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Sample records for phytohemagglutinin

  1. The Significance Application of Indigenous Phytohemagglutinin (PHA) Mitogen on Metaphase and Cell Culture Procedure.

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    Movafagh, Abolfazl; Heydary, Hassan; Mortazavi-Tabatabaei, Seyed Abdolreza; Azargashb, Eznollah

    2011-01-01

    Phytohemagglutinin (PHA) is a lectin, obtained from the red kidney bean that binds to the membranes of T-cells and stimulates metabolic activity, cell division, etc. The object of this research was the comparison between self made PHA (Indigenous) and imported commercial one, following conventional and High Resolution Cell Synchronization technique (HRCS) .From each blood sample of healthy individual donor replicate cell culture with two different PHA (self-made and commercial imported) with same concentration were cultured simultaneously. For culture cells, 3-5 × 106(6) cells were cultured in 4 mL medium( RPMI 1640 supplemented with 15 per cent heat inactivated fetal bovine serum, 0.1 mL Phytohemagglutinin was added and kept at 37°C in an atmosphere containing 5% CO2. The processing of mitotic division from 48 h and 72 h cultures was performed according to the standard and High Resolution Cell Synchronization technique. Cytogenetic studies were performed in 100 normal healthy blood donor individuals. Statistical analysis was performed by SPSS (version 16, Inc.USA) software.Our results indicate that the preparation of fresh Phytohemagglutinin at the time of cell division and cell culture procedure reveals satisfactory score. The overall frequency of mitotic index in our study was better when compared with commercial imported Phytohemagglutinin (p < 0.001).The significant differences in the results may be due to fresh preparation. However, cost effective, easy and nearest approach of this indigenous product and high demand for this product among health care services can be considered.

  2. [Elimination of large pyroninophilic cells due to the effect of phytohemagglutinin].

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    Bykovskaia, S N; Bykovskiĭ, A F; Shepelenko, A M

    1975-09-01

    Large pyroninophilic lymphocytes adsorbed on the surface of target-cells disappeared after phytohemagglutinin (PHA) addition. Incubation during 45 minutes in the presence of PHA did not reveal cisternas of the granular endoplasmatic reticulum and the number of mitochondrias decreased. The H3-thymidine-labeled cells were almost eliminated. There appeared population of small lymphocytes with even outline and clear cytoplasm, poor in organellas, with ribosomas freely scattered in it. After 24--48 hours of incubation they transformed into blasts, large cells with clear nucleus and clear cytoplasm in which no cisternas of granular endoplasmatic reticulum were revealed.

  3. [Gamma interferon induced in human leukocytes by phytohemagglutinin: its production and biological characteristics].

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    Danielescu, G; Maniu, H; Georgescu, T; Cajal, N

    1988-01-01

    Human gamma type interferon (IFN) preparations were obtained through phytohemagglutinin stimulation of leukocytes from the peripheral blood. Biological value of these preparations varied between 160 u and 800 u/ml, depending on leukocyte incubation medium, culture system and inductor conservation. The rising of the antiviral activity through association between gamma (3 u) and alpha (27 u) interferons was revealed by the virus quantity reduction (in this case the vesicular stomatitis virus was used) during a 24-hour multiplication cycle. The protection ensured by the mixture of the two types of interferon was about ten times higher than the additive effect of the two preparations. Study of the antiproliferative activity of a gamma interferon preparation was conducted on two human cell lines of tumoral origin (T-10 from a glioblastoma, and HEp-2) and revealed the difficulties to quantify precisely this property of the crude gamma interferon preparations.

  4. Evaluation by hierarchical clustering of multiple cytokine expression after phytohemagglutinin stimulation

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    Yang Chunhe

    2016-01-01

    Full Text Available The hierarchical clustering method has been used for exploration of gene expression and proteomic profiles; however, little research into its application in the examination of expression of multiplecytokine/chemokine responses to stimuli has been reported. Thus, little progress has been made on how phytohemagglutinin(PHA affects cytokine expression profiling on a large scale in the human hematological system. To investigate the characteristic expression pattern under PHA stimulation, Luminex, a multiplex bead-based suspension array, was performed. The data set collected from human peripheral blood mononuclear cells (PBMC was analyzed using the hierarchical clustering method. It was revealed that two specific chemokines (CCL3 andCCL4 underwent significantly greater quantitative changes during induction of expression than other tested cytokines/chemokines after PHA stimulation. This result indicates that hierarchical clustering is a useful tool for detecting fine patterns during exploration of biological data, and that it can play an important role in comparative studies.

  5. Phytohemagglutinins augment red kidney bean (Phaseolus vulgaris L.) induced allergic manifestations.

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    Kumar, Sandeep; Verma, Alok Kumar; Sharma, Akanksha; Kumar, Dinesh; Tripathi, Anurag; Chaudhari, B P; Das, Mukul; Jain, S K; Dwivedi, Premendra D

    2013-11-20

    Red kidney bean (Phaseolus vulgaris L.), a commonly consumed bean has been reported to induce allergic reactions in susceptible individuals. Phytohemagglutinins (PHAs, mainly PHA-P) contribute a major proportion of red kidney bean seeds. However, their roles in red kidney bean induced allergic reactions are still to be explored. This study was carried out to understand the role of PHAs in allergic manifestations using BALB/c mice and cultures of splenocyte, RBL-2H3 cells as well as bone marrow mast cells (BMMCs). Also, the characterization of allergic components from PHA-P was studied by LC-MS/MS. Enhanced levels of specific IgE and IgG1, clinical scores, cytokines and chemokines, β-hexosaminidase, histamine, cysteinyl leukotriene, prostaglandin D2 and abrupt histological changes in the intestine, lung and spleen indicated a pivotal role of PHA-P in red kidney bean allergy. Further, LC-MS/MS study revealed two IgE binding components of PHA-P as PHA-L and PHA-E. Enhanced specific IgE/IgG1 and β-hexosaminidase level elucidated the possible role of PHA-L and PHA-E in allergic manifestations. Furthermore, in the presence of IgE inhibitor piceatannol, reduced β-hexosaminidase release to some extent was noticed. The up regulated expression of GATA-3 and T-bet expression was observed in PHA-L as well as PHA-E groups. Taken together, this study revealed the fact that allergenicity potential of red kidney bean may get augmented due to the presence of different phytohemagglutinins. Although food allergy is an immune provocation induced mainly by dietary allergenic protein components of the food, the role of dietary lectins in the food induced allergic manifestations cannot be ruled out. Here we provide the systematic evidences about the allergenic potential of PHAs and further disclosed the culprit components as PHA-L and PHA-E. It is an important finding that the PHA-L and PHA-E can cause allergic manifestations via not only the IgE mediated pathway but also the non

  6. Philadelphia chromosome detection in chronic myeloid leukemia: Utility of phytohemagglutinin-stimulated peripheral blood culture

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    Man Updesh Singh Sachdeva

    2012-01-01

    Full Text Available Background: The conventional cytogenetic approach to demonstrate Philadelphia (Ph chromosome at times does not yield enough number of metaphases or are of suboptimal quality. Further, the rapid molecular tests have completely pushed this simple technique into disrepute. Aims: This study aimed to evaluate usefulness of phytohemagglutinin (PHA-stimulated peripheral blood culture for detection of Ph chromosome in chronic myeloid leukemia (CML patients. Materials and Methods: Fifty-six patients, including 11 newly diagnosed cases of CML and 45 patients of CML on imatinib therapy showing the presence of Ph chromosome in unstimulated samples, were included in the study. Cytogenetic analysis was done on unstimulated samples, i.e. bone marrow aspirate, 24- and 48-h peripheral blood culture, and compared with PHA-stimulated 72-h peripheral blood culture. Results: The preparations from PHA-stimulated peripheral blood culture samples in all 56 patients yielded high number of good-quality metaphases. All the 11 (100% newly diagnosed patients and 39/45 (87% of the patients on imatinib therapy showed the presence of Ph chromosome in PHA-stimulated samples. Addition of PHA-stimulated 72-h peripheral blood culture preparation can be of use for increasing the diagnostic yield in cases of CML with suboptimal results on conventional cytogenetics from bone marrow aspirate sample.

  7. Isolectins of phytohemagglutinin are able to induce apoptosis in HEp-2 carcinoma cells in vitro.

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    Kochubei, T O; Maksymchuk, O V; Piven, O O; Lukash, L L

    2015-06-01

    To study the effects of total phytohemagglutinin (PHA) and its isolectins on cell death and apoptosis in human HEp-2 carcinoma cells and to analyze the possible molecular mechanisms of lectin induced apoptosis. The commercial preparation of the kidney beans (Phaseolus vulgaris) lectins and HEp-2 cells were used. Apoptosis index was determined using acridine orange and ethidium bromide staining. The expression levels of apoptosis mediator cleaved caspase-3 and proapoptotic Bax protein were studied by Western blot analysis. The gene expression levels were analyzed by qPCR. PHA and its isolectins induced apoptosis in HEp-2 cells accompanied by the increased expression of caspase-3 cleaved form, with PHA-E being the most effective. The treatment of HEp-2 cells with PHA or its isolectins resulted in a marked increase of Bax on both mRNA and protein levels. PHA and its isolectins were shown to induce the apoptosis in human HEp-2 carcinoma cells via increasing proapoptotic protein Bax and activating caspases-3.

  8. Genetic selection of mice for quantitative responsiveness of lymphocytes to phytohemagglutinin.

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    Stiffel, C; Liacopoulos-Briot, M; Decreusefond, C; Lambert, F

    1977-05-01

    A two-way selection was performed in mice according to the quantitative in vitro response of lymph node lymphocytes to the mitogenic activity of phytohemagglutinin (PHA). The foundation population was composed of outbred mice produced by reciprocal mating of equal numbers of mice from four different colonies. The selective breeding was carried out by mating of mice at each generation giving the best or the lowest response, respectively. The progressive interline separation produced by 6 generations of selective breeding demonstrates that responsiveness to PHA is submitted to polygenic regulation. The heritability of the character investigated is 0.28 +/- 0.08. The interline separation is also found with another T mitogen, concanavalin A (Con A). In spleen cells PHA and Con A produce a similar interline difference. In contrast, the purified protein derivative of tuberculin (PPD) stimulated both lines equally, and E. coli lipopolysaccharide gave only a slightly higher response in high line. This finding implies that our selection based upon response to PHA did not influence B cell function.

  9. Neutrophil-to-Lymphocyte Ratio Is Associated with Impaired Interferon-Gamma Release to Phytohemagglutinin.

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    Kwang-Sook Woo

    Full Text Available The neutrophil-to-lymphocyte ratio (NLR has been shown to predict adverse outcomes in several pathologic conditions. The majority of indeterminate interferon (IFN-γ release assays were due to inadequate IFN-γ response to the phytohemagglutinin. We sought to study the value of NLR to predict an indeterminate result of QuantiFERON-TB Gold In-Tube (QFT-GIT performed in routine laboratory practice.Results from 2,773 QFT-GIT assays were analyzed. Data collection included demographic data, the level of IFN-γ to nil, mitogen, and TB antigen of QFT-GIT, total WBC, and a differential count. We calculated the absolute neutrophil count, lymphocyte count, and NLR.Of the total, 224 (8.1% indeterminate results were observed. Twelve (1.8% showed indeterminate results in the NLR range from 1.71 to 2.84, but 132 (19.2% had indeterminate results in NLR ≥ 5.18 (p < 0.0001. The likelihood ratio for indeterminate results were 2.70 (95% CI, 2.36-3.08 in NLR ≥ 5.18 and 1.93 (95% CI, 1.64-2.27 in lymphocyte count ≤ 1050/μL. NLR and neutrophil count were independent predictors for indeterminate QFT-GIT result in multiple regression analysis. The IFN-γ response to PHA was negatively associated with NLR (r = -0.33, p < 0.001.We showed that the NLR is an independent predictor of indeterminate QFT-GIT result. Low frequency of indeterminate results in group with normal NLR may imply the importance of a balance between two cellular compartments in physiological and pathological conditions.

  10. Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

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    Cunha, E.V.; Costa, J.J.N.; Rossi, R.O.D.S.; Silva, A.W.B.; Passos, J.R.S.; Portela, A.M.L.R.; Pereira, D.C.S.T. [Núcleo de Biotecnologia de Sobral, NUBIS, Universidade Federal do Ceará, Sobral, CE (Brazil); Donato, M.A.M. [Laboratório de Ultraestrutura, CPqAM/FIOCRUZ, Universidade Federal de Pernambuco, Recife, PE (Brazil); Campello, C.C. [Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais, Faculdade de Medicina Veterinária, Universidade Estadual do Ceará, Fortaleza, CE (Brazil); Saraiva, M.V.A. [Núcleo de Biotecnologia de Sobral, NUBIS, Universidade Federal do Ceará, Sobral, CE (Brazil); Peixoto, C.A. [Laboratório de Ultraestrutura, CPqAM/FIOCRUZ, Universidade Federal de Pernambuco, Recife, PE (Brazil); Silva, J.R.V.; Santos, R.P. [Núcleo de Biotecnologia de Sobral, NUBIS, Universidade Federal do Ceará, Sobral, CE (Brazil)

    2013-03-19

    The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM{sup +} supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM{sup +} (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM{sup +} (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

  11. Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus preantral follicles

    Directory of Open Access Journals (Sweden)

    E.V. Cunha

    Full Text Available The objective this study was to determine the effect of phytohemagglutinin (PHA on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL. After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R, proliferating cell nuclear antigen (PCNA, and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%; 1 µg/mL PHA (96.43%; 10 µg/mL PHA (84.85%; 50 µg/mL PHA (85.29%; 100 µg/mL PHA (88.57%, and 200 µg/mL PHA (87.50], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21% when compared with control (5.41%, P < 0.05 and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1 and PCNA (4.4 ± 0.2 was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1. In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

  12. Immune responsiveness to phytohemagglutinin displays species but not sex differences in three anuran species

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    Jin, Chenchen; Qu, Kangshan; Caviedes-Vidal, Enrique

    2017-01-01

    Phytohemagglutinin (PHA)-induced skin swelling response is widely used as a rough surrogate of integrative cell-mediated and innate immunity across multiple vertebrate taxa due to its simplification and feasibility. However, little is known whether there are sex and interspecific differences of immune responsiveness to PHA in ectotherms, especially for anurans. Therefore, we studied sex and species differences of PHA response in three anurans, Asiatic toads (Bufo gargarizans), Dark-spotted frogs (Pelophylax nigromaculatus) and Mongolian toads (Pseudepidalea raddei), captured in northern regions of Anhui Province (China). Footpad thickness was measured prior to (0 h) and after (6, 12, 24, 48 and 72 h) a PHA injection and normalized against saline injection in the opposite footpad. Body mass was recorded at the beginning (0 h) and end of each assay (72 h). Results showed effects of PHA assay, sex and taxa on body mass. Relative maximum swelling response (PHAmax) ranged from 18.58–29.75%, 9.77 to 20.56% and 21.97 to 31.78% and its occurrence over time was apparent 10.6–19.72 h , 7.74–14.01 h and 17.39–23.94 h postinjection for Asiatic toads, Dark-spotted frogs and Mongolian toads, respectively. Finally, the magnitude or timing of PHAmax in Dark-spotted frogs was significantly thinner and faster than in Mongolian toads, and Asiatic toads had an in-between value, not different from the other two species. The magnitude of PHAmax was significantly positively correlated with the timing of PHAmax considering individuals altogether, but not when analyzed within species. Our results indicate that male and female anuran species respond similarly to PHA antigen stimulation, but the magnitude and timing of PHAmax is species-specific. Briefly, we provide new evidence for the suitability of PHA assay in non-model anuran species with different body sizes, and exhort the need to further investigate the nature of PHA assay at the hematological and histological levels in order

  13. Rapid diagnosis of vitamin D-dependent rickets type II by use of phytohemagglutinin-stimulated lymphocytes.

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    Takeda, E; Kuroda, Y; Saijo, T; Toshima, K; Naito, E; Kobashi, H; Iwakuni, Y; Miyao, M

    1986-03-28

    The interactions of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] with phytohemagglutinin (PHA)-stimulated lymphocytes from normal subjects and three patients with vitamin D-dependent rickets (DDR) type II were investigated. Impaired nuclear uptake and normal cytosol binding of [3H]1,25-(OH)2D3 were observed with PHA-stimulated lymphocytes of these patients as with their cultured skin fibroblasts. Furthermore, the incorporation of [14C]thymidine into PHA-stimulated lymphocytes of the patients was not reduced by 1,25-(OH)2D3, which is known to inhibit proliferation of various cells. These findings suggest that 1,25-(OH)2D3 receptors are reduced or absent in patients with DDR type II. Thus, the capacities of cytosol binding and nuclear uptake of 1,25-(OH)2D3 in PHA-stimulated lymphocytes seem to reflect those of endo-organs such as the intestine and bone. These findings show that a test of the effect of 1,25-(OH)2D3 on thymidine incorporation into PHA-stimulated lymphocytes is useful for rapid diagnosis of DDR type II.

  14. Identification and Characterization of Phytohemagglutinins from White Kidney Beans (Phaseolus vulgaris L., var. Beldia) in the Rat Small Intestine.

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    Nciri, Nader; Cho, Namjun; El Mhamdi, Faiçal; Ben Mansour, Abderraouf; Haj Sassi, Fayçal; Ben Aissa-Fennira, Fatma

    2016-01-01

    Although kidney bean (Phaseolus vulgaris L.) lectin toxicity is widely known, its effects in the gastrointestinal tract require further study. This investigation aimed to identify and characterize phytohemagglutinins (PHAs) in the small intestine and sera of rats following oral challenge with ground white beans. Twenty young, adult male rats were divided randomly into two groups of 10 animals each. The control group underwent gavage with a suspension of 300 mg of rodent pellet flour. The experimental group was administered a 300 mg Beldia bean flour suspension (BBFS). After 10 days of daily treatment, jejunal rinse liquid (JRL) and ileum rinse liquid and secretions, as well as sera, were collected. All biological fluids were screened for lectin reactivity using competitive inhibition ELISA, Ouchterlony double immunodiffusion, and immunoelectrophoresis techniques. The results revealed the presence of immunogenic intraluminal PHAs 3-4 h after the oral intake of the BBFS in the JRLs as well as in the jejunal and ileal secretions; however, no PHA was detectable in the rat sera. Ingestion of raw Beldia beans may lead to interaction between PHAs and the mucosa of the small intestine, potentially resulting in an inflammatory response.

  15. Delayed-type hypersensitivity, contact sensitivity, and phytohemagglutinin skin-test responses of heat- and cold-stressed calves.

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    Kelley, K W; Greenfield, R E; Evermann, J F; Parish, S M; Perryman, L E

    1982-05-01

    Three-week-old Holstein bull calves were used to investigate the effect of a 2-week chronic heat (35 C) or cold (-5 C) exposure on delayed-type hypersensitivity (DTH) reactions to purified protein derivative after sensitization with heat-killed Mycobacterium tuberculosis, contact sensitivity (CS) reactions to 1-fluoro-2,4-dinitrobenzene, and phytohemagglutinin (PHA) skin tests. Heat exposure reduced expression of DTH reactions by 42% and CS reactions by 38% at 24 hours after elicitation of the responses. The PHA-induced skin tests were not affected after 1 week of heat exposure, but this reaction was reduced by 20% after 2 weeks of heat exposure. The immune response of calves exposed to cold air temperatures was more complex. Cold exposure suppressed CS reactions by 39% at the end of both the 1st and 2nd weeks. The PHA response was reduced by 39% after 2 weeks of cold exposure. The DTH response depended on duration of cold exposure. The DTH reaction was increased by 42% after 1 week, but was reduced by 14% after 2 weeks. These data are consistent with the hypothesis that environmental stressors alter host resistance by affecting the immune system. Furthermore, these stress-induced changes in immune events depend on the type of immune response, the nature of the environmental stressor, and the length of time that calves are exposed to the stressor.

  16. Mifepristone acts as progesterone antagonist of non-genomic responses but inhibits phytohemagglutinin-induced proliferation in human T cells.

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    Chien, C H; Lai, J N; Liao, C F; Wang, O Y; Lu, L M; Huang, M I; Lee, W F; Shie, M C; Chien, E J

    2009-08-01

    Progesterone is an endogenous immunomodulator that suppresses T cell activation during pregnancy. The stimulation of membrane progesterone receptors (mPRs) would seem to be the cause of rapid non-genomic responses in human peripheral T cells, such as an elevation of intracellular calcium ([Ca(2+)](i)) and decreased intracellular pH (pH(i)). Mifepristone (RU486) produces mixed agonist/antagonist effects on immune cells compared with progesterone. We explored whether RU486 is an antagonist to mPRs and can block rapid non-genomic responses and the induction by phytohemagglutinin (PHA) of cell proliferation. Human male peripheral T cell responses in terms of pH(i) and [Ca(2+)](i) changes were measured using the fluorescent dyes, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) and fura-2, respectively. Expression of mPR mRNA was determined by RT-PCR analysis. Cell proliferation and cell toxicity were determined by [(3)H]-thymidine incorporation and MTT assay, respectively. The mRNAs of mPRalpha, mPRbeta and mPRgamma were expressed in T cells. RU486 blocked progesterone-mediated rapid responses including, the [Ca(2+)](i) increase and pH(i) decrease, in a dose related manner. RU486 did not block, but enhanced, the inhibitory effect of progesterone on PHA induced cell proliferation. RU486 alone inhibited proliferation induced by PHA and at >25 microM seems to be cytotoxic against resting T cells (P < 0.01). RU486 is antagonistic to the rapid mPR-mediated non-genomic responses, but is synergistic with progesterone with respect to the inhibition of PHA-induced cell proliferation. Our findings shine new light on RU486's clinical application and how this relates to the non-genomic rapid physiological responses caused by progesterone.

  17. Three-Color Flow Cytometry Detection of Intracellular Cytokines in Peripheral Blood Mononuclear Cells: Comparative Analysis of Phorbol Myristate Acetate-Ionomycin and Phytohemagglutinin Stimulation

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    Baran, Jarołsaw; Kowalczyk, Danuta; Ożóg, Mariola; Zembala, Marek

    2001-01-01

    The assessment of intracellular cytokines at the single-cell level by flow cytometry has recently become a potent tool in many areas of cell biology and in defining the role of cytokines in various human diseases. Three-color flow cytometry for detection of intracellular cytokines combined with simultaneous determination of lymphocytes (CD3+ and CD4+) or monocytes (CD33+ and CD14+) was used for comparison of phytohemagglutinin (PHA)-and phorbol myristate acetate (PMA)-ionomycin-induced production of intracellular cytokines in peripheral blood mononuclear cells (PBMCs) of healthy donors. We found that the number of PBMCs stained for tumor necrosis factor alpha and gamma interferon after 6 h of activation was higher when PMA-ionomycin was used for stimulation, while the frequencies of cells positive for interleukin 4 (IL-4) were similar for both stimulators. However, PMA-ionomycin stimulation caused prominent alterations of cell morphology and membrane expression of CD4 and CD14. In contrast, PHA did not cause downregulation of surface markers and resulted in less pronounced alterations in both forward and side scatter signals during flow cytometry analysis. Moreover, during 48 h of culture PHA stimulated tumor necrosis factor beta and IL-10 production, which was not observed when PMA-ionomycin was used. We conclude that the use of PHA for cell activation may limit in vitro artifacts and allow more precise analysis of intracellular cytokine production in various disease states. PMID:11238213

  18. The rapid immunosuppression in phytohemagglutinin-activated human T cells is inhibited by the proliferative Ca(2+) influx induced by progesterone and analogs.

