are often associated with GBS after CMV infection, but their relevance is not known. It is unlikely that CMV infection and anti-ganglioside GM2 antibodies are solely responsible and an additional factor is required to elicit GBS.

  9. Synthetic peptides for efficient discrimination of anti-enterovirus antibodies at the serotype level.

    Science.gov (United States)

    Routsias, John G; Mavrouli, Maria D; Antonaki, Georgia; Spanakis, Nikolaos; Tsakris, Athanassios

    2014-08-01

    Enteroviruses are important human pathogens, causing a broad spectrum of diseases from minor common colds to fatal myocarditis. However, certain disease syndromes are caused by one or few serotypes. Serotype identification is difficult due to the laborious neutralization tests that lack of sensitivity, while in commercial ELISAs homotypic antibodies' activities are largely masked by the recognition of genera-specific epitopes by heterotypic antibodies. In the present study homotypic assays were developed with the ability to discriminate different enterovirus serotypes. Seventy-three children sera, positive for IgM antibodies against enterovirus genus and 49 healthy children were examined for the presence of antibodies against 14 synthetic peptides derived from a non-conserved region of the VP1 protein of coxsackieviruses B2, B3, B4, B5, A9, A16, A24, echoviruses 6, 7, 9, 11, 30, enterovirus 71 and parechovirus 1. 50% of the anti-enterovirus IgM positive sera (>150 BU) reacted with the peptides with the majority of them to preferentially recognize one of them, supporting the homotypic nature of our assay. Inhibition studies yielded homologous inhibition rates 67-95% suggesting that specific peptide recognition actually occurred. The diagnostic value of our assay was tested in blood samples drawn over a 1.5-year period from a 5-year old patient. The anti-enterovirus reactivity was clearly attributed to echovirus serotype 11. The IgM/IgG antibody ratio was reversed 4 months later and subsequently IgM antibodies dropped below the cutoff point. In this paper we demonstrate that our assay can be used to discriminate between antibodies targeting different enterovirus serotypes. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Characterization of antibodies against ferret immunoglobulins, cytokines and CD markers

    DEFF Research Database (Denmark)

    Martel, Cyril Jean-Marie; Aasted, Bent

    2009-01-01

    Ferret IgG and IgM were purified from normal serum, while ferret IgA was purified from bile. The estimated molecular weights of the immunoglobulin gamma, alpha and mu heavy chains were found to be 54 kDa, 69 kDa and 83 kDa, respectively. For immunological (ELISA) quantification of ferret...... immunoglobulins, we identified and characterized polyclonal antibodies towards ferret IgG, IgM and IgA. We also identified 22 monoclonal antibodies (mAbs) raised mostly against human CD markers which cross-reacted with ferret leukocytes. These antibodies were originally specific against human CD8, CD9, CD14, CD18......, CD25, CD29, CD32, CD44, CD61, CD71, CD79b, CD88, CD104, CD172a and mink CD3. Finally, we identified 4 cross-reacting mAbs with specificities against ferret interferon-gamma, TNF-alpha, interleukin-4 and interleukin-8....

  11. Cold-induced precipitation of a monoclonal IgM: a negative activation enthalpy reaction.

    Science.gov (United States)

    Meliga, Stefano C; Farrugia, William; Ramsland, Paul A; Falconer, Robert J

    2013-01-17

    Cold-induced precipitation of a monoclonal IgM cryoglobulin isolated from a patient with Waldenström's macroglobulinemia was observed to have a negative activation enthalpy. The rate of the reaction increased, as the temperature decreased. Differential scanning calorimetry of the monoclonal IgM showed precipitation as an inverted peak during a downward temperature scan. The transition temperature was between 14 and 15 °C and was possibly concentration dependent. At temperatures below the transition the precipitation was best described by second-order kinetics. The difference in change in enthalpy between precipitation and disassociation suggests that cold-induced precipitation had a fast precipitation stage followed by a slower consolidation reaction. Negligible curvature of the Eyring plot suggested the precipitation reaction was dominated by van der Waal forces and hydrogen bonding. Conversely, during an upward temperature scan, disassociation was observed as a positive enthalpy peak. This reaction had two stages, a reaction undoing consolidation followed by heat-induced disassociation that had first-order kinetics.

  12. Retrospective analysis of dengue specific IgM reactive serum samples

    Directory of Open Access Journals (Sweden)

    Nemai Bhattacharya

    2013-04-01

    Full Text Available Objective: To conduct a retrospective analysis of dengue cases in Kolkata, on the basis of presence of anti-dengue IgM in their sera and presence or absence of anti-dengue IgG and dengue specific Non structural 1 (NS1 a