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    Lin, Veronica Hui-Chen; Chen, Jiann-Jong; Liao, Chen-Chung; Lee, Shinn-Shing; Chien, Eileen Jea

    2016-07-01

    Progesterone, an endogenous immunomodulator, suppresses human T-cell activation during pregnancy. A sustained Ca(2 +) influx is an important signal for T-cell proliferation after crosslinking of T-cell receptor/CD3 complexes by anti-CD3 antibodies or phytohemagglutinin (PHA). Progesterone targets cell membrane sites inducing rapid responses including elevated intracellular free calcium concentration ([Ca(2+)]i) and suppressed T-cell PHA-activated proliferation. Interestingly, both PHA and progesterone induce [Ca(2+)]i elevation, but it remains unclear whether the PHA-induced Ca(2+) influx is affected by progesterone leading to T-cell immunosuppression. Primary T-cells were isolated from human peripheral blood and the quench effect on intracellular fura-2 fluorescence of Mn(2+) was used to explore the responses to Ca(2+) influx with cell proliferation being determined by MTT assay. PHA-stimulated Ca(2+) influx was dose-dependently suppressed by progesterone and its agonist R5020, which correlated with PHA-activated T-cell proliferation inhibition. A similar dose-dependent suppression effect on cellular Ca(2+) influx and proliferation occurred with the TRPC channel inhibitor BTP2 and selective TRPC3 channel inhibitor Pyr3. In addition, two progesterone analogs, Org OD 02-0 and 20α-hydroxyprogesterone (20α-OHP), also produced dose-dependent suppression of Ca(2+) influx, but had no effect on proliferation. Finally, inhibition of PHA-activated T-cell proliferation by progesterone is further suppressed by 20α-OHP, but not by Org OD 02-0. Overall, progesterone and R5020 are able to rapidly decrease PHA-stimulated sustained Ca(2+) influx, probably via blockade of TRPC3 channels, which suppresses T-cell proliferation. Taken together, the roles of progesterone and its analogs regarding the rapid response Ca(2+) influx need to be further explored in relation to cytokine secretion and proliferation in activated T-cells.

  19. CD4+ T cells dominate the leukocyte infiltration response initiated by intra-dermal injection of phytohemagglutinin into growing feathers in chickens.

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    Sullivan, K S; Erf, G F

    2017-10-01

    Phytohemagglutinin (PHA) is commonly used to evaluate cell-mediated immunocompetence. In chickens, PHA is typically injected intra-dermally (i.d.) into the skin (e.g., wing web, wattle, or footpad), and the tissue swelling response is monitored, whereby the extent of tissue swelling positively relates to the individual's cell-mediated immune system capabilities. Although i.d. injected PHA was shown to stimulate mononuclear cell and basophil infiltration to the site of injection, reports on temporal, qualitative, and quantitative aspects of the local cutaneous PHA response are limited. The objective of this study was to use the growing feather (GF) as a cutaneous test site to assess and monitor the type and relative amounts of leukocytes present in the pulp of PHA-injected GF. For this study, male, non-vaccinated Light-brown Leghorn chickens reared at the Arkansas Experiment Station Poultry Health Laboratory were used. At 9 wk of age, the dermis of 20 18-day-old regenerating GF was injected with 10 μL of either PBS diluent or 300 μg/mL PHA-P (5 chickens per treatment). GF were collected from each chicken before (zero) and at 0.25, 1, 2, 3, 4, 5, and 7 d post injection. At each time point, one GF was collected for immunofluorescent staining of pulp cell suspensions and leukocyte population analysis by flow cytometry, and another GF for histological analysis. Histological analysis confirmed participation of granulocytes and mononuclear cells, primarily lymphocytes, in the cutaneous PHA response. As revealed by flow cytometric cell population analysis, T cells, especially CD4+ T cells, constituted the major portion of the mononuclear cell infiltrate. Levels of CD4+ T cells were greatly elevated in PHA-injected GF within 6 h and remained elevated throughout the 7-day examination period. γδ T cells, CD8+ T cells, and B cells also infiltrated in response to PHA although at lower levels and with different time-course patterns from CD4+ T cells. The dominant presence

  20. Functional phytohemagglutinin (PHA) and Galanthus nivalis agglutinin (GNA) expressed in Pichia pastoris correct N-terminal processing and secretion of heterologous proteins expressed using the PHA-E signal peptide.

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    Raemaekers, R J; de Muro, L; Gatehouse, J A; Fordham-Skelton, A P

    1999-10-01

    Phytohemagglutinin (Phaseolus vulgaris agglutinin; PHA; E- and L-forms) and snowdrop lectin (Galanthus nivalis agglutinin; GNA) were expressed in Pichia pastoris using native signal peptides, or the Saccharomyces alpha-factor preprosequence, to direct proteins into the secretory pathway. PHA and GNA were present as soluble, functional proteins in culture supernatants when expressed from constructs containing the alpha-factor preprosequence. The recombinant lectins, purified by affinity chromatography, agglutinated rabbit erythrocytes at concentrations similar to the respective native lectins. However, incomplete processing of the signal sequence resulted in PHA-E, PHA-L and GNA with heterogenous N-termini, with the majority of the protein containing N-terminal extensions derived from the alpha-factor prosequence. Polypeptides in which most of the alpha-factor prosequence was present were also glycosylated. Inclusion of Glu-Ala repeats at the C-terminal end of the alpha-factor preprosequence led to efficient processing N-terminal to the Glu-Ala sequence, but inefficient removal of the repeats themselves, resulting in polypeptides with heterogenous N-termini still containing N-terminal extensions. In contrast, PHA expressed with the native signal peptide was secreted, correctly processed, and also fully functional. No expression of GNA from a construct containing the native GNA signal peptide was observed. The PHA-E signal peptide directed correct processing and secretion of both GNA and green fluorescent protein (GFP) when used in expression constructs, and is suggested to have general utility for synthesis of correctly processed proteins in Pichia.

  1. Differential expression of migration inhibitory and migration stimulatory factors in two lines of mice genetically selected for high or low responsiveness to phytohemagglutinin. 2. Effects of mitogenic or allogeneic stimulation.

    Science.gov (United States)

    Gauthier-Rahman, S; Suzuki, K; Couderc, J; el-Gharbi, N; Decreusefond, C; Stiffel, C

    1991-01-01

    Expression of migration inhibition factor (MIF) following in vitro stimulation with phytohemagglutinin (PHA) or allogeneic cells was explored in two lines of mice genetically selected for the high (Hi/PHA) or low (Lo/PHA) response of their lymphoid cells to PHA. Hi/PHA mice also have greater cell-mediated immune responses in mixed lymphocyte culture and graft-versus-host reactions, the poorer cell-mediated immune response of Lo/PHA being accompanied by a higher frequency of malignant tumours. Expression of MIF in PHA-pulsed spleen cell supernatants measured by a sensitive photoelectric method was found to be modulated by the concomitant presence of migration stimulation factor (MStF) derived from T cells. Both lymphokines were better expressed in Lo/PHA, as compared to Hi/PHA, under appropriate experimental conditions. Use of a low proliferative dose of mitogen (5 micrograms/ml PHA, 2-hour pulse) followed by culture in serum-free medium led Lo/PHA to express the highest titres of MIF, whereas a proliferative dose of PHA (50 micrograms/ml, 2-hour pulse) caused abrogation or occultation of expression of MIF and elective expression of MStF in this line. Hi/PHA mice expressed MIF equally at both mitogen doses, with transient expression of MStF followed by MIF after 50 micrograms/ml PHA, the kinetics of expression of the two lymphokines being different. Expression of MStF by spleen cells was an early event after PHA stimulation. In contrast to mitogenic stimulation, allogeneic stimulation in one-way mixed lymphocyte culture led to similar expression of MIF by both lines of mice. The implications of these findings are discussed.

  2. Differential expression of migration inhibitory and migration stimulatory factors in two lines of mice genetically selected for high or low responsiveness to phytohemagglutinin. 1. Migration stimulatory factor(s) from T and B cells of immune spleen.

    Science.gov (United States)

    Gauthier-Rahman, S; el-Gharbi, N; Siddiqui, M U; Couderc, J; Decreusefond, C; Stiffel, C

    1991-01-01

    Expression of the lymphokine migration inhibition factor in two lines of mice genetically selected for the high (Hi/PHA) or low (Lo/PHA) response of their lymph node cells to phytohemagglutinin was found to be modulated by concomitant expression of migration stimulation factor(s) [MStF(s)]. The expression of both lymphokines was dependent on genetic character and the immunizing dose of antigen. In mice immunized 5 days earlier with 50 micrograms ovalbumin in Freund's complete adjuvant (Ova in FCA immune), migration inhibition factor, assessed with a sensitive photoelectric method, was well expressed by male spleen or lymph node 24-hour culture supernatants of Lo/PHA but Hi/PHA, especially female, expressed marked MStF(s) instead. Immunization with 500 micrograms Ova in FCA markedly enhanced expression of MStF(s) in Lo/PHA but inhibited it in Hi/PHA. MStF(s) of Ova in FCA immune spleens of the two lines were found to derive from both T and B cells, B cell activity being greater. Lo/PHA were by far better expressors of both T- and B-cell-derived MStF(s) as compared to Hi/PHA (p less than 0.01). Spleen cells of mice immunized with FCA alone also expressed MStF(s) but to lesser extent than Ova in FCA immune spleens, expression by Lo/PHA B cells being significantly higher than in Hi/PHA (p less than 0.05). The MStF(s) of Ova in FCA immune spleens was found to be non-immunoglobulin in nature.

  3. 在日粮中添加葡萄糖氧化酶和植物血凝素防治仔猪早期断奶腹泻症的试验%Control of Piglet Early Weaning Diarrhea by Adding Glucose Oxidase and Phytohemagglutinin to Diet

    Institute of Scientific and Technical Information of China (English)

    田东霞; 张玉坤; 田泉成

    2012-01-01

    [目的]研究在日粮中添加葡萄糖氧化酶和植物血凝素防治仔猪早期断奶腹泻症的效果,探讨其对仔猪生产性能和肠道健康等方面所产生的作用。[方法]选用120头断奶仔猪,随机分为对照组、试验组I和试验组Ⅱ3组,分别饲喂常规断奶仔猪基础日粮、添加0.5%的葡萄糖氧化酶日粮以及添加0.5%的葡萄糖氧化酶和0.1%植物血凝素日粮,采用统一的饲养管理模式。记录每日腹泻次数、采食量、死亡数以及试验开始和结束时每头仔猪空腹体重:计算平均日增重、腹泻率、平均日采食量、饲料转化率及死亡率。[结果]试验期内仔猪平均日增重,试验组I和试验组Ⅱ分别比对照组提高25.0%和26.8%.差异极显著(P〈0.01);仔猪料肉比,试验组I和试验组Ⅱ分别比对照组下降17.2%和18.7%,差异板显著(P〈0.01);仔猪腹泻率,试验组I和试验组Ⅱ分别比对照组下降56.0%和65.5%,差异极显著(P〈0.01);仔猪成活率,试验组I比对照组提高7.4%,差异不显著(P〉0.05),试验组Ⅱ比对照组提高13%,差异极显著(P〈0.01)。[结论]在日粮中添加葡萄糖氧化酶和植物血凝素可明显提高饲料转化效率和仔猪成活卒.明显降低仔猪腹泻率。%[Objective] The control effect of piglet early weaning diarrhea (PEWD) by adding glucose oxidase and phytohemagglutinin to diet was studied to explore the effect on performance and intestinal health of piglet. [Method] One hundred and twenty weaned piglets were randomly divided into three groups, that was controlled, group I and group 11. Under the uniform raising management pattern, piglets in control were fed on routine basal diet; piglets in group I were fed on diet with 0.5% glucose oxidase; piglets in group II were fed on diet with 0.5% glucose oxidase and 0.1% phytohemagglutinin. The diarrhea frequency

  4. HIV-induced immunodeficiency. Relatively preserved phytohemagglutinin as opposed to decreased pokeweed mitogen responses may be due to possibly preserved responses via CD2/phytohemagglutinin pathway

    DEFF Research Database (Denmark)

    Hofmann, B; Jakobsen, K D; Odum, N

    1989-01-01

    We studied the proliferative response of PBL to the mitogens PHA and PWM and Candida albicans Ag in 301 HIV seropositive homosexual men, of whom 55 had AIDS. The responses to PHA were reduced only in the clinically ill HIV seropositive subjects. In contrast, the responses to PWM were profoundly...... and eight controls were chosen for the following studies. Expression of T3, Ti, delta receptors, and CD2 was investigated and showed an increased percentage of CD2 receptors positive cells in HIV seropositive subjects without AIDS. The proliferative responses of PBL to stimulation with PHA, PWM, antibodies...... to CD3, or antibodies to CD2 were investigated and showed significant correlation in controls, whereas in contrast, only the responses to PHA and CD2ab correlated in patients with AIDS. The proliferative responses to CD2ab and CD3ab in controls were larger than the responses to both PHA and PWM...

  5. [Genetic selection of mice for quantitative responsiveness of lymphocytes to phytohemagglutinin].

    Science.gov (United States)

    Stiffel, C; Liacopoulos-Briot, M; Decreusefond, C; Lambert, F

    1977-01-01

    A two-way selection was performed in mice according to the quantitative response of small lymphocytes to the mitogenic activity of phytohaemagglutinin (PHA). The response of inguinal lymph node cells of each mouse to an optimal dose of PHA was measured by 3H-thymidine incorporation using a micro-plate method. Starting from four outbred mouse strains we mated on the one hand mice getting the best response and on the other hand mice getting the poorest response. A progressive separation of the two lines was observed. At the 7th generation a 3-fold difference was found between the two lines. A similar interline difference was observed when concanavalin A (ConA) was used as mitogen. The separation of the two lines was also evident when spleen cells or thymus cells were cultured with PHA or ConA.

  6. A combined phytohemagglutinin and a-ketoglutarate pharmacology study of gut morphology and growth in older adult rats

    DEFF Research Database (Denmark)

    Filip, R.; Harrison, Adrian Paul; Pierzynowski, S.G.

    2008-01-01

    This study has evaluated the effect of phytohaemagglutinin (PHA) in combination with alpha-ketoglutaric acid (AKG), on GI-tract morphology and N balance in adult rats. Rats, aged approx. 15 months, were assigned to one of four experimental groups, (1) Control group, (2) AKG group, (3) AKG+PHA 100...

  7. Dietary supplementation with phytohemagglutinin in combination with a-ketoglutarate limits the excretion of nitrogen via urinary tract

    DEFF Research Database (Denmark)

    Filip, Rafal; Wdowiak, Leszek; Harrison, Adrian Paul;

    2008-01-01

    +PHA group of Controls; however, there was no significant difference in the thickness of the tunica mucosa. In the AKG+PHA group, the expression of neuropeptide Y (NPY) in the granula of neuronal cells of the submucosal parasympathetic ganglia was noticeable, although no expression was found in goblet cells...

  8. Structure and Activity Changes of Phytohemagglutinin from Red Kidney Bean (Phaseolus vulgaris) Affected by Ultrahigh-Pressure Treatments.

    Science.gov (United States)

    Lu, Yunjun; Liu, Cencen; Zhao, Mouming; Cui, Chun; Ren, Jiaoyan

    2015-11-04

    Phytohemagglutin (PHA), purified from red kidney beans (Phaseolus vulgaris) by Affi-Gel blue affinity chromatography, was subjected to ultrahigh-pressure (UHP) treatment (150, 250, 350, and 450 MPa). The purified PHA lost its hemagglutination activity after 450 MPa treatment and showed less pressure tolerance than crude PHA. However, the saccharide specificity and α-glucosidase inhibition activity of the purified PHA did not change much after UHP treatment. Electrophoresis staining by periodic acid-Schiff (PAS) manifested that the glycone structure of purified PHA remained stable even after 450 MPa pressure treatment. However, electrophoresis staining by Coomassie Blue as well as circular dichroism (CD) and differential scanning calorimetry (DSC) assay proved that the protein unit structure of purified PHA unfolded when treated at 0-250 MPa but reaggregates at 250-450 MPa. Therefore, the hemagglutination activity tends to be affected by the protein unit structure, while the stability of the glycone structure contributed to the remaining α-glucosidase inhibition activity.

  9. Dietary supplementation with phytohemagglutinin in combination with a-ketoglutarate limits the excretion of nitrogen via urinary tract

    DEFF Research Database (Denmark)

    Filip, Rafal; Wdowiak, Leszek; Harrison, Adrian Paul

    2008-01-01

    The aim of the study was to evaluate the effect of both phytohaemagglutinin (PHA) alone, and in combination with alpha-ketoglutaric acid (AKG), on nitrogen elimination via the urinary tract as opposed to the gastrointestinal tract of rats. In experiment 1, rats were assigned to one of two...... experimental groups, (1) Control and (2) PHA, whilst in experiment 2, rats were assigned to one of three experimental groups, (1) Control, (2) AKG, and (3) AKG+PHA. AKG was administered via drinking water, while PHA was administered via a stomach tube. The stock solution of crude PHA in 0.9% NaCl, was (20% w....../v) in water: 50 mg PHA/ml, 20 ml/kg body wt. Rats were 7 weeks old at the start of the experiments. Significantly lower daily weight gains in the AKG+PHA and PHA groups (pPHA...

  10. Dietary supplementation with phytohemagglutinin in combination with alpha-ketoglutarate limits the excretion of nitrogen via urinary tract

    DEFF Research Database (Denmark)

    Filip, Rafał; Wdowiak, Leszek; Harrison, Adrian P

    2008-01-01

    experimental groups, (1) Control and (2) PHA, whilst in experiment 2, rats were assigned to one of three experimental groups, (1) Control, (2) AKG, and (3) AKG+PHA. AKG was administered via drinking water, while PHA was administered via a stomach tube. The stock solution of crude PHA in 0.9 % NaCl, was (20 % w...

  11. Delayed hypersensitivity and migration inhibition in two lines of mice genetically selected for high or low responsiveness to phytohemagglutinin.

    Science.gov (United States)

    Gauthier-Rahman, S; El Rouby, S; Liacopoulos-Briot, M; Stiffel, C; Decreusefond, C; Liacopoulos, P

    1983-04-15

    Delayed-type hypersensitivity (DTH) and cell migration inhibition (MI) were studied in two lines of mice genetically selected for the high (Hi/PHA) or low (Lo/PHA) in vitro response of their lymphoid cells to phytochemagglutinin (PHA). A rapid photoelectric procedure for reading cell migrations enabled the study of MI over a wide range (10 log) of antigen concentrations in vitro. Hi/PHA mice required immunization with a 10 times higher dose of ovalbumin (OVA) in Freund's complete adjuvant (FCA) than Lo/PHA mice for a comparable response in DTH (footpad swelling) and MI of their induced peritoneal exudate cells (PEC). Lo/PHA spleen showed marked bizonal MI on Day 5 after immunization with low doses (0.1 and 0.5 micrograms) of OVA in FCA, one peak being obtained in presence of in vitro concentrations of 10(-3) or 10(-2) micrograms/ml OVA and another peak at 1 or 10 micrograms/ml, whereas Hi/PHA spleen showed stimulation of migration. In contrast, MI in Lo/PHA spleen failed to persist beyond Day 19, whereas it appeared progressively in Hi/PHA spleen, being maximal by Day 27. Low-zone inhibition in Hi/PHA spleen and PEC was lacking or poor even after immunization with higher doses of OVA in FCA. The implications of these findings are discussed.

  12. In vitro viability of lymphoid cells from lines of mice genetically selected for high or low responsiveness to phytohemagglutinin.

    Science.gov (United States)

    Stiffel, C; Liacopoulos-Briot, M; Decreusefond, C; Lambert, F

    1983-04-01

    The kinetics of viability of lymph node and spleen cells of mice genetically selected for "high" or "low" in vitro lymphocyte responsiveness to PHA were studied in PHA or PPD-stimulated short-term cultures. Lo/PHA cells were found to be less viable than Hi/PHA cells in unstimulated control cultures. PHA improved the viability of Lo/PHA cells while inducing proliferation of Hi/PHA cells with the appearance of more and larger lymphoblasts in the latter. PPD only improved the viability of spleen cell cultures, more so for the Hi/PHA line. The interline difference in thymidine uptake was smaller after PPD than after PHA stimulation. Modifications of culture conditions designed to decrease the interline difference in cell viability lessened but did not abolish the separation between the two lines for the PHA response as measured by thymidine uptake.

  13. EFFECTS OF β-ENDORPHIN ON PHYTOHEMAGGLUTININ -INDUCED LYMPHOCYTE PROLIFERATION AND MOUSE PLAQUE-FORMINGCELL RESPONSE VIA AN OPIOID RECEPTOR MECHANISM

    Institute of Scientific and Technical Information of China (English)

    林嘉友; 鲁刚; 翁佳玉

    1994-01-01

    The effects of opioid peptides on immune responses were investigated.It was found that β-endorphin(β-END) can depress proliferative responses to PHA in rat splenocytes but enhance those in mice,and it could also inibit the plaque-forming cell(PFC) response to sheep red blood cells when mouse splenocytes immunized in vivo were cultured in vitro with the peptide.The peptide antagonist naloxone was able to reverse β-END suppression of the PFC response.The data indicate that β-END suppresses antibody production or secretion via a specific opioidreceptor-mediated mechanism.

  14. Mixed lymphocyte reaction in mice genetically selected for high (Hi/PHA) or low (Lo/PHA) responsiveness to phytohemagglutinin. [Gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Liacopoulos-Briot, M. (Inst. d' Immuno-Biologie, Paris, France); Stiffel, C.; Lambert, F.; Decreusefond, C.

    1979-04-01

    Lymph node cells from Hi/PHA and Lo/PHA mice were evaluated for proliferative response after stimulation by allogeneic lymphocytes (MLR) originating from four inbred strains of different H-2 haplotype (C57B1/6, DBA/2, CBA, A). Reactivity to MLR and PHA were compared in these two lines and in the four inbred strains. The high and low responder status of Hi/PHA and Lo/PHA, as determined by T mitogens lymphocyte responsiveness, was also observed when one measured T responsiveness after MLR. Values obtained with the four inbred strains are included in the range of those measured in Hi/PHA and Lo/PHA cells when stimulated by PHA as well as by allogeneic cells. In contrast, when used as stimulator cells, Hi/PHA or Lo/PHA lymphocytes induce an equivalent proliferative response versus every responder inbred strain studied. These experiments support the hypothesis of a common genetic control of proliferative response following PHA or MLR stimulation. The genes implicated would be different from those coding for 1 region associated antigens.

  15. Protein nutritional quality of cowpea and navy bean residue fractions ...

    African Journals Online (AJOL)

    and navy bean residue-wheat diets was determined using in-vivo and in-vitro protein ... Phytohemagglutinin activity was only detectable in the raw cowpea ... Legume residues after protein extraction could be recommended for human food if

  16. Cellular Antigens in the Structure of Venezuelan Equine Encephalomyelitis Virus

    Science.gov (United States)

    1975-02-28

    emulsified in a physiological solution; 2) a mixture of tween (5 mg/ml of viral sus- pension) with two units of ether (20 min’s at 200 with agitation). The j...phytohemagglutinins. Ve employed th( mei:hod of sele:c:ive adsorption of antibodies of normal human serum a and ,3 (cor- respondingly lecithin ) according to

  17. Differences in dexamethasone-sensitivity between lymphocytes from patients with Alzheimer's disease and patients with multi-infarct dementia

    NARCIS (Netherlands)

    Nijhuis, E.W.P.; Oostervink, F.; Hinloopen, B.; Rozing, J.; Nagelkerken, L.

    1996-01-01

    Peripheral blood mononuclear cells (PBMC) from 40 consecutive patients entering a screening program on cognitive impairment were studied in vitro with respect to their sensitivity to dexamethasone (DEX). Phytohemagglutinin-induced proliferation by PBMC from patients with senile dementia of the Alzhe

  18. Granulocyte colony-stimulating factor increases CD4+ T cell counts of human immunodeficiency virus-infected patients receiving stable, highly active antiretroviral therapy

    DEFF Research Database (Denmark)

    Aladdin, H; Ullum, H; Dam Nielsen, S.

    2000-01-01

    counts resulted from increases in CD45RO+ memory T cells and cells expressing the CD38 activation marker. Lymphocyte proliferative responses to phytohemagglutinin and Candida antigen decreased, whereas NK cell activity and plasma HIV RNA did not change during G-CSF treatment. After 24 weeks, all immune......Thirty human immunodeficiency virus (HIV)-infected patients with CD4+ T cell counts

  19. Flow cytometric measurement of RNA synthesis based on bromouridine labelling and combined with measurement of DNA content or cell surface antigen

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J; Larsen, J K

    1993-01-01

    that RNA synthesis increased within the first 24 hours of phytohemagglutinin (PHA) stimulation, reaching a maximum at 48 hours, when cells had entered the cell cycle. Using a new method for flow cytometric dual parameter analysis of BrUrd incorporation and a cell surface antigen, spontaneous RNA synthesis...

  20. AcEST: DK961357 [AcEST

    Lifescience Database Archive (English)

    Full Text Available phytohemagglutinin OS=Ph... 56 2e-07 sp|P83410|LEC_ERYCG Lectin OS=Erythrina crista-galli PE=1 SV=1 56 2e-07...ragments) OS=Vicia villosa PE=1 ... 52 3e-06 sp|P16404|LEC_ERYCO Lectin OS=Erythrina corallodendron PE=1 SV=

  1. [Production of interleukin-2 by peripheral blood lymphocytes from patients with soft tissue sarcomas].

    Science.gov (United States)

    Berezhnaia, N M; Goretskiĭ, B A; Konovalenko, V F; Palivets, A Iu; Tolstopiatov, B A

    1987-01-01

    Interleukin-2 (IL-2) production of phytohemagglutinin-stimulated peripheral blood lymphocytes (PBL) was studied in 9 healthy subjects and 19 patients with soft tissue sarcomas. Mean IL-2 production by PBL in 19 patients was significantly diminished as compared with the control. Surgery leads to an increase of IL-2 production, however, the levels observed in the control do not restore completely.

  2. Inhibition of human lymphocyte proliferation and cleavage of interleukin-2 by Pseudomonas aeruginosa proteases

    DEFF Research Database (Denmark)

    Theander, T G; Kharazmi, A; Pedersen, B K

    1988-01-01

    This study was undertaken to determine the effect of Pseudomonas aeruginosa alkaline protease (AP) and elastase (ELA) on human lymphocyte function. AP at 50 micrograms/ml and ELA at 12 micrograms/ml caused a 50% inhibition of phytohemagglutinin-induced proliferation. There was no difference......, the inhibition was partly reversed. ELA at 10 micrograms/ml cleaved IL-2, as judged by size chromatography of a reaction mixture containing 125I-labeled IL-2 and the proteases. The ELA-digested IL-2 exhibited a reduced binding capacity to IL-2 receptors on the lymphocytes. Furthermore, treatment...... of phytohemagglutinin-stimulated lymphocytes with AP and ELA resulted in inhibition of binding of intact IL-2 to IL-2 receptors on the stimulated lymphocytes. These results indicated that P. aeruginosa-derived enzymes are able to interfere with human lymphocyte function in vitro and that this effect might be due...

  3. Studying the proliferation of human peripheral blood T lymphocytes in serum-free medium.

    Science.gov (United States)

    Tabakov, V U; Litvina, M M; Schepkina, J V; Jarilin, A A; Chestkov, V V

    2009-01-01

    We compared the cultivation of human peripheral blood lymphocytes in serum-free medium Hybris-2 and RPMI 1640 medium with 10% fetal bovine serum in the presence of phytohemagglutinin and interleukin-2. The optimal concentration of phytohemagglutinin significantly differed in serum-free and serum-containing media (0.5 and 5 microg/ml, [corrected] respectively). Both mitogens were more potent in stimulating the proliferation of lymphocytes in serum-free medium than in serum-containing medium. Strong proliferation of CD3(+) and CD4(+) T lymphocytes was observed in both media. The dynamics of other markers was similar in serum-free and serum-containing media. However, significant differences were revealed between individual donors. Our results indicate that the developed serum-free medium may be used in lymphocyte cultivation for scientific, diagnostic, and therapeutic purposes.

  4. Immune response and disease resistance of calves fed chromium nicotinic acid complex or chromium chloride.

    Science.gov (United States)

    Kegley, E B; Spears, J W; Brown, T T

    1996-07-01

    Twenty-one Holstein bull calves (skinfold thickness after an intradermal injection of phytohemagglutinin were measured to evaluate cell-mediated immune response. Calves supplemented with Cr-nicotinic acid complex had a greater response than did controls at 6, 12, 24, and 48 h after injection. Calves supplemented with CrCl3 had a greater response than did controls at 24 and 48 h after injection. In vitro blastogenic responses of lymphocytes to phytohemagglutinin or pokeweed mitogen and antibody response to porcine red blood cells were not affected by treatment. Following a disease challenge with an intranasal dose of infectious bovine rhinotracheitis on d 75, body temperature tended to be lower for calves supplemented with Cr-nicotinic acid complex than for control calves. Calves supplemented with either Cr source had lower serum cortisol concentrations at 5 d after challenge. Chromium supplementation enhanced cell-mediated immune function.

  5. Influence of Phthalates on in vitro Innate and Adaptive Immune Responses

    DEFF Research Database (Denmark)

    Hansen, Juliana Frohnert; Nielsen, Claus Henrik; Brorson, Marianne Møller

    2015-01-01

    Phthalates are a group of endocrine disrupting chemicals, suspected to influence the immune system. The aim of this study was to investigate the influence of phthalates on cytokine secretion from human peripheral blood mononuclear cells. Escherichia coli lipopolysaccharide and phytohemagglutinin-...... not seem to be a result of cell death. Thus, results indicate that both human innate and adaptive immunity is influenced in vitro by phthalates, and that phthalates therefore may affect cell differentiation and regenerative and inflammatory processes in vivo.......Phthalates are a group of endocrine disrupting chemicals, suspected to influence the immune system. The aim of this study was to investigate the influence of phthalates on cytokine secretion from human peripheral blood mononuclear cells. Escherichia coli lipopolysaccharide and phytohemagglutinin...

  6. Porcine leukocyte cellular subsets sensitive to African swine fever virus in vitro.

    OpenAIRE

    1984-01-01

    African swine fever virus infected most, if not all, of the macrophages (monocytes) and ca. 4% of the polymorphonuclear leukocytes from porcine peripheral blood. B and T lymphocytes, either resting or stimulated with phytohemagglutinin, lipopolysaccharide, or pokeweed mitogen, were not susceptible to the virus. All of the mitogens used inhibited African swine fever multiplication in susceptible cells. The number of virus passages in vitro and the virulence degree of the virus did not affect t...

  7. Selective Regulation of Human Immunodeficiency Virus-Infected CD4+ Lymphocytes by a Synthetic Immunomodulator Leads to Potent Virus Suppression In Vitro and in hu-PBL-SCID Mice

    OpenAIRE

    Bahr, George M.; Darcissac, Edith C. A.; Castéran, Nathalie; Amiel, Corinne; Cocude, Cécile; Truong, Marie-José; Dewulf, Joëlle; Capron, André; Mouton, Yves

    2001-01-01

    We have previously observed that the synthetic immunomodulator Murabutide inhibits human immunodeficiency virus type 1 (HIV-1) replication at multiple levels in macrophages and dendritic cells. The present study was designed to profile the activity of Murabutide on CD8-depleted phytohemagglutinin-activated lymphocytes from HIV-1-infected subjects and on the outcome of HIV-1 infection in severe combined immunodeficiency mice reconstituted with human peripheral blood leukocytes (hu-PBL-SCID mic...

  8. Cellular immune findings in Lyme disease.

    Science.gov (United States)

    Sigal, L. H.; Moffat, C. M.; Steere, A. C.; Dwyer, J. M.

    1984-01-01

    From 1981 through 1983, we did the first testing of cellular immunity in Lyme disease. Active established Lyme disease was often associated with lymphopenia, less spontaneous suppressor cell activity than normal, and a heightened response of lymphocytes to phytohemagglutinin and Lyme spirochetal antigens. Thus, a major feature of the immune response during active disease seems to be a lessening of suppression, but it is not yet known whether this response plays a role in the pathophysiology of the disease. PMID:6240164

  9. Ageing and cell-mediated immunity.

    Science.gov (United States)

    Fixa, B; Komárková, O; Chmelar, V

    1975-01-01

    The lymphocyte transformation test with phytohemagglutinin as mitogen estimated according to the incorporation of 2-(14)C-thymidine in DNA was used as an indicator of cell-mediated reactivity in 53 healthy subjects. Three age groups were examined: up to 20 years (21 subjects), 21-40 years (10 subjects) and over 70 years (22 subjects). The responsiveness of lymphocytes decreased significantly with age. In the highest age group 12 pathologically low values were found.

  10. Links between CD147 Function, Glycosylation, and Caveolin-1

    OpenAIRE

    Tang, Wei; Chang, Sharon B.; Hemler, Martin E.

    2004-01-01

    Cell surface CD147 shows remarkable variations in size (31-65 kDa) because of heterogeneous N-glycosylation, with the most highly glycosylated forms functioning to induce matrix metalloproteinase (MMP) production. Here we show that all three CD147 N-glycosylation sites make similar contributions to both high and low glycoforms (HG- and LG-CD147). l-Phytohemagglutinin lectin binding and swainsonine inhibition experiments indicated that HG-CD147 contains N-acetylglucosaminyltransferase V-cataly...

  11. Suppressor cell hyperactivity relative to allogeneic lymphocyte proliferation as a manifestation of defective T-T-cell interactions in systemic lupus erythematosus

    Energy Technology Data Exchange (ETDEWEB)

    Stenina, M.A.; Potapova, A.A.; Biryukov, A.V.; Skripnik, A.Yu.; Cheredeev, A.N.

    1987-01-01

    The authors study the state of immunoregulatory process in patients with systemic lupus erythematosus at the T-T-cell interaction level and seek to test the possibility of the pharmacological modulation of this process. The proliferative activity of mononuclear lymphocytes, extracted from the blood of ten lupus patients, was assessed by measuring the incorporation of tritiated thymidine into cultures stimulated by phytohemagglutinin, concanavalin, and theophylline. The comparative effects of each of these agents on the immunoregulatory and proliferative activity of the lymphocytes are reported.

  12. Cytotoxicity of lymphocytes from melanoma patients against autologous tumor cells and its potentiation in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Bykovskaya, S.N.; Iobadze, M.S.; Kupriyanova, T.A.; Demidov, L.V.

    1987-06-01

    The specific and natural cytotoxicity of peripheral blood lymphocytes from patients with melanomas was compared and stimulation with autologous tumor cells or a pool of allogeneic lymphocytes from five healthy blood donors also was used to potentiate the specific antitumor activity of the patients' lymphocytes. To assess cytolytic ability, cells of an autologous tumor, cells of the K-562 line, autologous peripheral blood lymphocytes, and blast cells obtained from these lymphocytes after stimulation by phytohemagglutinin were used as the target cells. The target cells were incubated in a medium containing sodium chromate and were labelled with the chromium 51 isotope.

  13. Reactivating effect of levamisole on cell-mediated immunity in gastrointestinal cancer patients

    Directory of Open Access Journals (Sweden)

    Miwa,Hiroaki

    1977-10-01

    Full Text Available Cell-mediated immunity was studied in 23 cases of advanced gastrointestinal cancer. The patients received levamisole at 150 mg/day for three consecutive days each week for four weeks. In cases at the terminal stage of gastrointestinal cancer, the blastformation rate of peripheral blood lymphocytes against phytohemagglutinin (PHA after the administration of levamisole showed a slight increase, but cases with blastformation rates over 40% increased markedly three or four weeks after the initial administration of levamisole. The peripheral blood lymphocyte count showed little change in these cases.

  14. Analysis of common bean expressed sequence tags identifies sulfur metabolic pathways active in seed and sulfur-rich proteins highly expressed in the absence of phaseolin and major lectins

    Science.gov (United States)

    2011-01-01

    Background A deficiency in phaseolin and phytohemagglutinin is associated with a near doubling of sulfur amino acid content in genetically related lines of common bean (Phaseolus vulgaris), particularly cysteine, elevated by 70%, and methionine, elevated by 10%. This mostly takes place at the expense of an abundant non-protein amino acid, S-methyl-cysteine. The deficiency in phaseolin and phytohemagglutinin is mainly compensated by increased levels of the 11S globulin legumin and residual lectins. Legumin, albumin-2, defensin and albumin-1 were previously identified as contributing to the increased sulfur amino acid content in the mutant line, on the basis of similarity to proteins from other legumes. Results Profiling of free amino acid in developing seeds of the BAT93 reference genotype revealed a biphasic accumulation of gamma-glutamyl-S-methyl-cysteine, the main soluble form of S-methyl-cysteine, with a lag phase occurring during storage protein accumulation. A collection of 30,147 expressed sequence tags (ESTs) was generated from four developmental stages, corresponding to distinct phases of gamma-glutamyl-S-methyl-cysteine accumulation, and covering the transitions to reserve accumulation and dessication. Analysis of gene ontology categories indicated the occurrence of multiple sulfur metabolic pathways, including all enzymatic activities responsible for sulfate assimilation, de novo cysteine and methionine biosynthesis. Integration of genomic and proteomic data enabled the identification and isolation of cDNAs coding for legumin, albumin-2, defensin D1 and albumin-1A and -B induced in the absence of phaseolin and phytohemagglutinin. Their deduced amino acid sequences have a higher content of cysteine than methionine, providing an explanation for the preferential increase of cysteine in the mutant line. Conclusion The EST collection provides a foundation to further investigate sulfur metabolism and the differential accumulation of sulfur amino acids in seed

  15. Analysis of common bean expressed sequence tags identifies sulfur metabolic pathways active in seed and sulfur-rich proteins highly expressed in the absence of phaseolin and major lectins

    Directory of Open Access Journals (Sweden)

    Sharpe Andrew

    2011-05-01

    Full Text Available Abstract Background A deficiency in phaseolin and phytohemagglutinin is associated with a near doubling of sulfur amino acid content in genetically related lines of common bean (Phaseolus vulgaris, particularly cysteine, elevated by 70%, and methionine, elevated by 10%. This mostly takes place at the expense of an abundant non-protein amino acid, S-methyl-cysteine. The deficiency in phaseolin and phytohemagglutinin is mainly compensated by increased levels of the 11S globulin legumin and residual lectins. Legumin, albumin-2, defensin and albumin-1 were previously identified as contributing to the increased sulfur amino acid content in the mutant line, on the basis of similarity to proteins from other legumes. Results Profiling of free amino acid in developing seeds of the BAT93 reference genotype revealed a biphasic accumulation of gamma-glutamyl-S-methyl-cysteine, the main soluble form of S-methyl-cysteine, with a lag phase occurring during storage protein accumulation. A collection of 30,147 expressed sequence tags (ESTs was generated from four developmental stages, corresponding to distinct phases of gamma-glutamyl-S-methyl-cysteine accumulation, and covering the transitions to reserve accumulation and dessication. Analysis of gene ontology categories indicated the occurrence of multiple sulfur metabolic pathways, including all enzymatic activities responsible for sulfate assimilation, de novo cysteine and methionine biosynthesis. Integration of genomic and proteomic data enabled the identification and isolation of cDNAs coding for legumin, albumin-2, defensin D1 and albumin-1A and -B induced in the absence of phaseolin and phytohemagglutinin. Their deduced amino acid sequences have a higher content of cysteine than methionine, providing an explanation for the preferential increase of cysteine in the mutant line. Conclusion The EST collection provides a foundation to further investigate sulfur metabolism and the differential accumulation of

  16. Cytogenetic studies in twelve patients with primary myelofibrosis and myeloid metaplasia.

    Science.gov (United States)

    Smadja, N; Krulik, M; de Gramont, A; Sirinelli, A; Brissaud, P; Dray, C; Audebert, A A; Debray, J

    1987-01-01

    Chromosome studies on bone marrow and/or peripheral blood cells without phytohemagglutinin were performed on 12 patients with primary myelofibrosis with myeloid meta-plasia (PMMM) between 1980 and 1984. Abnormal clones were found in six patients (50%). In five cases the abnormal clone involved the long arm of chromosome #7, two of which also had partial trisomy of chromosome #1 and trisomy of 9. Additional abnormalities involving chromosomes #3, #5, #11, #13, #15, and #21 were each found once. Review of the literature showed few studies on the cytogenetics of PMMM. No specific chromosomal pattern can be established; however, abnormalities described are nonrandom.

  17. Quantification of thymidine kinase (TK1) mRNA in normal and leukemic cells and investigation of structure-function relatiosnhip of recombinant TK1enzyme

    DEFF Research Database (Denmark)

    Kristensen, Tina

    patients with chronic lymphatic leukemia (CLL). 2: Structure-function relationship of recombinant TKI. In the first part a sensitive method (competitive PCR) for quantification of TKI mRNA was established. The TKI mRNA level was quantified in quiescent lymphocytes from control donors (n = 6......) and in lymphocytes stimulated to growth by the mitogen phytohemagglutinin. The expression in normal cells was compared with the level of TK1 mRNA level in patients with chronic lymphatic leukemia (n = 5). The results for the six control donors show a very low level of TK1 mRNA (below 0.006~1 O6 copies mg-’ protein...

  18. Flow cytometric analysis of p21 protein expression on irradiated human lymphocytes; Analise por citometria de fluxo da expressao da proteina p21 em linfocitos humanos irradiados

    Energy Technology Data Exchange (ETDEWEB)

    Santos, N.F.G.; Amaral, A., E-mail: neyliane@gmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Laboratorio de Modelagem e Biodosimetria Aplicada; Freitas-Silva, R. [Universidade Federal de Pernambuco (UFPE), Garanhuns, PE (Brazil). Departamento de Ciencias Naturais e Exatas; Pereira, V.R.A. [Fundacao Oswaldo Cruz (FIOCRUZ), Recife, PE (Brazil). Centro de Pesquisas Aggeu Magalhaes. Departamento de Imunologia. Lab. de Imunoparasitologia; Tasat, D.R. [Universidad Nacional de General San Martin, Buenos Aires (Argentina). Escuela de Ciencia y Tecnologia. Laboratorio de Biologia Celular del Pulmon

    2013-08-15

    Cell cycle blockage in G1 is a mechanism p21 protein-regulated and coupled to DNA damage response to permit genetic content analysis, damage repair and cell death. Analysis of proteins that participates of this response has progressed with new analytic tools, and data contributes to comprehension of radioinduced molecular events as well as to new approaches on practices that employ ionizing radiation. On this perspective, the aim of this research was to evaluate, by flow cytometry, p21 expression on irradiated human lymphocytes, maintained under different experimental conditions. Peripheral blood samples from 10 healthy subjects were irradiated with doses of 0 (non-irradiated), 1, 2 and 4 Gy. Lymphocytes were processed to analysis on ex vivo (no cultured) condition and after 24; 48 and 72 hours culture, with and without phytohemagglutinin stimulation. p21 protein expression levels were measured by flow cytometry, as percentage values. Results indicate that flow cytometric assay allows detection of changes on p21 expression, since it was detected significant increase on phytohemagglutinin-stimulated samples, for all times, against basal expression (ex vivo). However, it was not observed significant alterations on p21 protein radioinduced levels, for all doses, times and culture conditions analyzed. These results not indicate so p21 protein as bioindicator of ionizing radiation exposure. Nevertheless, data confirmation may to require analysis of a more numerous population. (author)

  19. Use of whole blood lymphocyte stimulation test for immunocompetency studies in bald eagles, red-tailed hawks, and great horned owls.

    Science.gov (United States)

    Redig, P T; Dunnette, J L; Sivanandan, V

    1984-11-01

    Mitogen-induced whole blood lymphocyte stimulation tests for immunocompetency studies in bald eagles (Haliaeetus leucocephalus), red-tailed hawks (Buteo jamaicensis), and great horned owls (Bubo virginianus) were developed. Combinations of incubation times, blood dilutions, concentrations of [3H]thymidine and [125I]2-deoxyuridine, antibiotics, phytohemagglutinin-P, and concanavalin A were tested for their effects on the stimulation index (SI). An antibiotic combination of gentamicin plus amphotericin B yielded low SI with lymphocytes from bald eagles, but not with lymphocytes from great horned owls or red-tailed hawks. Penicillin plus streptomycin caused no such depression of SI. Lymphocytes from all 3 species yielded maximum responses with a 48-hour prelabel and 12- to- 16 hour postlabel incubation period at 41 C and 1:20 blood dilution. Optimal mitogen concentrations for lymphocytes from bald eagles, red-tailed hawks, and great horned owls were 25 micrograms, 10 micrograms, and 10 micrograms of phytohemagglutinin-P/well, respectively, and 2.5 micrograms, 10 micrograms, and 10 micrograms of concanavalin A/well, respectively. Differences in SI were not seen between the 2 radioactive labels. The optimal concentration of the [3H]thymidine label ranged from 0.06 to 0.125 microCi/well.

  20. Infectious Bursal Disease: Pathogenicity and Immunogenicity of Vaccines

    Directory of Open Access Journals (Sweden)

    E Camilotti

    Full Text Available ABSTRACT The Infectious Bursal Disease (IBD is a contagious viral disease that affects young chickens and may cause high morbidity and mortality. As the virus is very resistant to the environment, vaccination is required in case of high infection pressure. Due to variations in the virulence degree of the vaccines available to control IBD, this study aimed at evaluating the pathogenicity and immunogenicity of three types of vaccines. In total, 220 one-day-old specific pathogen free (SPF chickens were immunized with recombinant, immune-complex and intermediate vaccines, or not vaccinated (55 birds per group and challenged with IBD G11 strain on day 25. On days 25, 30, and 35, the Bursa of Fabricius (BF were submitted to gross and histological examination, and serum samples were submitted to ELISA to determined anti-IBD antibody titers. On day 23, chickens were submitted to the test of hypersensitivity to phytohemagglutinin to evaluate the immunosuppressive effect of vaccines on the cell-mediated immunity. The results have indicated that the immune-complex vaccine induced the most severe BF lesions, whereas the recombinant vaccine preserved BF tissue and cell integrity. The three evaluated vaccines induced humoral immunity of similar intensity. The cellular reaction to phytohemagglutinin of the chickens immunized with recombinant and immune-complex vaccines was less severe compared with the unvaccinated chickens. In conclusion, these results indicate that the immune-complex vaccine was the most pathogenic and that all vaccines were effective in protecting SPF chickens against IBD.

  1. Individual susceptibility to DNA telomerase inhibitors: a study on the chromosome instability induced by 3'-azido-3'-deoxythymidine in lymphocytes of elderly twins.

    Science.gov (United States)

    Caporossi, Daniela; Argentin, Gabriella; Pittaluga, Monica; Parisi, Paolo; Tedeschi, Bruna; Vernole, Patrizia; Cicchetti, Rosadele

    2004-03-01

    The activation of telomerase in phytohemagglutinin-stimulated peripheral lymphocytes is thought to play a role in telomere maintenance and DNA repair. Considering the importance of this enzyme in both cancer and senescence, we studied the effects of the telomerase inhibitor 3'-azido-3'-deoxythymidine on the frequency of chromosomal aberrations and micronuclei induced in peripheral blood lymphocytes (PBL) of elderly monozygotic and dizygotic twins, evaluated with respect to the genotoxic effects induced in unrelated young subjects. Our results show that the cytogenetic damage induced by 3'-azido-3'-deoxythymidine in human PBL was mainly regulated by genetic factors and allowed the identification of hypersensitive subjects. Ageing, which did not modify the individual susceptibility to 3'-azido-3'-deoxythymidine induction of chromosome aberrations and micronuclei, nevertheless determined an overall increase in nuclear damage.

  2. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Science.gov (United States)

    Hill, F. S.; Cox, A. B.; Salmon, Y. L.; Cantu, A. O.; Lucas, J. N.

    1994-01-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 microgram/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) than that produced by PHA (MI < 0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture.

  3. The immune privilege of the eye: human retinal pigment epithelial cells selectively modulate T-cell activation in vitro

    DEFF Research Database (Denmark)

    Kaestel, Charlotte G; Lovato, Paola; Ødum, Niels

    2005-01-01

    PURPOSE: To examine the effect of human retinal pigment epithelial (RPE) cells on phytohemagglutinin (PHA) activation of T cells. METHODS: Resting peripheral blood lymphocytes (PBLs) were stimulated with PHA with or without the presence of gamma-irradiated RPE cells. Proliferation and the cell...... cycle profile were thereafter investigated by 3H-thymidine incorporation and flow cytometric analysis. In addition, the PBLs expression of CD69, major histocompatibility complex (MHC) class I and II, CD3, as well as the IL-2 receptor chains were evaluated by flow cytometry, and the content of IL-2...... in cell culture supernatant was measured by ELISA. RESULTS: Human RPE cells were found to suppress PHA-induced proliferation, cyclin A, IL-2R-alpha and -gamma, and CD71 expression and decrease the production of IL-2; but RPE cells do not inhibit the PHA-induced expression of early activation markers CD69...

  4. Increased production of interleukin-6 by T lymphocytes from patients with multiple myeloma.

    Science.gov (United States)

    Lapeña, P; Prieto, A; Garcia-Suarez, J; Reyes, E; San Miguel, J; Jorda, J; Alvarez-Mon, M

    1996-01-01

    Alterations in T lymphocyte functions may affect other cellular components of the immune system. Several lymphokines produced by T cells are involved in the proliferation and differentiation of human B lymphocytes. Alterations in the secretion of these molecules may be implicated in the development of B cell lymphoproliferative diseases. We have investigated the production of interleukin-2 (IL-2) and interleukin-6 (IL-6) by T lymphocytes from 14 patients with multiple myeloma (MM) and 16 healthy controls. The phenotypical and functional characteristics of these T lymphocytes were also studied. The proliferative response to vegetal lectin phytohemagglutinin (PHA) stimulation was decreased in T lymphocytes from MM patients (p 0.05) but not by exogenous IL-6 (p lectin stimulation, the production of IL-2 by T lymphocytes from those patients was normal, while IL-6 secretion was increased.

  5. Parathyroid hormone dependent T cell proliferation in uremic rats

    DEFF Research Database (Denmark)

    Lewin, E; Ladefoged, Jens; Brandi, L

    1993-01-01

    Chronic renal failure (CRF) is combined with an impairment of the immune system. The T cell may be a target for the action of parathyroid hormone (PTH). Rats with CRF have high blood levels of PTH. Therefore, the present investigation examined some aspects of the T cell function in both normal...... and CRF rats before and after parathyroidectomy and after an isogenic kidney transplantation. The T cell proliferative response to phytohemagglutinin (PHA) stimulation was significantly higher in peripheral blood mononuclear cell (PBMC) cultures obtained from CRF rats than from normal rats. After...... parathyroidectomy the T cells of normal as well as of uremic rats could still be significantly stimulated by PHA, but now no significant difference was seen. When CRF was reversed after an isogenic kidney transplantation and PTH reversed to levels in the normal range, the T cell proliferative response to PHA...

  6. Clinical study of non-specific cell mediated immunity in the patients with esophageal cancer. Influence of preoperative irradiation and surgical intervention

    Energy Technology Data Exchange (ETDEWEB)

    Nakata, Yoshitaka

    1987-06-01

    Few data are available to elucidate the influence of combined preoperative irradiation and surgery on the non-specific cell mediated immunity of patients with esophageal cancer. In vitro and in vivo examinations of the non-specific cell mediated immunity were made before and after irradiation and surgery in 108 patients with esophageal cancer. Decreased immune competence was noticeable one month after surgery in the irradiated group, as compared with the non-irradiated group. Simultaneously, the ratio of concanavalin A to phytohemagglutinin was significantly higher in the irradiated group than the non-irradiated group (p < 0.01). Two months later, both findings in the two groups were similar. There was no consistent tendency toward altered immune competence between the group with curative surgery and the group with non-curative surgery. (Namekawa, K.).

  7. Alteration of T cell function in healthy persons with a history of thymic x irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Rieger, C.H.L.; Kraft, S.C.; Rothberg, R.M.

    1975-10-01

    The possible late effects of x irradiation to the infantile thymus were investigated by studying immune functions in 12 healthy persons with a history of thymic x irradiation and healthy control subjects. No differences were found in serum immunoglobulin values, humoral antibody levels, lymphocyte counts, and lymphocyte reactivity to phytohemagglutinin, vaccinia virus, purified protein derivative (PPD), and allogeneic cells. The irradiation group exhibited cellular hyperresponsiveness to streptokinase-streptodornase (SK-SD). In contrast, mean skin and in vitro lymphocyte responses to Candida albicans were depressed in the patients with thymic irradiation. A dissociation of these two Candida responses was found in only 1 of 14 healthy control subjects but in 7 of 12 irradiated individuals. While thymic irradiation did not result in impaired immunologic defenses leading to clinical disease, it caused alterations in T cell responses similar to those reported in patients with chronic mucocutaneous candidiasis.

  8. Immunological aspects of the common food colorants, amaranth and tartrazine.

    Science.gov (United States)

    Koutsogeorgopoulou, L; Maravelias, C; Methenitou, G; Koutselinis, A

    1998-02-01

    We describe a sensitive and reproducible microassay model using human peripheral blood lymphocytes (PBL) for discrimination between the cytotoxic and immunosuppressive effects of food colorants such as amaranth and tartrazine. The cytotoxic effects of a wide range of concentrations of these substances were studied on human PBL by the colorimetric in vitro cytotoxicity assays, neutral red uptake (NR) and thiazolyl blue tetrazolium bromide (MTT). The immunotoxic properties of these 2 substances were determined by a [3H]-thymidine DNA incorporation assay on phytohemagglutinin stimulated or non-stimulated lymphocytes, as well as by a Cr51 release Natural Killer assays. The results showed clear immunosuppressive effects from the 2 substances tested, although the concentrations chosen for this study proved to be non-cytotoxic by NR and MTT cytotoxic endpoints.

  9. Lymphocyte transformation in presumed ocular histoplasmosis

    Energy Technology Data Exchange (ETDEWEB)

    Ganley, J.P.; Nemo, G.J.; Comstock, G.W.; Brody, J.A.

    1981-08-01

    Lymphocytes from individuals with inactive macular disciform lesions of presumed ocular histoplasmosis challenged with three histoplasmin antigens incorporated tritiated thymidine at a significantly higher rate than histoplasmin-stimulated lymphocytes of matched control and peripheral scar groups. This finding is consistent with the etiologic association of the disciform ocular syndrome and previous systemic infection with Histoplasma capsulatum. The disciform group had a higher mean response than the other two groups to pokeweed mitogen but not to phytohemagglutinin and had higher mean counts per minute to the specific antigens Toxoplasma gondii, Blastomyces dermatitidis, Cryptococcus neoformans, Mycobacterium tuberculosis, M battery, and M gaus, but not to Candida albicans. These data would suggest that individuals with the disciform lesion of presumed ocular histoplasmosis have a hyperreactive cellular immune response; this response may play an important role in the development of the disciform.

  10. Suppressed peripheral blood lymphocyte blastogenesis in pre- and postpartal sheep by chronic heat-stress, and suppressive property of heat-stressed sheep serum on lymphocytes.

    Science.gov (United States)

    Niwano, Y; Becker, B A; Mitra, R; Caldwell, C W; Abdalla, E B; Johnson, H D

    1990-01-01

    Phytohemagglutinin (PHA) and concanavalin A (Con A)-induced blastogenesis of peripheral blood lymphocytes was examined in heat-stressed pre- and postpartal sheep. The peak responses of lymphocytes to PHA and Con A in heat-stressed sheep revealed significant reduction before and after parturition compared with those in the corresponding control animals kept under thermoneutral conditions. Furthermore, the effect of serum from control or heat-stressed sheep on PHA-induced lymphocyte blastogenesis was examined. Supplementation of serum from heat-stressed sheep significantly suppressed the blastogenesis of lymphocytes obtained from healthy sheep, bovine, and human donors. Unlike dexamethasone, heat-stressed sheep serum did not inhibit IL-2 production by PHA-stimulated human peripheral blood lymphocytes. These results indicate that the immunosuppression of heat-stressed sheep is in part mediated by serum factor(s) that can modulate T-cell function in a species nonspecific manner.

  11. Habitat structure is associated with the expression of carotenoid-based coloration in nestling blue tits Parus caeruleus

    Science.gov (United States)

    Arriero, Elena; Fargallo, Juan Antonio

    2006-04-01

    We investigated how the expression of carotenoid-based plumage coloration (lightness and chroma) in nestling blue tits Parus caeruleus is associated with forest structure in oak forests of central Spain. We found evidence of a reduced expression of carotenoid-based coloration in nestlings growing up in successionally young and structurally simple forest territories. Our results suggest that breast feather coloration can be used as an indicator of nestling quality because nestlings with more intense yellow plumage coloration had larger body size and stronger immune responses to the injection of phytohemagglutinin (PHA). Given the association of forest structural complexity with carotenoid-based plumage coloration, our findings suggest that variation in habitat structure may have a significant impact on forest birds in their first stages of life which has implications for forest management practices.

  12. Multiple dysfunctions in developmental and activational stages of T lymphocytes, B lymphocytes and monocytes in ARC and AIDS patients.

    Science.gov (United States)

    Sei, Y; Tsang, P H; Petrella, R J; Bekesi, J G

    1987-11-01

    Peripheral blood leukocytes from ARC and AIDS patients were examined before and after phytohemagglutinin (PHA) stimulation by dual color flow cytometry and monoclonal antibodies which identify developmental and activational stages of T lymphocytes, B cells and monocytes. There was a persistent elevation in the total number of circulating Ia+ lymphocytes with progressive selection for B1+ Ia+ lymphocytes and T suppressor cells and a concurrent reduction in the antigen-presenting monocytes. Following PHA stimulation there was a marked decrease in all subsets of Ia+ lymphocytes and monocytes. These results indicate (a) multicellular dysfunctions in the immunosurveillance mechanisms in AIDS, and (b) that many functional subsets of circulating lymphocytes and monocytes were already activated and therefore poorly responsive to additional antigenic or mitogenic stimuli.

  13. In Vitro Interleukin-1 and 2 Production and Interleukin 2 Receptor Expression in the Rhesus Monkey

    Science.gov (United States)

    Schmitt, Didier A.; Sonnenfeld, Gerald; Husson, David; Tkaczuk, Jean; Andre, Eric; Schaffar, Laurance

    1996-01-01

    Anti-human monoclonal antibodies were used to detect and quantify interleukins-1 and 2 and interleukin-2 receptor expression in peripheral blood mononuclear cells from a rhesus monkey. Interleukin-1 production could be induced by phorbol esters (PMA) and was potentiated by phytohemagglutinin (PHA). Interleukin-2 secretion could also be induced by the combination of PHA and PMA, but only weakly with PHA alone. Interleukin-2 receptor expression was present in a subpopulation of unstimulated lymphocytes and could be enhanced by PHA or PMA. These data show once again that the rhesus monkey immune system is cross-reactive with the human one and that rhesus macaque could be a good model to study interleukin therapy.

  14. In vitro capacity of various cyclooxygenase inhibitors to revert immune suppression caused by radiation therapy for breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Blomgren, H.; Rotstein, S. (Karolinska Sjukhuset, Stockholm (Sweden)); Wasserman, J.; Petrini, B. (Stockholm County Council (Sweden). Central Microbiological Laboratory); Hammarstroem, S. (Linkoeping University Faculty of Health Sciences (Sweden). Department of Cell Biology, Division of Medical and Physical Chemistry)

    1990-12-01

    Radiation therapy triggers blood monocytes to an increased secretion of immunosuppressive prostaglandins (PGs), which in part can explain the post-irradiation impairment of lymphocyte blastogenesis. Since low mitogen responses of lymphocytes in irradiated breast cancer patients is linked to a poor prognosis a clinical trial is planned to examine if treatment with inhibitors of PG-synthesis during irradiation can counteract immunosuppression and increase survival. In the present investigation the authors have compared 9 different inhibitors of PG-synthesis for capacity to enhance phytohemagglutinin responses of blood lymphocytes before and after irradiation for breast cancer. 5 of the drugs (aspisol, indomethacin, meclofenamic acid, ketoprofen and diclofenac) enhanced the reactivity to more than 150 percent. In general, the strongest enhancements were observed in lymphocyte preparations obtained at completion of irradiation when reactivity was most depressed followed by those obtained at one month and before irradiation. (author). 28 refs.; 5 figs.; 1 tab.

  15. Differential chromosomal radiosensitivity within the first G1-phase of the cell cycle of early-dividing human leukocytes in vitro after stimulation with PHA

    Energy Technology Data Exchange (ETDEWEB)

    Beek, B.; Obe, G.

    1977-02-11

    Human leukocyte cultures were irradiated with 200 R X rays before the addition of phytohemagglutinin (PHA) in the Go-stage and at different times up to 25 h within the first G1-phase of the cell cycle after the addition of PHA. The results of the analysis of chromosomal aberrations show that the frequencies of dicentric chromosomes increase significantly when leukocytes leave the Go-stage, reaching a minimum yield of aberrations about halfway through the first G1-phase. After that, toward the end of the G1-phase, the frequencies of dicentric chromosomes decrease again to a level similar to that found in the Go-stage. Different possible explanations for the differential chromosomal radiosensitivity of human leukocytes within the first post-stimulation G1-phase are discussed.

  16. The effect of ceramic fibers on the immune system.

    Science.gov (United States)

    Tulinská, Jana; Kuricová, Miroslava; Lisková, Aurelia; Kováciková, Zuzana; Tatrai, Elizabeth

    2005-12-01

    Male Sprague-Dawley rats were treated by intratracheal instillation with 1 mg/animal of refractory ceramic fibers. Intratracheal exposure to ceramic fibers led to significant changes of immune response. Results of proliferative activity of spleen lymphocytes showed significantly decreased proliferative activity of T-cells in response to mitogens phytohemagglutinin and concanavalin A in animals given ceramic fibers in comparison with control rats. Similarly, T-dependent B-cell response to pokeweed mitogen was significantly suppressed. Spontaneous proliferative activity of lymphocytes in non-stimulated spleen cell cultures did not differ in exposed and control rats. No significant changes were found among groups in percentage of phagocytic blood polymorphonuclear leukocytes and percentage of cells with respiratory burst.

  17. Effect of gomisin A in the prevention of acute hepatic failure induction.

    Science.gov (United States)

    Mizoguchi, Y; Kawada, N; Ichikawa, Y; Tsutsui, H

    1991-08-01

    Nearly all rats develop massive hepatic cell necrosis and die upon intravenous administration of heat-killed Propionibacterium acnes followed by a small amount of Gram-negative lipopolysaccharide 7 days later. However, when such an experimental liver disorder is induced in rats raised for 4 or more weeks on food containing 0.06% of gomisin A extracted and purified from Schizandra chinensis, the survival rate rises, histological changes of the liver improve remarkably, and splenocyte reactivity to phytohemagglutinin and pokeweed mitogen as well as splenocyte interleukin 1 productivity are retained. These results suggested the possibility that the development of acute hepatic failure may be prevented with the oral administration of gomisin A.

  18. Functional and phenotypic changes in human lymphocytes after coincubation with Leishmania donovani in vitro

    DEFF Research Database (Denmark)

    Hviid, L; Sørensen, A L; Kharazmi, A

    1990-01-01

    that the inhibition of the proliferative response to PHA by live L. donovani in vitro is associated with early processes in lymphocyte activation. Further studies on the inhibitory phenomena described may be of potential significance in the investigation of the suppressive mechanisms in human visceral leishmaniasis.......In this paper we describe functional and phenotypic changes in T cells after in vitro coincubation of peripheral blood mononuclear cells (PBMC) and Leishmania donovani parasites at different parasite/peripheral blood mononuclear cell ratios. The phytohemagglutinin (PHA)-induced lymphoproliferative...... response was reduced by the coincubation, and at the maximal parasite/peripheral blood mononuclear cell ratio used (7.5:1), the average response was less than 40% of the response in the absence of parasites. The cause of the reduction in lymphoproliferation is not clear, but it requires live parasites...

  19. Specific in vivo and nonspecific in vitro alloreactivities of adult frogs (Xenopus laevis) that were thymectomized during early larval life.

    Science.gov (United States)

    Nagata, S; Cohen, N

    1983-07-01

    Thymectomy of very young Xenopus larvae abrogated in vitro proliferative responses to phytohemagglutinin (PHA) and allogeneic leukocytes. Thymectomized (Txd) frogs, however, still rejected first-set skin allografts chronically and second-set grafts (but not third-party grafts) more rapidly. The specific second-set rejection reaction by Txd frogs was transferable in vivo to secondary Txd hosts (with major histocompatibility complex identical to the cell donor) by the subcutaneous injection of a mixture of splenic and peripheral blood leukocytes. Spleen cells used in this adoptive transfer experiment (i.e., immune cells) were responsive to allogeneic cells from the original donor strain as well as from unrelated donors in mixed leukocyte culture in vitro but were still unreactive to PHA. The results are consistent with an extrathymic pathway of alloreactive cell differentiation in this species, although the physiological significance of such a pathway is not clear.

  20. Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas.

    Science.gov (United States)

    Muñoz, Fernando A; Estrada-Parra, Sergio; Romero-Rojas, Andres; Work, Thierry M; Gonzalez-Ballesteros, Erik; Estrada-Garcia, Iris

    2009-10-15

    To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6k Da band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demonstration of CD3+ cells in reptiles using specific antibodies.

  1. Lactobacilli Modulate Natural Killer Cell Responses In Vitro

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen; Christensen, Hanne Risager; Frøkiær, Hanne

    Natural killer (NK) cells are cells of the non-specific immune system lysing altered self-cells. A non-cytolytic subset of NK cells may serve a regulatory role by secreting cytokines. Bacteria translocating across the gastrointestinal mucosa are presumed to gain access to NK cells, as consumption...... of certain lactic acid bacteria has been shown to increase in vivo NK cytotoxicity. Here, we investigated how human gut flora-derived lactobacilli affect NK cells in vitro, by measuring proliferation and IFN-gamma production of human NK cells upon bacterial stimulation. CD3-CD56+ NK cells were isolated from...... buffy coats by negative isolation using non-NK lineage specific antibodies and magnetic beads. NK cells were incubated with 10 microg/ml UV-inactivated bacteria or 10 microg/ml phytohemagglutinin (PHA) for four days. Proliferation was assessed by incorporation of radioactive thymidine into NK cell DNA...

  2. Reduced LAK cytotoxicity of peripheral blood mononuclear cells in patients with bladder cancer

    DEFF Research Database (Denmark)

    Hermann, G G; Petersen, K R; Steven, K

    1990-01-01

    were analyzed using monoclonal antibodies against T cells, natural killer (NK) -cells, monocytes, and activation markers. The cytotoxicities of US-PBMC, PS-PBMC, and LAK cells were all significantly lower in the cancer patients than in the controls (P less than 0.05). The percentages of PBMC positive......The cytotoxicity of unstimulated peripheral blood mononuclear cells (US-PBMC), phytohemagglutinin (PHA)-stimulated PBMC (PS-PBMC) and interleukin-2 (IL-2)-activated PBMC (LAK cells) was assessed in patients with noninvasive and invasive transitional-cell bladder cancer and compared with those...... determined in healthy controls. The differences in the cytotoxicities were correlated with specific changes in the subsets of peripheral blood mononuclear cells (PBMC). PBMC from 37 patients and 13 healthy controls were tested against the bladder cancer cell line T24 in 51Cr-release assays. The PBMC subsets...

  3. Natural Killer Cells Are Activated by Lactic Acid Bacteria-Matured Dendritic Cells

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen; Christensen, Hanne Risager; Frøkiær, Hanne

    Natural killer (NK) cells are cells of the non-specific immune system lysing altered self-cells. A non-cytolytic subset of NK cells may serve a regulatory role by secreting cytokines. Bacteria translocating across the gastrointestinal mucosa are presumed to gain access to NK cells, as consumption...... of certain lactic acid bacteria has been shown to increase in vivo NK cytotoxicity. Here, we investigated how human gut flora-derived lactobacilli affect NK cells in vitro, by measuring proliferation and IFN-gamma production of human NK cells upon bacterial stimulation. Human peripheral blood NK cells were...... incubated with 10 microg/ml UV-inactivated bacteria or 10 microg/ml phytohemagglutinin (PHA) for four days. Proliferation was assessed by incorporation of radioactive thymidine into NK cell DNA. The IFN-gamma concentration was measured by ELISA. Incubation of NK cells with a Lactobacillus acidophilus strain...

  4. Cellular immunity in Hodgkin's disease.

    Science.gov (United States)

    Advani, S H; D'Silva, H; Gothoskar, B P; Dinshaw, K A; Nair, C N; Gopalkrishna, R; Talwalkar, G V; Desai, P B

    1979-02-01

    Defective cell-mediated immunity (CMI) occurs early in the course of Hodgkin's disease (HD) and may persist even two years after successful treatment. This has been confirmed by in vivo and in vitro tests performed on 51 untreated and 52 treated patients of HD. The grading of skin reponse in vivo to dinitrochlorobenzene (DNCB) correlated very well with the in vitro leukocyte migration inhibition (LMI) response against phytohemagglutinin (PHA). An inhibitory influence of HD patients' sera was demonstrated by LMI tests in vitro. The response of peripheral leukocytes from HD patients in the LMI tests could be augmented in vitro by addition of levamisole (an immuno-potentiator) to the culture medium, thus pointing to an intrinsic defect in Lymphocytes. The data indicate that defect at multiple sites in the immune system is responsible for persistent anergy in HD.

  5. Estrogenic xenobiotics affect the intracellular activation signal in mitogen-induced human peripheral blood lymphocytes: immunotoxicological impact.

    Science.gov (United States)

    Sakabe, K; Okuma, M; Kazuno, M; Yamaguchi, T; Yoshida, T; Furuya, H; Kayama, F; Suwa, Y; Fujii, W; Fresa, K L

    1998-01-01

    The present study was an attempt to elucidate the effect of estrogenic xenobiotics on the proliferation of mitogen-stimulated human peripheral blood lymphocyte (PBL). Our findings follow: (a) the proliferation of PBL in response to phytohemagglutinin (PHA) was mediated by protein kinase C activity, but estrogenic xenobiotics had a strong inhibitory effect on protein kinase C activity of PHA-stimulated PBL; (b) cytoplasmic extracts from PHA-stimulated PBL greatly activated DNA replication, but estrogenic xenobiotics had a strong inhibitory effect on these activities. The results suggest that the cytoplasmic signal-generating system in mitogen-treated PBL is inhibited by estrogenic xenobiotics, and that the defect occurs at all stages in the sequence of events leading to DNA synthesis and cell proliferation.

  6. Genotoxic effect of formocresol pulp therapy of deciduous teeth.

    Science.gov (United States)

    Lucas Leite, Ana Catarina Gaioso; Rosenblatt, Aronita; da Silva Calixto, Merilane; da Silva, Cirlene Maria; Santos, Neide

    2012-08-30

    To investigate whether formocresol, in Buckley's original formulation, used for pulp therapy of deciduous teeth, can have a genotoxic effect. Genotoxicity was tested in lymphocyte cultures from the peripheral blood of children aged 5-10y, in Recife, Pernambuco, Brazil. This was a case-control study. The sample comprised 40 children who had primary teeth with non-vital pulps. Two venous blood samples (6-8ml) were collected from each child, the first prior to pulp therapy (control group) and the second 24h after pulp therapy (experimental group). Lymphocyte cultures were grown in 78% RPMI 1640 medium, 20% fetal bovine serum, 2% phytohemagglutinin. The lymphocytes were assessed for chromosomal aberrations; each sample involved analysis of 100 metaphases. There was a statistically significant difference between the control and treated groups for the isochromatid gap (pformocresol in pediatric dentistry is recommended. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Tumor xenotransplantation in Wistar rats after treatment with cyclophosphamide and total lymphoid irradiation. [X-ray

    Energy Technology Data Exchange (ETDEWEB)

    Hoogenhout, J. (St. Radbond Academic Hospital, Nijmegen, Netherlands); Kazem, I.; Jerusalem, C.R.; Bakkeren, J.A.J.; de Jong, J.; Kal, H.B.; van Munster, P.J.J.

    1982-10-01

    Three-month-old male Wistar rats were treated with cyclophosphamide and total lymphoid irradiation, and C22LR mouse osteosarcoma was transplanted into the rats. The effects of immunosuppression were monitored by lymphocyte counts, serum IgG determinations, phytohemagglutinin (PHA) and concanavalin A (Con A) responses, measurement of the proportion of B cells, and histopathological studies of the lymphoid organs. At eight days after treatment, the lymphocyte counts, IgG levels, and PHA and Con A values were decreased. Mitotic activity started in the depleted B and T cell areas of the peripheral lymphatic organs two weeks after treatment. There was a 94% graft take of the osteosarcoma. It was determined that the optimum time for tumor xenograft transplantation is 4 days after treatment. The duration of growth was 11 days, and this was followed by regression up to day 21.

  8. Tumor xenotransplantation in Wistar rats after treatment with cyclophosphamide and total lymphoid irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Hoogenhout, J.; Kazem, I.; Jerusalem, C.R.; Bakkeren, J.A.; de Jong, J.; Kal, H.B.; van Munster, P.J.

    1982-10-01

    Three-month-old male Wistar rats were treated with cyclophosphamide and total lymphoid irradiation, and C22LR mouse osteosarcoma was transplanted into the rats. The effects of immunosuppression were monitored by lymphocyte counts, serum IgG determinations, phytohemagglutinin (PHA) and concanavalin A (Con A) responses, measurement of the proportion of B cells, and histopathological studies of the lymphoid organs. At eight days after treatment, the lymphocyte counts, IgG levels, and PHA and Con A values were decreased. Mitotic activity started in the depleted B and T cell areas of the peripheral lymphatic organs two weeks after treatment. There was a 94% graft take of the osteosarcoma. It was determined that the optimum time for tumor xenograft transplantation is 4 days after treatment. The duration of growth was 11 days, and this was followed by regression up to day 21.

  9. Potential for the G2/M arrest assay to predict patient susceptibility to severe reactions following radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Perez, A.; Grabenbauer, G.G.; Sauer, R.; Distel, L.V.R. [Dept. of Radiation Oncology, Friedrich Alexander Univ. Erlangen-Nuremberg (Germany); Sprung, C.N. [Div. of Research, Peter MacCallum Cancer Centre, and Dept. of Biochemistry and Molecular Biology, Melbourne Univ., VIC (Australia)

    2007-02-15

    Background and purpose: cell-cycle regulation and checkpoint activation are crucial factors for radiation-induced DNA damage processing. The G2/M phase arrest was assessed in lymphoblastoid cell lines and phytohemagglutinin-stimulated T-lymphocytes of different radiosensitivities to study the relationship of G2/M arrest to radiosensitivity. Material and methods: G2/M arrest was analyzed after in vitro irradiation by 2 and 5 Gy of ionizing radiation up to 6 days using 17 lymphoblastoid cell lines from healthy individuals, ataxia-telangiectasia (AT) patients, Nijmegen breakage syndrome (NBS) patients and cancer patients with clinically increased radiosensitivity. In a second approach, phytohemagglutinin-stimulated T-lymphocytes from 15 healthy individuals, twelve cancer patients, and five cancer patients hypersensitive to ionizing radiation were studied. Image cytometry was performed to analyze G2/M arrest. Results: two of the three AT cell lines showed markedly increased G2/M arrest compared to controls. NBS cells were comparable to controls up to day 3, but then demonstrated a slightly increased G2/M arrest. Two of the six radiosensitive lymphoblast cell lines and the five radiosensitive cancer patients' T-lymphocytes assayed showed a reduction in G2/M arrest, while healthy individuals showed no difference from cancer patients. Conclusion: the interrelation between G2/M arrest and radiosensitivity is not readily apparent since a variety of radiosensitive cells from patients with radiosensitive syndromes and patients identified as radiosensitive following radiation treatment showed inconsistent G2/M arrest dynamics. Secondary effects, like loss of clonogenicity, G1/S phase arrest and failure of G2/M arrest may contribute to variation of the G2/M arrest endpoint and obscure assessment of cellular radiosensitivity using this method. (orig.)

  10. Modest weight loss through a 12-week weight management program with behavioral modification seems to attenuate inflammatory responses in young obese Koreans.

    Science.gov (United States)

    Lee, AeJin; Jeon, Kyeong Jin; Kim, Min Soo; Kim, Hye-Kyeong; Han, Sung Nim

    2015-04-01

    Obesity has been reported to impair immune functions and lead to low-grade long-term inflammation; however, studies that have investigated the impact of weight loss on these among the young and slightly obese are limited. Thus, we investigated the effect of a 12-week weight management program with behavioral modifications on cell-mediated immune functions and inflammatory responses in young obese participants. Our hypothesis was that weight loss would result in improved immune functions and decreased inflammatory responses. Sixty-four participants (45 obese and 19 normal weight) finished the program. Obese (body mass index ≥25) participants took part in 5 group education and 6 individual counseling sessions. Normal-weight (body mass index 18.5-23) participants only attended 6 individual sessions. The goal for the obese was to lose 0.5 kg/wk by reducing their intake by 300 to 500 kcal/d and increasing their physical activity. Program participation resulted in a modest but significant decrease in weight (2.7 ± 0.4 kg, P < .001) and lipopolysaccharide-stimulated interleukin-1β production (from 0.85 ± 0.07 to 0.67 ± 0.07 ng/mL, P < .05) in the obese. In the obese group, increase in phytohemagglutinin-stimulated interleukin-10 production, a TH2 and anti-inflammatory cytokine, approached significance after program participation (from 6181 ± 475 to 6970 ± 632 pg/mL, P = .06). No significant changes in proliferative responses to the optimal concentration of concanavalin A or phytohemagglutinin were observed in the obese after program participation. Collectively, modest weight loss did not change the cell-mediated immune functions significantly but did attenuate the inflammatory response in young and otherwise healthy obese adults.

  11. Changes in cytokine production and composition of peripheral blood leukocytes during pregnancy are not associated with a difference in the proliferative immune response to the fetus.

    Science.gov (United States)

    Lashley, Lisa E E L O; van der Hoorn, Marie-Louise P; van der Mast, Barbara J; Tilburgs, Tamara; van der Lee, Nadine; van der Keur, Carin; van Beelen, Els; Roelen, Dave L; Claas, Frans H J; Scherjon, Sicco A

    2011-10-01

    We analyzed peripheral blood from women at term pregnancy for leukocyte composition, in vitro proliferative responses and cytokine production after nonspecific and fetus-specific stimulation. Maternal peripheral blood mononuclear cells (PBMCs) were collected and stimulated with umbilical cord blood (UCB) of the mother's own child, third-party UCB, nonspecific stimulus phytohemagglutinin, and anti-CD3 antibody, with PBMCs of nonpregnant women (cPBMC) as controls. Nine combinations of patient, child, third party child, and controls were selected on basis of sharing one human leukocyte antigen (HLA)-DR antigen. The response of mPBMC upon specific stimulation with fetal antigens was similar to that of cPBMC. No differences were found when comparing the mother's response upon stimulation to her own child with stimulation to that with a control child. Nonspecific stimulation with phytohemagglutinin and anti-CD3 antibody did not reveal a difference in proliferation rate between mPBMC and cPBMC. However, mPBMC contained a higher percentage of CD14(+) cells (p = 0.001) and activated T cells (CD25(dim), p < 0.0001), but a lower percentage CD16(-)CD56(bright) natural killer (NK) cells (p = 0.001) and CD16(+)CD56(+) NK cells (p = 0.003). mPBMC produced more interleukin (IL)-6, IL-10, and IL-17 compared with cPBMC (p < 0.05). We found differences in lymphocyte composition and cytokine production between mPBMC and cPBMC. These differences did not result in quantitative changes in proliferative responses during pregnancy compared with responses in nonpregnant controls. Copyright © 2011 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.

  12. Effects of increased human tumor necrosis factor-like molecule 1A expression in peripheral blood of children with acute Guillain-Barre syndrome on interferon-gamma secretion

    Institute of Scientific and Technical Information of China (English)

    Libin Yang; Shulei Li; Yan Tana; Shufen Xu; Xiumei Duan; Yanqiu Fang; Lihua Liu; Yuanyuan Che; Lei Liu

    2009-01-01

    BACKGROUND:Human tumor necrosis factor-like molecule 1A (hTL1A) is a strong T helper cell type 1 (Th1) co-stimulator.Guillain-Barre syndrome (GBS) is an autoimmune disorder of the nervous system,which is mediated by Th1 cells.OBJECTIVE:To determine hTL1A expression in peripheral blood T lymphocytes of acute GBS children and the effects of hTL1A on secretion of interferon-γ.DESIGN,TIME AND SETTING:A randomized,controlled,neuroimmunological in vitro study was performed at the Central Laboratory of First Hospital of Jilin University,China from November 2005 to November 2007.MATERIALS:Venous blood samples were obtained from 6 healthy donors,aged 6-12 years (all routine blood examination items were normal),and 6 additional children with acute GBS,aged 6-12years.The GBS children fell itl within 1 week and were not treated with hormones or immunoglobulin.Purified recombinant human soluble tumor necrosis factor-like molecule 1A (rhsTL1A,1 mg/mL,relative molecular mass 22 000,6×His tag,soluble form) was supplied by the Central Laboratory of First Hospital of Jilin University,China.METHODS:Peripheral blood mononuclear cells were isolated from healthy donors using the standard Ficoll gradient centrifugation and were incubated in 96-well culture plates.The cells were assigned to the following groups:control (2 μg/mL phytohemagglutinin),2 μg/mL phytohemagglutinin+25,100 and 400 ng/mL rhsTL1A.T cell proliferation was quantified using the tritiated thymidine (~3H-TdR) method.Serum interferon-γ levels in acute GBS children were detected by enzyme-linked immunosorbent assay (ELISA).The ratio of hTL1A-positive T cells to CD3-positive T cells in peripheral blood of acute GBS children was determined using flow cytometry.Following in vitro pre-activation of peripheral blood mononuclear cells by 2 μg/mL phytohemagglutinin,the peripheral blood mononuclear cells were treated with 400 ng/mL exogenous rhsTL1A.Finally,peripheral blood mononuclear cell-secreted interferon-γ levels were

  13. In vitro screening of plant lectins and tropical plant extracts for anthelmintic properties.

    Science.gov (United States)

    Ríos-de Álvarez, L; Jackson, F; Greer, A; Bartley, Y; Bartley, D J; Grant, G; Huntley, J F

    2012-05-25

    Lectins are plant secondary metabolites (PSM) found in many forages and which may confer anthelmintic properties to gastrointestinal parasites through disrupting the development of parasitic larvae throughout its life cycle. In experiment 1, the ability of the plant lectins jacalin (JAC), concanavalin A (Con A), phytohemagglutinin E2L2 (PHA-E2L2), phytohemagglutinin L4 (PHA-L4), phytohemagglutinin E3L (PHA-E3L), kidney bean albumin (KBA), Robinia pseudoacacia agglutinin (RPA), Maackia amurensis lectin (MAA), Maclura pomifera agglutinin (MAA), Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA) and Galanthus nivalis agglutinin (GNA) to disrupt the feeding of the first stage larvae (L(1)) of the sheep gastro-intestinal nematodes (GIN) Teladorsagia circumcincta, Haemonchus contortus and Trichostrongylus colubriformis was investigated using a larval feeding inhibition test (LFIT). Only PHA-E3L, WGA and Con A had a potent effect on disrupting larval feeding of all of the three species of GIN investigated. The lectin concentration required to inhibit feeding in 50% of L(1) (IC50) was 7.3±1.2, 8.3±1.4 and 4.3±1.7 μg/ml for PHA-E3L; 59.1±32.4, 58.7±11.9 and 8.1±7.0 μg/ml for Con A and 78.9±11.2, 69.4±8.1 and 28.0±14.1 μg/ml for WGA for T. circumcincta, H. contortus and T. colubriformis larvae, respectively (P=0.006). The addition of the lectin inhibitors fetuin, glucose/mannose or N-acetylglucosamine for PHA-E3L, Con A and WGA, respectively, caused an increase in the proportion of larvae that had fed at all concentrations for PHA-E3L only. In experiment 2, the effect of extracts from the tropical plants Azadiractha indica, Trichanthera gigantea, Morus alba, Gliricidia sepium and Leucaena leucocephala on the feeding behaviour of H. contortus L(1,) was examined. A. indica, T. gigantea and M. alba failed to inhibit 50% of larvae from feeding at concentrations up to 10mg plant extract per ml. In contrast, both G. sepium and L. leucocephala demonstrated

  14. 1. cap alpha. ,25-Dihydroxyvitamin D/sub 3/ inhibits. gamma. -interferon synthesis by normal human peripheral blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Reichel, H.; Koeffler, H.P.; Tobler, A.; Norman, A.W.

    1987-05-01

    1..cap alpha..,25-Dihydroxyvitamin D/sub 3/ (1,25-(OH)/sub 2/D/sub 3/), the biologically active metabolite of vitamin D/sub 3/, inhibited synthesis of ..gamma..-interferon (IFN-..gamma..) by phytohemagglutinin-activated peripheral blood lymphocytes (PBLs). A significant reduction of IFN-..gamma.. protein levels in PBL culture medium was achieved with a physiologic 1,25-(OH)/sub 2/D/sub 3/ concentration, 1,25-(OH)/sub 2/D/sub 3/ also inhibited accumulation of IFN-..gamma.. mRNA in activated PBLs in a dose-dependent fashion. The ability of 1,25-(OH)/sub 2/D/sub 3/ to modulate IFN-..gamma.. protein synthesis was unaltered in the presence of high concentrations of recombinant human interleukin 2. The suppression of IFN-..gamma.. synthesis by PBLs was specific for 1,25-(OH)/sub 2/D/sub 3/; the potencies of other vitamin D/sub 3/ metabolites were correlated with their affinities for the cellular 1,25-(OH)/sub 2/D/sub 3/ receptor. The time course of 1,25-(OH)/sub 2/D/sub 3/ receptor expression in phytohemagglutinin-activated PBLs was correlated with the time course of 1,25-(OH)/sub 2/D/sub 3/-mediated inhibition of IFN-..gamma.. synthesis. Finally, the authors examined the effects of 1,25-(OH)/sub 2/D/sub 3/ on the constitutive IFN-..gamma.. production by two human T-lymphocyte lines transformed by human T-lymphotropic virus type I. The cell lines were established from a normal donor (cell line S-LB1) and from a patient with vitamin D-dependent rickets type 2 (cell line Ab-VDR). IFN-..gamma.. synthesis by S-LB1 cells was inhibited in a dose-dependent fashion by 1,25-(OH)/sub 2/D/sub 3/, whereas IFN-..gamma.. synthesis by Ab-VDR cells was not altered by 1,25-(OH)/sub 2/D/sub 3/. The data presented in this study provide evidence for a role of 1,25-(OH)/sub 2/D/sub 3/ in immunoregulation.

  15. Leishmania donovani: assessment of leishmanicidal effects of herbal extracts obtained from plants in the visceral leishmaniasis endemic area of Bihar, India.

    Science.gov (United States)

    Singh, Shubhankar K; Bimal, Sanjiva; Narayan, Shyam; Jee, Chandrawati; Bimal, Devla; Das, P; Bimal, Raageeva

    2011-02-01

    One obstacle faced in the effective control of visceral leishmaniasis (VL) is the limited number of available treatment options. Furthermore, control efforts have been hindered further by the emergence of Leishmania resistance to many of the available drugs. In this study, we investigated the anti-leishmanial properties of 30 medicinally important plants from the VL endemic area of Bihar, India and compared them to two available anti-leishmanial drugs (sodium antimony gluconate and amphotericin B) and two plant lectins (phytohemagglutinin and concanavalin A) on Leishmania donovani promastigotes in vitro at 24 and 48 h after initiation of culture. We identified eight plant extracts in addition to phytohemagglutinin and amphotericin B that significantly inhibited the growth of promastigotes (p Agave americana, Azadirachta indica, Eclipta alba and Piper longum) of the eight plant extracts that induced significant promastigotes killing (p = 0.00098). Effect-based dose finding analysis revealed that the threshold concentration of A. americana required to eliminate L. donovani after 24h was 0.05 mg/ml. A. indica and P. longum plant extracts eliminated L. donovani promastigotes after 48 h at concentrations of 0.1 and 0.5mg/ml, respectively. E. alba eliminated the promastigotes at a concentration of 0.5mg/ml within 24h. The axenic amastigote killing response was 1.90-, 2.52- and 1.3-fold higher than the promastigote killing response with A. indica, A. americana and E. alba plant extracts, respectively. A. americana and A. indica, respectively, led to approximate 2.5- and 1.3-fold declines in mitochondrial dehydrogenase activity compared with control. E. alba stimulation resulted in an up-regulation of dehydrogenase activity (p = 0.00329). The CSA from P. longum was found to be least cytotoxic; the observed difference in mitochondrial activity was insignificant (p = 0.16314). Further studies may reveal the pharmacological significance of many of the plants with anti

  16. Human B cell activating factor (BCAF): production by a human T cell tumor line.

    Science.gov (United States)

    Fevrier, M; Diu, A; Mollier, P; Abadie, A; Olive, D; Mawas, C; Theze, J

    1989-01-01

    In a previous study, we demonstrated that supernatants from human T cell clones stimulated by a pair of anti-CD2 monoclonal antibodies cause resting human B cells to become activated and to proliferate in the absence of any other signals. The activity responsible for these effects was shown to be different from already characterized lymphokines and in particular from IL-2 and IL-4, and was named B Cell Activating Factor or BCAF. In this paper, we describe the production of BCAF by a human T cell tumor line T687 after phorbol myristate acetate (PMA) stimulation; this production can be potentiated by phytohemagglutinin (PHA). We further show that the stimulatory phase can be separated from the secretory phase thereby avoiding contamination of BCAF-containing supernatant by PMA and PHA. Supernatants produced under these conditions do not contain either IL-4 or IFN but contain traces of lymphotoxin and 2 to 10 ng/ml of IL-2. The T687 cell line will allow us to obtain a large volume of supernatant for biochemical study and purification of the molecule(s) responsible for BCAF activity.

  17. Enhancement of cellular and humoral immunity following embryonic exposure to melatonin in turkeys (Meleagris gallopavo).

    Science.gov (United States)

    Moore, C B; Siopes, T D

    2005-09-01

    Two experiments were performed to determine the effect of in ovo melatonin supplementation on the ontogeny of immunity in the Large White turkey poult. Different levels of melatonin were injected into the air cell of the egg 4 days prior to hatch. In Experiment 1, turkey embryos received 3 ml of solution containing 200, 100, 50, 25, 10, or 1 microg/ml of melatonin. The hatchability at each dose was determined and compared to vehicle-injected controls. In Experiment 2, only poults from melatonin treatments in Experiment 1 that resulted in normal hatchability (10 and 1 microg/ml) were used. Lymphoproliferative responses to phytohemagglutinin (PHA-P) and primary antibody responses to Chukar red blood cells (CRBC) were determine at five time intervals: 0, 1, 7, 14, and 21 days post-hatch. At each of these times, including 28 days post-hatch, treatment effects on body weights were determined. At 28 days post-hatch, bursal, thymic, and splenic weights were obtained. In ovo melatonin administration significantly accelerated (P0.05) the development of cell-mediated (PHA-P) and humoral (CRBC) immune responses, and these responses were significantly elevated above vehicle-injected controls through 21 days post-hatch. No effect was observed on bursal, thymic, splenic or body weights. These data suggest that embryonic exposure to melatonin enhances post-hatch immune development and responsiveness.

  18. Refolding and Characterization of Recombinant Human GST-PD-1 Fusion Protein Expressed in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    Da-Wei LI; Jian-Feng YU; Yong-Jing CHEN; Hong-Bing MA; Zheng-Fei WANG; Yi-Bei ZHU; Xue-Guang ZHANG

    2004-01-01

    Programmed death-1 (PD-1) is a costimulatory molecule of CD28 family expressed onactivated T, B and myeloid cells. The engagement of PD-1 with its two ligands, PD-L1 and PD-L2, inhibitsproliferation of T cell and production of a series of its cytokines. The blockade of PD-1 pathway is involvedin antiviral and antitumoral immunity. In this study, human PD-1 cDNA encoding extracellular domain wasamplified and cloned into expression plasmid pGEX-5x-3. The fusion protein GST-PD-1 was effectivelyexpressed in E. coli BL21 (DE3) as inclusion bodies and a denaturation and refolding procedure was performed to obtain bioactive soluble GST-PD-1. Fusion protein of above 95% purity was acquired by a convenient two-step purification using GST affinity and size exclusion columns. Furthermore, a PD-L1-dependentin vitro bioassay method was set up to characterize GST-PD-1 bioactivity. The results suggested that GSTPD-1 could competently block the interaction between PD-L1 and PD-1 and increase the production of IL2 and IFN-γ of phytohemagglutinin-activated T cells.

  19. Refoiding and Characterization of Recombinant Human GST-PD-1 Fusion Protein Expressed in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    Da-WeiLI; Jian-FengYU; Yong-JingCHEN; Hong-BingMA; Zheng-FeiWANG; Yi-BeiZHU; Xue-GuangZHANG

    2004-01-01

    Programmed death-1 (PD-1) is a costimulatory molecule of CD28 family expressed onactivated T, B and myeloid cells. The engagement of PD-1 with its two ligands, PD-L1 and PD-L2, inhibitsproliferation of T cell and production of a series of its cytokines. The blockade of PD-1 pathway is involvedin antiviral and antitumoral immunity. In this study, human PD-1 cDNA encoding extracellular domain wasamplified and cloned into expression plasmid pGEX-Sx-3. The fusion protein GST-PD-1 was effectivelyexpressed in E. coli BL21 (DE3) as inclusion bodies and a denaturation and refolding procedure was per-formed to obtain bioactive soluble GST-PD-I. Fusion protein of above 95% purity was acquired by a conve-nient two-step purification using GST affinity and size exclusion columns. Furthermore, a PD-L1-dependentin vitro bioassay method was set up to characterize GST-PD-1 bioactivity. The results suggested that GST-PD-1 could competently block the interaction between PD-Ll and PD-l and increase the production of IL-2 and IFN-γ of phytohemagglutinin-activated T cells.

  20. Mode of delivery and cord blood cytokines: a birth cohort study

    Directory of Open Access Journals (Sweden)

    DuBois Andrea M

    2006-09-01

    Full Text Available Abstract Background The mechanisms for the association between birth by cesarean section and atopy and asthma are largely unknown. Objective To examine whether cesarean section results in neonatal secretion of cytokines that are associated with increased risk of atopy and/or asthma in childhood. To examine whether the association between mode of delivery and neonatal immune responses is explained by exposure to the maternal gut flora (a marker of the vaginal flora. Methods CBMCs were isolated from 37 neonates at delivery, and secretion of IL-13, IFN-γ, and IL-10 (at baseline and after stimulation with antigens [dust mite and cat dander allergens, phytohemagglutinin, and lipopolysaccharide] was quantified by ELISA. Total and specific microbes were quantified in maternal stool. The relation between mode of delivery and cord blood cytokines was examined by linear regression. The relation between maternal stool microbes and cord blood cytokines was examined by Spearman's correlation coefficients. Results Cesarean section was associated with increased levels of IL-13 and IFN-γ. In multivariate analyses, cesarean section was associated with an increment of 79.4 pg/ml in secretion of IL-13 by CBMCs after stimulation with dust mite allergen (P Conclusion Cesarean section is associated with increased levels of IL-13 and IFN-γ, perhaps because of lack of labor and/or reduced exposure to specific microbes (e.g., gram-positive anaerobes at birth.

  1. PHA-induced inflammation is not energetically costly in the subterranean rodent Ctenomys talarum (tuco-tucos).

    Science.gov (United States)

    Merlo, Julieta L; Cutrera, Ana P; Luna, Facundo; Zenuto, Roxana R

    2014-09-01

    Immune activity has been proposed to be associated with substantial costs, due to trade-offs with other functions or activities that share common resources and contribute to an animal's fitness. However, direct estimates of the cost of mounting an immune response are few and have been performed mainly in birds. Thus, further work is needed to clarify the relative costs of different components of the immune system and the role of environmental and life-history traits in modulating the costs of resistance. Within the components of immunity, inflammation is considered to be associated with a larger energetic expenditure. Here, we evaluated the energetic cost of the inflammatory response to phytohemagglutinin (PHA) in a wild population of a subterranean rodent, Ctenomys talarum, and the trade-offs between immune activity and reproduction. C. talarum develops an inflammatory response to PHA, but contrary to our predictions, this response was not associated with an increase in oxygen consumption regardless of reproductive status or sex. Our study shows that an immune challenge may not always result in a detectable energetic cost. We discuss the possibility that other currencies could be underlying the cost, such as micro-or macronutrients requirements, autoimmunity or oxidative stress.

  2. Parasite infection negatively affects PHA-triggered inflammation in the subterranean rodent Ctenomys talarum.

    Science.gov (United States)

    Merlo, Julieta L; Cutrera, Ana P; Zenuto, Roxana R

    2016-02-01

    Magnitude and effectiveness of immune responses vary greatly between and within species. Among factors reported to determine this variation, parasitism is a critical one, although controversial effects of parasites over immunological indices have been reported. Information regarding immune strategies in species with different life histories is crucial to better understand the role of immune defenses in an ecological and evolutionary context. Here, we examine the influence of the parasite community on immune responsiveness of a solitary subterranean rodent, Ctenomys talarum. To do this, we assessed the impact of the natural parasite community and the experimental infection with Eimeria sp. on the phytohemagglutinin (PHA)-response, as well as other immune, condition, nutrition, and stress parameters. PHA-triggered inflammation was similarly impaired by Eimeria sp. infection alone or co-occurring with a number of gastrointestinal nematodes. None of the other physiological parameters studied were affected by parasitism. This indicates that parasitism is a general key factor modulating immune responsiveness of the host, and in particular for C. talarum, it could explain the great inter-individual variation previously observed in the PHA-response. Thus, our results highlight the importance of taking the parasite community into account in ecoimmunological studies, particularly when using immunological indices.

  3. Cytokine production and lymphocyte proliferation in patients with Nocardia brasiliensis actinomycetoma.

    Science.gov (United States)

    Méndez-Tovar, Luis J; Mondragón-González, Rafael; Vega-López, Francisco; Dockrell, Hazel M; Hay, Roderick; López-Martínez, Rubén; Manzano-Gayosso, Patricia; Hernández-Hernández, Francisca; Padilla-Desgarennes, Carmen; Bonifaz, Alexandro

    2004-11-01

    IFN-gamma, TNF-alpha, IL-4, IL-10 and IL-12 concentrations in the supernatant of peripheral blood mononuclear cell (PBMC) cultures and the in vitro proliferation of PBMC were studied in 25 patients with actinomycetoma caused by Nocardia brasiliensis and in 10 healthy controls from endemic zones. Cell cultures were stimulated by a N. brasiliensis crude cytoplasmic antigen (NB) and five semi-purified protein fractions (NB2, NB4, NB6, NB8, and NB10) separated by isoelectric. Phytohemagglutinin (PHA) and purified protein derivative (PPD) of Mycobacterium tuberculosis were used as control antigens. Skin tests were performed by injecting 0.1 ml of candidin and PPD intradermally (ID). Patients showed a poor response to tuberculin, while their response to candidin was more than two fold greater than that observed in the controls. Cell proliferation showed no statistically significant differences in either group. IFN-gamma production was higher in the healthy controls than in the patients, whereas TNF-alpha secretion was slightly higher in the patients' cultures. IL-4 was detected in the patients' cultures but not in the controls. IL-10 and IL-12 were present at low concentrations in both groups. These results suggest that patients with actinomycetoma show normal antigen recognition, but with low IFN-gamma production, and higher concentrations of IL-4, IL-10 and TNF-alpha in the patients' PBMC cultures, indicating that they probably have a Th2 type of immune response.

  4. Constitutive heterochromatin of chromosome 1 and Duffy blood group alleles in schizophrenia

    Energy Technology Data Exchange (ETDEWEB)

    Kosower, N.S.; Gerad, L.; Goldstein, M.; Parasol, N. [Tel-Aviv Univ. (Israel)] [and others

    1995-04-24

    Cytogenetic analysis was carried out in unrelated schizophrenic patients, unrelated controls and patients and family members in multiplex families. The size-distribution of chromosome 1 heterochromatic region (1qH, C-band variants) among 21 unrelated schizophrenic patients was different from that found in a group of 46 controls. The patient group had 1qH variants of smaller size than the control group (P < 0.01). Incubation of phytohemagglutinin-treated blood lymphocytes with 5-azacytidine (which causes decondensation and extension of the heterochromatin) led to a lesser degree of heterochromatin decondensation in a group of patients than in the controls (7 schizophrenic, 9 controls, P < 0.01). The distribution of phenotypes of Duffy blood group system (whose locus is linked to the 1qH region) among 28 schizophrenic patients was also different from that in the general population. Cosegregation of schizophrenia with a 1qH (C-band) variant and Duffy blood group allele was observed in one of six multiplex families. The overall results suggest that alterations within the Duffy/1qH region are involved in schizophrenia in some cases. This region contains the locus of D5 dopamine receptor pseudogene 2 (1q21.1), which is transcribed in normal lymphocytes. 33 refs., 1 fig., 2 tabs.

  5. Localization of the kappa opioid receptor gene to human chromosome band 8q11. 2

    Energy Technology Data Exchange (ETDEWEB)

    Yasuda, Kazuki; Takeda, Jun; Bell, G.I.; Espinosa, R.; Le Beau, M.M. (Univ. of Chicago, IL (United States))

    1994-02-01

    Using the cloned mouse kappa opioid receptor cDNA clone as a probe, screened a human genomic library and isolated a clone containing part of the human kappa opioid receptor gene (OPRK1), designated [lambda]hSR4-1. To determine the chromosomal localization of OPRK1, [lambda]hSR4-1 DNA was labeled with biotin by nick-translation in the presence of bio-11-dUTP and hybridized to human metaphase cells prepared from phytohemagglutinin-stimulated peripheral blood lymphocytes as described previously. Hybridization of the OPRK1-specific probe [lambda]hSR4-1 DNA to normal human metaphase chromosomes resulted in specific labeling only of chromosome 8. Specific labeling of 8q11 was observed on all 4 (6 cells), 3 (9 cells), 2 (9 cells), or 1 (1 cell) chromatid of the chromosome 8 homologs in 25 cells examined. Of 72 signals observed, 70 were located at 8q11. 1 signal was located at 7q11 and at 12p11. In most cells, the signal on 8q was located at 8q11.2. 7 refs., 1 fig.

  6. Chromosomal localization of the [delta] opioid receptor gene to human 1p34. 3-36. 1 and mouse 4D bands by in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Befort, K.; Kieffer, B. (Ecole Superieure de Biotechnologie de Strasbourg (France)); Mattei, M.G.; Roeckel, N. (Hopital d' Enfants de la Timone, Marseille (France))

    1994-03-01

    The aim of the present study is to determine the precise localization of this gene in the murine as well as human genome by in situ hybridization. Southern analysis, using the noncoding part of the cDNA as a probe (NotI-BamHI fragment, 1040 bp) under high-stringency conditions, shows the existence of a single-copy gene in the murine genome. In a similar analysis performed on human genomic DNA, the coding part of the cDNA (PstI-NotI fragment, 976 bp) clearly detected a single gene in the human genome (not shown). The authors therefore used these two probes for the chromosomal localization of the murine gene and its human counterpart, respectively. Chromosome spread preparations from concanavalin A (mouse) or phytohemagglutinin (human)-stimulated lymphocytes were hybridized with tritium-labeled cDNAs, exposed for 15 days, and developed, as described previously. For the murine assignment, a total of 100 metaphase cells were examined, and 149 silver grains were found. Forty-two grains were associated with chromosome 4, and 73.8% of them mapped to the D1-D3 region of the chromosome. In the 120 metaphase human cells analyzed, 197 silver grains were found, 25.8% of which were located on chromosome 1. The distribution of grains allowed mapping of the human [delta] opioid receptor gene to the p34.3-p36.1 region of the short arm with a maximum in the p35 band.

  7. Cytokines Expression and Nitric Oxide Production under Induced Infection to Typhimurium in Chicken Lines Divergently Selected for Cutaneous Hypersensitivity

    Directory of Open Access Journals (Sweden)

    Rani Singh

    2012-07-01

    Full Text Available In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h. The differential mRNA expression patterns of IFN-γ, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI as compared to the low cell-mediated line (LCMI upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-γ were observed in high response birds (HCMI; but IL-2 was down regulated in this line compared to the low response birds (LCMI. Significantly (p<0.05 higher expression of iNOS, IFN-γ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.

  8. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Energy Technology Data Exchange (ETDEWEB)

    Hill, F.S.; Cantu, A.O.; Lucas, J.N. (Lawrence Livermore National Lab., CA (United States)); Cox, A.B.; Salmon, Y.L. (Air Force Armstrong Lab., Brookes AFB, TX (United States))

    1994-10-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 [mu]g/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) that that produced by PHA (M1<0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture. (author).

  9. A galactose-inhibitable mitogen for human lymphocytes from the sponge axinella polypoides.

    Science.gov (United States)

    Phillips, S G; Bretting, H; Kabat, E A

    1976-10-01

    Of two galactose-binding hemagglutinins isolated from the sponge Axinella polypoides, axinella I was strongly mitogenic for human peripheral blood lymphocytes, and axinella II was not. Purified T cells responded strongly and B cells weakly to axinella I. Mitogenic response, as monitored by rate of 3H-thymidine incorporation on the third day of culture, was specifically inhibited by Dgalactose, Dfucose, raffinose, or 2-deoxy-D-galactose added within 5 hr of the mitogen. Mitogenic response was correlated with degree of lymphocyte agglutination. The effectiveness of a given sugar in inhibiting mitogenic response to axinella I paralleled its potency in inhibiting precipitation of lectin by blood group substances. If an inhibitory concentration of Dgalactose was add 24 to 40 hr after mitogenic activation, rate of 3H-thymadine uptake at 72 hr was two to twenty times above the rate induced in cultures to which no galactose was added. Dgalactose at a subinhibitory concentration (10mug/ml) enhanced 3H-thymidine incorportion incorporation induced by phytohemagglutinin or Con A, an effect reversible by Dgalactose. These findings suggest that axinella I has tow antagonistic effects on human lymphocytes: a) mitogenic activation and b) depressive activity resulting from depletion of essential galactose moieties.

  10. Comparative characteristics of human interleukin-2 preparations obtained from various sources

    Energy Technology Data Exchange (ETDEWEB)

    Iobadze, M.S.; Kulikov, V.V.; Kupriyanova, T.A.; Bykovskaya, S.N.; Bakhutashvili, V.I.

    1987-02-20

    Interleukin-2 (IL-2) was produced from donor peripheral blood lymphocytes and JURKAT FHCRC T lymphoma cells. Gel filtration, ion exchange chromatography on DEAE- and CM-Sephadex were used to purify the preparation. As a result of the purification, the specific activity of the preparation increased by a factor of 400. It was shown that optimum proliferation of T lymphocytes requires the successive action of phytohemagglutinin and IL-2, as well as the presence of serum in the medium. The properties and methods of production of a long-proliferating line of IL-2-dependent T cells B-5 are described. The proliferation of B-5 cells depends completely on the presence of IL-2 in the medium, although prolonged proliferation requires periodic stimulation by antigen (allogeneic lymphocytes). In the absence of IL-2 in the medium, B-5 cells die within 36 h. The prospects for the use of IL-2 preparations from human peripheral blood lymphocyte culture fluid for adoptive immunotherapy of tumors and the use of cells of the IL-2-dependent line B-5 in the testing of the activity of IL-2 preparations obtained from various sources are discussed.

  11. Protection Against Lung Cancer Patient Plasma-Induced Lymphocyte Suppression by Ganoderma Lucidum Polysaccharides

    Directory of Open Access Journals (Sweden)

    Li-Xin Sun

    2014-01-01

    Full Text Available Background/Aims: This study was conducted to determine the potential of Ganoderma lucidum polysaccharides (Gl-PS in protection against lung cancer patient plasma-induced suppression of lymphocytes. Lung cancer is a major cause of disease and loss of life in the United States and worldwide. Cancer cells release immunosuppressive mediators, such as PGE2, TGF-β, IL-10, and VEGF, to inhibit the immune response to escape from immune surveillance. Gl-PS has been shown to counteract this immune inhibition in an animal cell culture model, and thus to facilitate tumor control. The present study explored whether or not such an effect could also be demonstrated in human lung cancer patients. Methods: Immunofluorescence, flow cytometry, MTT, immunocytochemistry, and western blot analysis were used to assess lymphocyte activation with PHA. Results: The plasma of lung cancer patients suppressed proliferation, CD69 expression, and perforin and granzyme B production in lymphocytes upon activation by PHA, effects that were partially of fully reversed by Gl-PS. Conclusion: Lung cancer patient plasma-induced suppression of lymphocyte activation by phytohemagglutinin may be antagonized fully or partially by Gl-PS, an observation suggesting the potential of Gl-PS in cancer therapy.

  12. Distinct Overexpression of Fas Ligand on T Lymphocytes in Aplastic Anemia

    Institute of Scientific and Technical Information of China (English)

    Wenxin Li; Jinxiang Fu; Fengming Wang; Gehua Yu; Yong Wang; Xueguang Zhang

    2004-01-01

    Increased expression of Fas by hematopoietic progenitors in aplastic anemia (AA) suggests that Fas/Fas ligand (FasL) system plays a key role in the formation of severe pancytopenia. To further confirm the above hypothesis, T cells from 8 patients with AA were systematically studied for their FasL's distribution pattern,releasing manner and proapoptotic activity, compared with normal resting T cells and artificially activated Tcell blasts. The results demonstrated that AA T cells abnormally expressed low levels of membrane-bound FasL and contained high levels of intracellular FasL which could be triggered to release by high-dose phytohemagglutinin (PHA) pulse-stimulation. The supernatants from the PHA-stimulated AA T cells had apparent cytotoxicity against FasL-sensitive Jurkat cells, which could be significantly inhibited by monoclonal antibody against FasL in a dose-dependent manner, or nearly completely abrogated by ultracentrifugation. The above phenomena also appeared on artificially activated T cell blasts, but this was not the case on normal resting Tcells. These results indicate that AA T cell is a type of "preactivated" T lymphocyte, characterized by overexpression of FasL, especially intracellular FasL which can be stimulated to release in bioavtive exosomesbound form. Taken together, our data provide further and direct evidence for the hypothesis that T cells might mediate the destruction of hematopietic progenitor in AA through Fas/FasL system.

  13. Differential sensitivity of T lymphocytes and hematopoietic precursor cells to photochemotherapy with 8-methoxypsoralen and ultraviolet A light.

    Science.gov (United States)

    Mabed, Mohamed; Coffe, Christian; Racadot, Evelyne; Angonin, Regis; Pavey, Jean-Jaques; Tiberghien, Pierre; Herve, Patrick

    2006-01-01

    The combination of 8-methoxypsoralen (8-MOP) and long wave ultraviolet radiation (UV-A) has immunomodulatory effects and might abolish both graft-vs-host and host-vs-graft reactions after allogeneic hematopoietic stem cell transplantation. In the present study, we have confirmed the sensitivity of T lymphocytes to 8-MOP treatment plus UV-A exposure as evidenced by the abrogation of the alloreactivity in mixed lymphocyte cultures as well as the inhibition of the response to phytohemagglutinin A. However, the clonogenic capacity of the bone marrow hematopoietic progenitors was inhibited with UV-A doses lower than the doses needed to inhibit T-lymphocytes alloreactivity. Moreover, long-term bone marrow cultures showed that 8-MOP plus UV-A treatment had detrimental effects on the more immature bone marrow stem cells. These data were confirmed when murine bone marrow graft was treated with 8-MOP, exposed to UV-A, then transplanted into semiallogeneic recipient mice. The treated cells could not maintain their clonogenic capacity in vivo resulting in death of all animals. Taken together, these data show that ex vivo 8-MOP plus UV-A treatment of the marrow graft cannot be used to prevent post-bone marrow transplantation alloreactivity.

  14. CD8+ T cells of chronic HCV-infected patients express multiple negative immune checkpoints following stimulation with HCV peptides.

    Science.gov (United States)

    Barathan, Muttiah; Mohamed, Rosmawati; Vadivelu, Jamuna; Chang, Li Yen; Vignesh, Ramachandran; Krishnan, Jayalakshmi; Sigamani, Panneer; Saeidi, Alireza; Ram, M Ravishankar; Velu, Vijayakumar; Larsson, Marie; Shankar, Esaki M

    2017-03-01

    Hepatitis C virus (HCV)-specific CD4+ and CD8+ T cells are key to successful viral clearance in HCV disease. Accumulation of exhausted HCV-specific T cells during chronic infection results in considerable loss of protective functional immune responses. The role of T-cell exhaustion in chronic HCV disease remains poorly understood. Here, we studied the frequency of HCV peptide-stimulated T cells expressing negative immune checkpoints (PD-1, CTLA-4, TRAIL, TIM-3 and BTLA) by flow cytometry, and measured the levels of Th1/Th2/Th17 cytokines secreted by T cells by a commercial Multi-Analyte ELISArray™ following in vitro stimulation of T cells using HCV peptides and phytohemagglutinin (PHA). HCV peptide-stimulated CD4+ and CD8+ T cells of chronic HCV (CHC) patients showed significant increase of CTLA-4. Furthermore, HCV peptide-stimulated CD4+ T cells of CHC patients also displayed relatively higher levels of PD-1 and TRAIL, whereas TIM-3 was up-regulated on HCV peptide-stimulated CD8+ T cells. Whereas the levels of IL-10 and TGF-β1 were significantly increased, the levels of pro-inflammatory cytokines IL-2, TNF-α, IL-17A and IL-6 were markedly decreased in the T cell cultures of CHC patients. Chronic HCV infection results in functional exhaustion of CD4+ and CD8+ T cells likely contributing to viral persistence.

  15. A New Synthetic Compound, 2-OH, Enhances Interleukin-2 and Interferon-γ Gene Expression in Human Peripheral Blood Mononuclear Cells

    Directory of Open Access Journals (Sweden)

    Woan-Fang Tzeng

    2009-07-01

    Full Text Available A new synthetic compound, 6-hydroxy-2-tosylisoquinolin-1(2H-one (2-OH, was selected for immunopharmacological activity tests. The effects of 2-OH on human peripheral blood mononuclear cell (PBMC proliferation were determined by tritiated thymidine uptake. Compared to phytohemagglutinin (PHA; 5 μg/mL stimulation, 2-OH significantly enhanced PBMC proliferation in a dose-dependent manner. The 50% enhancement activity (EC50 for 2-OH was 4.4±0.1 μM. In addition, effects of 2-OH on interleukin-2 (IL-2 and interferon-γ (IFN-γ production in PBMC were determined by enzyme immunoassay. Results demonstrated that 2-OH stimulated IL-2 and IFN-γ production in PBMC. Data from reverse transcription-polymerase chain reaction (RT-PCR and real-time PCR indicated that IL-2 and IFN-γ mRNA expression in PBMC could be induced by 2-OH. Therefore, 2-OH enhanced IL-2 and IFN-γ production in PBMC by modulation their gene expression. We suggest that 2-OH may be an immunomodulatory agent.

  16. Central stimulation of hormone release and the proliferative response of lymphocytes in humans.

    Science.gov (United States)

    Juránková, E; Jezová, D; Vigas, M

    1995-01-01

    The central nervous system (CNS) may communicate with the immune system by direct innervation of lymphoid organs and/or by neurotransmitters and changes in neuroendocrine functioning and hormone release. The consequences of selective transient changes in circulating hormones on immune functioning in humans have not yet been studied. To address this problem, the authors evaluated the lymphoproliferative responses to optimal and suboptimal concentrations of phytohemagglutinin (PHA) and pokeweek mitogen (PWM) under selective enhancement of circulating growth hormone, prolactin, or norepinephrine. The authors failed to demonstrate any effect of elevated growth hormone levels after clonidine challenge on the lymphoproliferative response to mitogens. Similarly, the results did not show any effect of elevated prolactin concentrations induced by domperidone administration on the immune test. Exposure of volunteers to cold resulted in elevation of plasma norepinephrine levels without changes in growth hormone, epinephrine, or cortisol secretion. Cold exposure induced elevation of plasma norepinephrine and reduction of the lymphoproliferative response to the suboptimal dosage of PHA. The reduction was significant 180 and 240 min after exposure. These results are indicative of a relationship between norepinephrine and immunity.

  17. Analysis of cell cycle's correlation of γ-H2AX%γ-H2AX细胞周期相关性的分析

    Institute of Scientific and Technical Information of China (English)

    Yangping Yue; Jianping Gong; Zhenchuang Zhu; Dongdong Yu; Yu Deng; Dan Huang; Xiaolan Li; Wei Xiao; Deding Tao; Junbo Hu

    2008-01-01

    Objective: To analyze and discuss cell cycle's correlation of γ-H2AX, so as to accumulate the data for the further studies of γ-H2AX. Methods: MOLT-4 cells, and peripheral blood lymphocytes (PBLs), with or without 48 h stimulation of phytohemagglutinin (PHA), were irradiated by ultraviolet rays (UV rays). Fluorescence-labeled γ-H2AX antibody was used to detect γ-H2AX foci at the DNA double-strand breaks (DSBs) in chromatin, DNA damage was analyzed by flow cytometry, cell cycle and cell apoptosis were detected by sub-G1 peak method, the expression of y-H2AX was detected by Western blot. Results: With the progression of time, sub-G1 peak emerged apparently in the DNA histograms, and the cells of apoptosis increased gradually; with the progression of time, the increase of γ-H2AX emerged and firstly raised, then decreased; PBLs with 48 h stimulation of PHA entered apparently cell cycle, cells of S and G2/M phase emerged, and PBLs without stimulation of PHA did not enter cell cycle; Western blot showed the increase of the expression of γ-H2AX, and the increase also firstly raised, then decreased. Conclusion: γ-H2AX expressed in the cells of stationary phase and proliferative phase, and with the progression of time, the increase of γ-H2AX firstly raised, and then decreased.

  18. The Effect of Long-Term Exercise on the Production of Osteoclastogenic and Antiosteoclastogenic Cytokines by Peripheral Blood Mononuclear Cells and on Serum Markers of Bone Metabolism

    Directory of Open Access Journals (Sweden)

    J. Kelly Smith

    2016-01-01

    Full Text Available Although it is recognized that the mechanical stresses associated with physical activity augment bone mineral density and improve bone quality, our understanding of how exercise modulates bone homeostasis at the molecular level is lacking. In a before and after trial involving 43 healthy adults, we measured the effect of six months of supervised exercise training on the spontaneous and phytohemagglutinin-induced production of osteoclastogenic cytokines (interleukin-1α, tumor necrosis factor-α, antiosteoclastogenic cytokines (transforming growth factor-β1 and interleukins 4 and 10, pleiotropic cytokines with variable effects on osteoclastogenesis (interferon-γ, interleukin-6, and T cell growth and differentiation factors (interleukins 2 and 12 by peripheral blood mononuclear cells. We also measured lymphocyte phenotypes and serum markers of bone formation (osteocalcin, bone resorption (C-terminal telopeptides of Type I collagen, and bone homeostasis (25 (OH vitamin D, estradiol, testosterone, parathyroid hormone, and insulin-like growth factor 1. A combination of aerobic, resistance, and flexibility exercises done on average of 2.5 hours a week attenuated the production of osteoclastogenic cytokines and enhanced the production of antiosteoclastogenic cytokines. These changes were accompanied by a 16% reduction in collagen degradation products and a 9.8% increase in osteocalcin levels. We conclude that long-term moderate intensity exercise exerts a favorable effect on bone resorption by changing the balance between blood mononuclear cells producing osteoclastogenic cytokines and those producing antiosteoclastogenic cytokines. This trial is registered with Clinical Trials.gov Identifier: NCT02765945.

  19. Successful treatment for West syndrome with severe combined immunodeficiency.

    Science.gov (United States)

    Motobayashi, Mitsuo; Inaba, Yuji; Fukuyama, Tetsuhiro; Kurata, Takashi; Niimi, Taemi; Saito, Shoji; Shiba, Naoko; Nishimura, Takafumi; Shigemura, Tomonari; Nakazawa, Yozo; Kobayashi, Norimoto; Sakashita, Kazuo; Agematsu, Kazunaga; Ichikawa, Motoki; Koike, Kenichi

    2015-01-01

    Several immune mechanisms are suspected in the unknown etiology of West syndrome (WS). We report a male infant who suffered from WS and X-linked T-B+NK- severe combined immunodeficiency (X-SCID) with a missense mutation of the IL2RG gene (c.202G>A, p.Glu68Lys). He promptly began vitamin B6 and valproic acid treatment, but infantile spasms (IS) and hypsarrhythmia persisted. Administration of intravenous immunoglobulin and the change to topiramate (TPM) at 7 months of age resulted in the rapid resolution of IS. The CD4/8 ratio in his peripheral blood increased from 0.04-0.09 to 0.20-1.95 following unrelated cord blood transplantation (UCBT). In vitro lymphocyte proliferation in response to phytohemagglutinin or concanavalin A and the ability of B lymphocytes to produce antibodies improved as well. Electroencephalogram findings became normal 1 month after UCBT. Thus, we consider that T-cell dysfunction and/or impairments in T-B cell interactions due to X-SCID may have played important roles in the onset of WS. Immune-modulating therapies along with the administration of TPM effectively treated this severe epileptic syndrome in our patient.

  20. Immune response varies with rate of dispersal in invasive cane toads (Rhinella marina.

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    Gregory P Brown

    Full Text Available What level of immunocompetence should an animal maintain while undertaking long-distance dispersal? Immune function (surveillance and response might be down-regulated during prolonged physical exertion due to energy depletion, and/or to avoid autoimmune reactions arising from damaged tissue. On the other hand, heightened immune vigilance might be favored if the organism encounters novel pathogens as it enters novel environments. We assessed the links between immune defense and long-distance movement in a population of invasive cane toads (Rhinella marina in Australia. Toads were radio-tracked for seven days to measure their activity levels and were then captured and subjected to a suite of immune assays. Toads that moved further showed decreased bacteria-killing ability in their plasma and decreased phagocytic activity in their whole blood, but a heightened skin-swelling response to phytohemagglutinin. Baseline and post-stress corticosterone levels were unrelated to distance moved. Thus, long-distance movement in cane toads is associated with a dampened response in some systems and enhanced response in another. This pattern suggests that sustained activity is accompanied by trade-offs among immune components rather than an overall down or up-regulation. The finding that high mobility is accompanied by modification of the immune system has important implications for animal invasions.

  1. Sitagliptin treatment of patients with type 2 diabetes does not affect CD4+ T-cell activation.

    Science.gov (United States)

    White, Perrin C; Chamberlain-Shea, Heidi; de la Morena, Maria-Teresa

    2010-01-01

    Dipeptidyl peptidase IV (DPP4) inhibitors have recently become widely used for treating type 2 diabetes, but in meta-analyses are associated with a mildly increased risk of all-cause infections. CD26 is a cell-surface form of DPP4 which can costimulate T-cell proliferation, raising the possibility that DPP4 inhibitors might adversely affect immune function. To address this issue in an observational study, two groups of 20 subjects each were recruited from a private endocrinology practice; one group consisted of type 2 diabetes patients treated for at least 6 months with the DPP4 inhibitor, sitagliptin, whereas patients in the other group had never been treated with this agent. The groups were similar with regard to sex and racial composition, body mass index, hemoglobin A(1c), and use of other medications for diabetes, but the sitagliptin group was slightly older. A blood sample from each patient was analyzed for CD4+ T-cell activation in response to phytohemagglutinin using adenosine triphosphate (ATP)-stimulated bioluminescence. There was not a significant difference in T-cell activation between the treatment groups (median, 419 and 481 ng/ml ATP in the groups that were and were not treated with sitagliptin, respectively). Thus the observed increased rate of infection in diabetic patients treated with sitagliptin cannot be explained by a major effect on T-cell activation. Randomized studies, preferably using several assays of immune function, should be performed to confirm and extend these findings.

  2. Food Restriction Affects Inflammatory Response and Nutritional State in Tuco-tucos (Ctenomys talarum).

    Science.gov (United States)

    Merlo, Julieta Leticia; Cutrera, Ana Paula; Zenuto, Roxana Rita

    2016-12-01

    Insufficient or unbalanced food intake typically has a negative impact on immune responses. The understanding of this effect is, however, hampered by the effect that food has on general condition, which, in turn, affects immunity, and the interaction among general condition, immunocompetence, and concurrent infections. The goal of this study was to determine the effects of food restriction and methionine supplementation on immunity in tuco-tucos (Ctenomys talarum). Effects of diet manipulations on nutritional state, inflammatory response to phytohemagglutinin (PHA), and other immune parameters (bacterial killing capacity, natural antibodies, and leukocyte profile) were evaluated. Health and stress parameters and endoparasite loads were assessed to understand more deeply potential effects of treatments on immune status. Individuals under food restriction presented an altered nutritional state as well as increased stress levels (higher N: L ratios) compared with individuals fed ad libitum, and a marked reduction in the inflammatory response to PHA. Supplementation with methionine did not affect any of the parameters analyzed. Endoparasite loads were not affected by treatments. Our results support the idea that food insufficiency can modulate the individual's immune responsiveness through the lack of adequate essential nutrients, metabolic fuel and energetic reserves, or by a detrimental effect of the stress caused by nutrient limitation. We show that the response to PHA previously reported as nonenergetically costly for C. talarum, implies a nutritional cost; an opposite pattern to that previously found for the adaptive antibody response to sheep red blood cells in the same species.

  3. Antigen-specific secretion of IFNγ and CXCL10 in whole blood assay detects Mycobacterium leprae infection but does not discriminate asymptomatic infection from symptomatic leprosy.

    Science.gov (United States)

    Hungria, Emerith Mayra; Freitas, Aline Araújo; Pontes, Maria Araci Andrade; Gonçalves, Heitor Sá; Sousa, Ana Lúcia Osório Maroccolo; Costa, Maurício Barcelos; Castilho, Mirian Lane Oliveira Rodrigues; Duthie, Malcolm S; Stefani, Mariane Martins Araújo

    2017-04-01

    To advance toward a whole blood assay (WBA)-based test capable of facilitating the diagnosis of paucibacillary (PB) leprosy, we evaluated a prototype in-tube WBA using combinations of Mycobacterium leprae antigens. Blood was collected from newly diagnosed untreated PB (n=38), multibacillary (MB) (n=30), healthy household contacts (HHC) of MB (n=27), and endemic controls (n=61) residing in Goiânia and Fortaleza, Brazil. Blood was incubated with M. leprae cell sonicate, recombinant proteins (46f+LID-1; ML0276+LID-1), or controls (phosphate-buffered saline, phytohemagglutinin, M. tuberculosis purified protein derivative). Antigen-specific IFNγ production was observed in 71-84% and 55% of PB and HHC, respectively. Antigen-specific CXCL10 levels were similarly assessed to determine if, unlike IFNγ, CXCL10 could differentiate PB from HHC with repeated exposure/asymptomatic M. leprae infection. The CXCL10 levels induced in response to M. leprae antigens could not, however, differentiate PB from HHC. Despite these limitations, the WBAs reported here still represent important tools for assessing M. leprae infection rates and evaluating the impact of control measures.

  4. Immunotherapy of gastrointestinal cancer patients with levamisole.

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    Miwa,Hiroaki

    1979-02-01

    Full Text Available Levamisole was administered to 177 patients with gastrointestinal cancer (88 curative resection, 58 noncurative resection and 31 without resection. It was administered at a daily dose of 150 mg for three consecutive days every other week. The administration was started, as a rule, 3 days before operation. This medication was repeated as frequently as possible at least for one month. The cellular immunity and 18-month survival rate of treated and control groups were compared. Levamisole effectively improved peripheral lymphocyte blastformation against phytohemagglutinin and increased the numbers of peripheral blood lymphocytes. Levamisole caused extremely high blastformation rates, in general, enhanced PPD reactions in non-curative resection cases 7 months after operation and showed no influence upon the number of peripheral blood lymphocyte. The effect of levamisole on the 6-month survival rate was most marked in patients without resection. Increased 12-month survival rate was marked in non-curative resection cases and, to a lesser extent, curative resection cases. Patients without resection had a slightly improved 12-month survival rate. Levamisole improved the 18-month survival rate in resectable cases; however, there were no significant differences in 18-month survival between levamisole and control groups of patients not undergoing resection. The results suggest that levamisole is effective in the patients whose tumor cells have been decreased by any method.

  5. Human immunodeficiency virus type 1 RNA detection in peripheral blood mononuclear cells by polymerase chain reaction: enhanced sensitivity after mitogenic stimulation.

    Science.gov (United States)

    Tetali, S K; Oyaizu, N; Paul, M; Pahwa, S

    1993-01-01

    The aim of this study was to investigate whether stimulus-induced up-regulation of human immunodeficiency virus type 1 (HIV-1) expression in peripheral blood mononuclear cells (PBMC) could enhance the diagnostic sensitivity of the polymerase chain reaction (PCR). PBMC derived from 11 HIV-1-infected asymptomatic adults were cultured with a stimulus of phytohemagglutinin (PHA) plus phorbol 12-myristate 13-acetate (PMA) for 36 h prior to lysing the cells for PCR. In all 11 patients studied, the intensity of PCR-assisted HIV RNA amplification (RNA-PCR) performed on stimulated cells was significantly (p < 0.001) higher than that obtained on unstimulated cells. A comparison of conventional PCR-assisted DNA amplification (DNA-PCR) with that of RNA-PCR was made on seven patients. The sensitivity of DNA-PCR was also increased by prior stimulation of cells, although not to the same extent as was observed for RNA-PCR. The results of our study indicate that the sensitivity of PCR can be significantly enhanced by prior activation of cells with PHA and PMA.

  6. In Vitro Evaluation of Colloidal Silver on Immune Function: Antilymphoproliferative Activity

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    M. A. Franco-Molina

    2016-01-01

    Full Text Available Colloidal silver (AgC is currently used by humans and it can be internalized through inhalation, injection, ingestion, and dermal contact. However, there is limited information about immunological activity; more investigations using colloidal silver are needed. In the present study, the effects of AgC (17.5 ng/mL on immunological parameters (proliferation and immunophenotyping using human peripheral blood mononuclear cells (PBMC and macrophages (phagocytosis and cytotoxicity on leukemia and lymphoma cancer cell lines (1.75 to 17.5 ng/mL were investigated. AgC was observed to significantly (p<0.05 decrease interleukin-2 (IL-2 production and proliferation induced by phytohemagglutinin or concanavalin A in PBMC without affecting its cell viability but with cytotoxic effect on cancer cells. IL-2, IL-4, IL-6, IL-10, INF-γ, and IL-17A cytokines production and CD3+, CD3−CD19+, CD3+CD4+, CD3+CD8+, and CD16+CD56+ PBMC phenotypes were not affected by AgC. The present study demonstrates that colloidal silver is harmless and nontoxic to the immune system cells and its ability to interfere with the immune response by decreasing cell proliferation when stimulated with mitogens demonstrated the antilymphoproliferative potential of AgC.

  7. Intrahepatic and peripheral T-cell responses in genotype 1b hepatitis C virus-infected patients with persistently normal and elevated aminotransferase levels

    Institute of Scientific and Technical Information of China (English)

    Filiz Akyüz; Nuray Polat; Sabahattin Kaymakoglu; Nevzat Aksoy; Kadir Demir; Fatih Be(s)i(s)ik; Selim Badur; Yilmaz (C)akaloglu; Atilla (O)kten

    2005-01-01

    AIM: To evaluate whether the cytokine responses in liver and serum differ in chronic hepatitis C patients with normal and high alanine aminotransferase (ALT) levels.METHODS: Thirty-three (16 with normal ALT level as group 1 and 17 with elevated ALT level as group 2) patients infected with genotype 1b hepatitis C virus (HCV) were examined. Liver infiltrating lymphomononuclear cells (LILMCs) were isolated from liver biopsy by collagenase type 1 and stimulated with phytohemagglutinin and interleukin 2 (IL-2). IL-10, IL-12,interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) were determined in serum and LILMCs by ELISA.RESULTS: Serum cytokine levels were similar in both groups (P>0.05). Stimulated IFN-γ and TNF-α levels in LILMCs were increased in both groups. IL-12 and IL-10levels stimulated with IL-2 were higher in group 1 than in group 2 (P = 0.023). Histological activity index (HAI)and stage had a negative correlation with TNF-α and IFN-γ levels in group 2.CONCLUSION: Increased T-helper type 2 (Th2)cytokine response may regress inflammatory and biochemical activity. Progression of histological abnormalities in persons with elevated ALT probably depends on insufficient Th2 cytokine response, which does not balance Th1 cytokine response.

  8. The Study on Immuno-modulation of Triptolide in vitro

    Institute of Scientific and Technical Information of China (English)

    翁小满; S.Atkinson; P.Gorak-Stolinska; H.M.Dockrell

    2004-01-01

    The purpose of this investigation is to demonstrate whether triptolide can inhibit TNF-α selectively and to study the mechanism. The ability of triptolide to inhibit the production of TNF-α and IFN-γ stimulated by lipopolysaccharide (LPS) and phytohemagglutinin (PHA) on peripheral blood mononuclear cell (PBMC) from healthy donors was measured by ELISA assays. The immunological mechanism of action of triptolide on TNF-α was investigated by pre-treatment with triptolide of PBMC and monecytes followed by analysis with Flow Cytometry (FCM). The inhibition of TNF-α and IFN-γ by triptol-ide occurred in a dose dependent manner and the IC50 was equal to 5-10 ng/ml for TNF-a and 0.1-1 ng/ml for IFN-γ. The concentrations of TNF-α measured after the different pre-treatments with triptolide on PBMC and monecytes are consistentwith its effects on a population of CD14+/TNF-α monecytes shown on FCM. The two methods of pre-treatments with triptolide may suggest different clinical significances, Immunophenotyping analysis with FCM revealed that triptolide may compete with LPS for binding to the CD14 receptor. The study provides basic evidence that triptolide may be used as an anti-inflam-matory reagent for treatment of leprosy reactions.

  9. Inter- and intra-individual variation in tests of cell-mediated immunity in young and old women.

    Science.gov (United States)

    Molls, Roshni R; Ahluwalia, Namanjeet; Fick, Tara; Mastro, Andrea M; Wagstaff, David; Handte, Gordon; Ball, Rick

    2003-05-01

    Exploring means to maintain or improve immunity in older persons has been receiving attention. To establish relationships between immune function and variables of interest, it is important to determine these variables accurately and precisely. Precision relates to the degree of variation in the laboratory test. The nature and magnitude of variability in tests of immune function has not been described extensively. We examined inter- and intra-individual variation in tests of cell-mediated immunity (CMI) in generally healthy and well-nourished young (20-40 years; n=15) and old (60-80 years; n=15) women. Subjects provided blood samples on 2 days within a week to determine leukocyte subsets, T-cell proliferation response to phytohemagglutinin A and concanavalin A, and interleukin (IL)-1beta, IL-2 and IL-6 production by stimulated mononuclear cells. Intra-individual variation was partitioned into day-to-day biological and analytical variation. Inter-individual variation was greater than intra-individual variability for most tests of CMI for both age groups. Furthermore, all CMI tests exhibited large day-to-day intra-individual variation (CV approximately 15% or greater) which was primarily due to biological rather than analytical sources, for both age groups. In conclusion, both age groups showed large between-person and considerable within-person variation in CMI tests. Therefore, assessment of CMI based on a single blood draw may not provide a reliable estimate of immune function.

  10. Identification of a novel splice variant of human PD-L1 Mrna encoding an isoform-lacking Igv-like domain

    Institute of Scientific and Technical Information of China (English)

    Xian-hui HE; Li-hui XU; Yi LIU

    2005-01-01

    Aim: To investigate the expression and regulation of PD-1 ligand 1 (PD-L1) in peripheral blood mononuclear cells (PBMC). Methods: The cDNA encoding human PD-L1 precursor was cloned from the total RNA extracted from the resting and phorbol dibutyrate plus ionomycin- or phytohemagglutinin-activated PBMC, by reverse transcription polymerase chain reaction (RT-PCR), and independent clones were sequenced and analyzed. The expression and subcellular localization were examined in transiently transfected cells. The PD-L1 gene expression in different PBMC was also analyzed by RT-PCR. Results: A novel human PD-L1 splice variant was identified from the activated PBMC. It was generated by splicing out exon 2 encoding an immunoglobulin variable domain (Igv)-like domain but retaining all other exons without a frame-shift. Consequently, the putative translated protein contained all other domains including the transmembrane region except for the Igv-like domain. Furthermore, the conventional isoform was expressed on the plasma surface whereas the novel isoform showed a pattern of intmcellular membrane distribution in transiently transfected K562 cells. In addition, the expression pattern of the PD-L1 splice variant was variable in different individuals and in different cellular status. Conclusion: PD-L1 expression may be regulated at the posttranscriptional level through alternative splicing, and modulation of the PD-L1 isoform expression may influence the outcome of specific immune responses in the peripheral tissues.

  11. Vitamin B12 and Folic Acid Imbalance Modifies NK Cytotoxicity, Lymphocytes B and Lymphoprolipheration in Aged Rats

    Directory of Open Access Journals (Sweden)

    Teresa Partearroyo

    2013-11-01

    Full Text Available Different vitamin B12 and folic acid concentrations could exacerbate the immune response. The aim was to evaluate different dietary folic acid and vitamin B12 levels on the immune response in aged rats. Male Sprague Dawley aged rats were assigned to three folic acid groups (deficient, control, supplemented each in absence of vitamin B12 for 30 days. Several parameters of innate and acquired immune responses were measured. Serum and hepatic folate levels increased according to folic acid dietary level, while vitamin B12 levels decreased. There was a significant decrease in natural killer cell-mediated cytotoxicity in the spleen for the vitamin B12 deficient diet and folic acid control diet groups. Significant changes in CD45 lymphocyte subsets were also observed according to dietary imbalance. Lymphoproliferative response to concanavalin A and phytohemagglutinin did not differ significantly between groups. The spleen response to lipopolysaccharide increased significantly, but was unmodified for the other organs. An imbalance between dietary vitamin B12 and folic acid concentrations alters some immunological parameters in aged rats. Therefore, the ratio between folate and vitamin B12 could be as important as their absolute dietary concentrations.

  12. The resistance of activated T-cells from SLE patients to apoptosis induced by human thymic stromal cells.

    Science.gov (United States)

    Budagyan, V M; Bulanova, E G; Sharova, N I; Nikonova, M F; Stanislav, M L; Yarylin, A A

    1998-01-01

    In this paper we show the differential sensitivity of phytohemagglutinine (PHA) activated T-cells from healthy donors or patients with systemic lupus erythematosus (SLE) to apoptosis induced by human thymic stromal cell line of epithelial origin. T-cells from SLE patients were mainly resistant to the apoptotic action of the stromal cells, while normal T-lymphocytes readily died via apoptosis. Gel electrophoresis revealed a DNA fragmentation pattern characteristic of apoptosis after 18 h of coculture. The simultaneous measurement of [3H]-thymidine uptake showed that the proliferative response of T-cells from SLE patients was significantly decreased compared to their normal counterparts. Such difference may account for the distinct result of interactions between the stromal and lymphoid cells, leading to the subsequent survival of T-lymphocytes from SLE patients. Nevertheless pretreatment of normal activated T-lymphocytes with anti-Fas mAbs, which have the capacity to substantially inhibit signaling through this receptor resulted in abolition of this form of programmed cell death. Thus, the precise role of Fas receptor and its ligand in this in vitro test system needs further investigation.

  13. Electrostimulation of rat callus cells and human lymphocytes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Aro, H.; Eerola, E.; Aho, A.J.; Penttinen, R.

    1984-01-01

    Asymmetrical pulsing low voltage current was supplied via electrodes to cultured rat fracture callus cells and human peripheral blood lymphocytes. The (/sup 3/H)thymidine incorporation of the callus cells and 5-(/sup 125/I)iodo-2'-deoxyuridine incorporation of the lymphocytes were determined. The growth pattern of callus cells (estimated by cellular density) did not respond to electrical stimulation. However, the uptake of (/sup 3/H)thymidine was increased at the early phase of cell proliferation and inhibited at later phases of proliferation. The (/sup 3/H)thymidine uptake of confluent callus cell cultures did not respond to electrical stimulation. Lymphocytes reacted in a similar way; stimulated cells took up more DNA precursor than control cells at the early phase of stimulation. During cell division, induced by the mitogens phytohemagglutinin and Concanavalin-A, the uptake of DNA precursor by stimulated cells was constantly inhibited. The results suggest that electrical stimuli affect the uptake mechanisms of cell membranes. The duality of the effect seems to be dependent on the cell cycle.

  14. Evaluation of Some Sugarcane (Saccharum officinarum L. Extracts for Immunostimulatory and Growth Promoting Effects in Industrial Broiler Chickens

    Directory of Open Access Journals (Sweden)

    Mian Muhammad Awais and Masood Akhtar*

    2012-06-01

    Full Text Available Present paper describes the immunostimulatory and growth promoting effects of some sugar cane extracts (SCEs in broiler chickens. Aqueous extract (AE from sugar cane (Saccharum officinarum juice and ethanolic extract (EE from bagasse were used to demonstrate their effects on lymphoproliferative responses to Phytohemagglutinin-P (PHA-P and Concanavalin-A (Con-A; antibody response to sheep red blood cells (SRBCs; growth rate and feed conversion ratio (FCR in experimental chickens as compared to control. Results showed significantly higher (P<0.05 in vitro and in vivo lymphoproliferative responses to Con-A and PHA-P, respectively in chickens administered with SCEs as compared to those in control group. Further, significantly higher (P<0.05 lymphoproliferative responses were detected in chickens administered with EE as compared to chickens administered with AE. Anti-SRBC total Igs, IgG and IgM titers were significantly higher (P<0.05 in chickens of experimental groups administered with SCEs as compared to those of control group; whereas titers were comparable among the experimental groups. The organ-body weight ratios of lymphoid organs were statistically similar in experimental and control groups. Both the experimental groups administered with SCEs showed better FCR and significantly higher (P<0.05 weight gains as compared to control. In conclusion, oral administration of SCEs showed immunostimulatory effects in broiler chickens and resulted in improved feed utilization and decreased amount of food needed for unit gain in body weight.

  15. Leptin enhances the release of cytokines by peripheral blood mononuclear cells from acute multiple sclerosis patients

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective To explore the effect of leptin on cytokine production by PBMCs obtained from MS patients either in acute (relapse) or in stable (nonrelapse) phase of disease. Methods PBMCs were collected from 25 untreated acute MS patients, 11 stable MS patients and 20 healthy controls. PBMCs were cultured either with RPMI-1640 alone or with leptin (1.25 nmol/ml), phytohemagglutinin (PHA) ( 100 μg/ml), and leptin + PHA. 72 h later the supernate of the culture medium were collected and stored at -70℃. The pro-inflammatory cytokine (IFN-γ) concentration were determined using an enzyme-linked immunosorbent assay ( ELISA), and the anti-inflammatory cytokine (IL-4) concentration were investigated by radioimmunity methods. Results Our data showed that leptin induced IFN-γproduction by PBMCs of patients in an acute phase of disease but not in a stable phase or in healthy controls. Moreover, we found that PHA induced IL-4 production by PBMCs of patients in an acute phase of disease, but leptin inhibited this ability of PHA. Conclusion Leptin can affect on pro- and anti-inflammatory cytokine production by PBMCs collected from MS patients, may be this connected with leptin increase the susceptiveness of MS.

  16. Cytokine production in patients with papillary thyroid cancer and associated autoimmune Hashimoto thyroiditis.

    Science.gov (United States)

    Zivancevic-Simonovic, Snezana; Mihaljevic, Olgica; Majstorovic, Ivana; Popovic, Suzana; Markovic, Slavica; Milosevic-Djordjevic, Olivera; Jovanovic, Zorica; Mijatovic-Teodorovic, Ljiljana; Mihajlovic, Dusan; Colic, Miodrag

    2015-08-01

    Hashimoto thyroiditis (HT) is the most frequent thyroid autoimmune disease, while papillary thyroid cancer (PTC) is one of the most common endocrine malignancies. A few patients with HT also develop PTC. The aim of this study was to analyze cytokine profiles in patients with PTC accompanied with autoimmune HT in comparison with those in patients with PTC alone or HT alone and healthy subjects. Cytokine levels were determined in supernatants obtained from phytohemagglutinin (PHA)-stimulated whole blood cultures in vitro. The concentrations of selected cytokines: Th1-interferon gamma (IFN-γ); Th2-interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 10 (IL-10) and interleukin 13 (IL-13); Th9-interleukin 9 (IL-9); and Th17-interleukin 17 (IL-17A) were measured using multiplex cytokine detection systems for human Th1/Th2/Th9/Th17/Th22. We found that PTC patients with HT produced significantly higher concentrations of IL-4, IL-6, IL-9, IL-13 and IFN-γ than PTC patients without HT. In conclusion, autoimmune HT affects the cytokine profile of patients with PTC by stimulating secretion of Th1/Th2/Th9 types of cytokines. Th1/Th2 cytokine ratios in PTC patients with associated autoimmune HT indicate a marked shift toward Th2 immunity.

  17. The effect of cocoa procyanidins on the transcription and secretion of interleukin 1 beta in peripheral blood mononuclear cells.

    Science.gov (United States)

    Mao, T K; Powell, J; Van de Water, J; Keen, C L; Schmitz, H H; Hammerstone, J F; Gershwin, M E

    2000-03-03

    Recent data has demonstrated that cacao liquor polyphenols (procyanidins) have antioxidant activity, inhibit mRNA expression of interleukin-2 and are potent inhibitors of acute inflammation. Given the widespread ingestion of cocoa in many cultures, we investigated whether cocoa, in its isolated procyanidin fractions (monomer through decamer), would modulate synthesis of the pro-inflammatory cytokine, interleukin-1 beta. Both resting and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) were investigated at the levels of transcription and protein secretion. Individual cocoa fractions were shown to augment constitutive IL-1 beta gene expression, although values varied between subjects. Interestingly, the smaller fractions of cocoa (monomer-tetramer) consistently reduced IL-1 beta expression of PHA-stimulated cells by 1-15%, while the larger oligomers (pentamer-decamer) increased expression by 4-52%. These data, observed at the transcription level, were reflected in protein levels in PHA-induced PBMC. The presence or absence of PHA did not alter the effects of the cocoa procyanidins with the exception of the pentamer. This study offers additional data for the consideration of the health-benefits of dietary polyphenols from a wide variety of foods, including those benefits associated specifically with cocoa and chocolate consumption.

  18. Optimized immobilization of lectins using self-assembled monolayers on polysilicon encoded materials for cell tagging.

    Science.gov (United States)

    Penon, Oriol; Siapkas, Dimitrios; Novo, Sergi; Durán, Sara; Oncins, Gerard; Errachid, Abdelhamid; Barrios, Lleonard; Nogués, Carme; Duch, Marta; Plaza, José Antonio; Pérez-García, Lluïsa

    2014-04-01

    Self-assembled monolayers (SAMs) have been used for the preparation of functional microtools consisting of encoded polysilicon barcodes biofunctionalized with proteins of the lectin family. These hybrid microtools exploit the lectins ability for recognizing specific carbohydrates of the cell membrane to give an efficient system for cell tagging. This work describes how the control of the methodology for SAM formation on polysilicon surfaces followed by lectin immobilization has a crucial influence on the microtool biofunction. Several parameters (silanization time, silane molar concentration, type of solvent or deposition methodology) have been studied to establish optimal function. Furthermore, silanes incorporating different terminal groups, such as aldehyde, activated ester or epoxide groups were tested in order to analyze their chemical coupling with the biomolecules, as well as their influence on the biofunctionality of the immobilized protein. Two different lectins - wheat germ agglutinin (WGA) and phytohemagglutinin (PHA-L) - were immobilized, because they have different and specific cell recognition behaviour and exhibit different cell toxicity. In this way we can assess the effect of intrinsic bulk toxicity with that of the cell compatibility once immobilized as well as the importance of cell affinity. A variety of nanometrical techniques were used to characterize the active surfaces, and lectin immobilization was quantified using ultraviolet-visible absorption spectroscopy (UV-vis) and optical waveguide light mode spectroscopy (OWLS). Once the best protocol was found, WGA and PHA were immobilized on polysilicon coded barcodes, and these microtools showed excellent cell tagging on living mouse embryos when WGA was used.

  19. "Unconventional" Neutralizing Activity of Antibodies Against HIV

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Neutralizing antibodies are recognized to be one of the essential elements of the adaptive immune response that must be induced by an effective vaccine against HIV. However, only a limited number of antibodies have been identified to neutralize a broad range of primary isolates of HIV-1 and attempts to induce such antibodies by immunization were unsuccessful. The difficulties to generate such antibodies are mainly due to intrinsic properties of HIV-1 envelope spikes, such as high sequence diversity, heavy glycosylation, and inducible and transient nature of certain epitopes. In vitro neutralizing antibodies are identified using "conventional" neutralization assay which uses phytohemagglutinin (PHA)-stimulated human PBMCs as target cells. Thus, in essence the assay evaluates HIV-1 replication in CD4+ T cells. Recently, several laboratories including us demonstrated that some monoclonal antibodies and HIV-1-specific polyclonal IgG purified from patient sera, although they do not have neutralizing activity when tested by the "conventional" neutralization assay, do exhibit potent and broad neutralizing activity in "unconventional" ways. The neutralizing activity of these antibodies and IgG fractions is acquired through post-translational modifications, through opsonization of virus particles into macrophages and immature dendritic cells (iDCs), or through expression of antibodies on the surface of HIV-1-susceptible cells. This review will focus on recent findings of this area and point out their potential applications in the development of preventive strategies against HIV.

  20. Recovery of immune competence following sublethal X irradiation of young and old mice: a model for studying age-related loss of immunologic homeostasis

    Energy Technology Data Exchange (ETDEWEB)

    Peterson, W.J. (VA Wadsworth Medical Center, Los Angeles, CA); Perkins, E.H.; Makinodan, T.

    1982-01-01

    Age-related alteration in lymphohematopoietic homeostasis was assessed kinetically by determining immunologic and stem-cell regenerating capacities of young (5-7 months), middle-aged (13 months), and old (23-24 months) C3H and C57BL/6 mice following their exposure to 500 R. Immunologic activities were based on the ability of spleen cells to respond to sheep erythrocytes, phytohemagglutinin, and bacterial lipopolysaccharide. Stem-cell activity was based on the ability of splenic and bone marrow cells to form colonies in vivo. Reflective of age-related homeostatic imbalance was alteration in the (a) time of recovery, (b) rate of regeneration, and (c) capacity of the regenerating system to overshoot the preirradition steady-state level. Most of the immunologic parameters showed a delay in the time of recovery in old mice. In contrast, the time of recovery of stem cells in old mice was equal to or faster than that in young mice. Furthermore, the magnitude of regeneration of stem cells was greater in old than young mice. These results suggest that recovery of immunologic activities in old mice is delayed partly because of the inability of their stem cells to rapidly generate immunocompetent progenies.

  1. Chromosome aberrations in human lymphocytes induced by 250 MeV protons: effects of dose, dose rate and shielding

    Science.gov (United States)

    George, K.; Willingham, V.; Wu, H.; Gridley, D.; Nelson, G.; Cucinotta, F. A.

    2002-01-01

    Although the space radiation environment consists predominantly of energetic protons, astronauts inside a spacecraft are chronically exposed to both primary particles as well as secondary particles that are generated when the primary particles penetrate the spacecraft shielding. Secondary neutrons and secondary charged particles can have an LET value that is greater than the primary protons and, therefore, produce a higher relative biological effectiveness (RBE). Using the accelerator facility at Loma Linda University, we exposed human lymphocytes in vitro to 250 MeV protons with doses ranging from 0 to 60 cGy at three different dose rates: a low dose rate of 7.5 cGy/h, an intermediate dose rate of 30 cGy/h and a high dose rate of 70 cGy/min. The effect of 15 g/cm2 aluminum shielding on the induction of chromosome aberrations was investigated for each dose rate. After exposure, lymphocytes were incubated in growth medium containing phytohemagglutinin (PHA) and chromosome spreads were collected using a chemical-induced premature chromosome condensation (PCC) technique. Aberrations were analyzed using the fluorescence in situ hybridization (FISH) technique with three different colored chromosome-painting probes. The frequency of reciprocal and complex-type chromosome exchanges were compared in shielded and unshielded samples. c2002 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

  2. Ultraviolet irradiation of platelet concentrates: Feasibility in transfusion practice

    Energy Technology Data Exchange (ETDEWEB)

    Andreu, G.; Boccaccio, C.; Lecrubier, C.; Fretault, J.; Coursaget, J.; LeGuen, J.P.; Oleggini, M.; Fournel, J.J.; Samama, M. (Secteur d' Hemobiologie Transfusion, Paris (France))

    1990-06-01

    Ultraviolet (UV)-B irradiation abolishes lymphocyte functions (the ability to respond and to stimulate) in mixed lymphocyte culture (MLC). This effect may have practical application in the prevention or reduction of transfusion-induced alloimmunization against HLA class I antigens. To study this, platelet concentrates (PCs) were obtained with a cell separator, suspended in autologous plasma in a final volume of 400 mL, and transferred into a large (22 X 30 cm) cell culture bag. This plastic showed a good transmittance of UV-B rays at 310 nm (54%). PCs were placed between two quartz plates (surface of irradiation = 25 X 37 cm), and the two sides were irradiated simultaneously. Energy delivered to the surface of the plastic bag was automatically monitored. The ability to respond (in MLC and to phytohemagglutinin) and to stimulate allogeneic lymphocytes was completely abolished with energy of 0.75 J per cm2 (irradiation time less than 3 min). The temperature increase during irradiation was negligible. Platelet aggregation (collagen, adrenalin, ADP, arachidonic acid, ristocetin) was not impaired if UV-B energy was below 3 J per cm2. Recovery and survival of autologous 111In-labeled platelets were studied in four volunteers; no differences were found between UV-B-treated (1.5 J/cm2) platelets and untreated platelets. These results show that a large-scale clinical trial using UV-B-irradiated PCs to prevent HLA alloimmunization is feasible.

  3. Evaluation of the immunotoxicity of low-level PCB (polychlorinated biphenyl) exposure in the rat

    Energy Technology Data Exchange (ETDEWEB)

    Smialowicz, R.J.; Andrews, J.E.; Riddle, M.M.; Rogers, R.R.; Luebke, R.W.

    1989-01-01

    Weanling male Fischer 344 rats were exposed daily by gastric intubation for up to 15 weeks to the polychlorinated biphenyl (PCB) Aroclor 1254 at 0.1, 1, 10, or 25 mg/kg body weight. At 5, 10 and 15 weeks groups of rats were killed and immune functions were evaluated. The immune parameters examined included the following: body and lymphoid organ weights, mitogen stimulated lymphoproliferative (LP) responses, natural killer (NK) cell activity, mixed lymphocyte reaction (MLR), and cytotoxic T lymphocyte (CTL) response. After 10 and 15 weeks of dosing body weights were reduced in rats receiving 25 mg/kg PCB while thymus weights were decreased in rats receiving 10 and 25 mg/kg. NK cell activity was reduced in rats dosed for 15 weeks at 10 and 25 mg/kg. The LP response to phytohemagglutinin was enhanced in rats dosed for 15 weeks at 25 mg/kg PCB. Exposure of rats to PCB did not affect the MLR or CTL responses. Other groups of rats were exposed to cyclophosphamide (CY) and served as positive controls for the immune assays employed. CY induced alterations in all of the immune parameters measured, indicating that this is a sensitive battery of immune function tests which is capable of detecting immune alterations in the rat.

  4. Assessment of the immunotoxic potential of the fungicide dinocap in mice

    Energy Technology Data Exchange (ETDEWEB)

    Smialowicz, R.J.; Luebke, R.W.; Riddle, M.M.

    1992-01-01

    The immunotoxic potential of dinocap was evaluated in female C57BL/6J mice following in vivo and in vitro exposure to the fungicide. In in vivo studies, groups of mice were dosed with technical grade dinocap at dosages ranging from 12.5 to 50 mg/kg/d and selected immune functions examined. Twelve days of dosing with dinocap at 25 mg/kg/d resulted in decreased thymus weights and cellularity, and increased spleen weights. Lymphoproliferative responses to concanavalin A (Con A) and phytohemagglutinin (PHA) were reduced in thymocytes from mice dosed at 25 mg/kg/d dinocap. The cytotoxic T lymphocyte (CTL) response to P815 mastocytoma cells was enhanced in mice exposed for 7 days to 25 mg/kg/d dinocap. In vitro studies using murine thymocytes cultured with dinocap (10 ug/ml for 72 hr) resulted in suppression of the proliferative response to Con A and PHA. These results suggest that dinocap is immunotoxic in the mouse, causing effects on T lymphocytes.

  5. [Plant lectins and embryonic tissue factors as probes for studying the mechanisms of neural induction in amphibians].

    Science.gov (United States)

    Mikhaĭlov, A T; Gorgoliuk, N A

    1992-01-01

    Using various experimental techniques, we have demonstrated that animal pole ectoderm (APE) of Rana temporaria embryos at the stage of early gastrula is a good target tissue for testing the neuralizing (N) factors. In this respect R. temporaria APE is comparable with APE of some other amphibian species. We found that concanavalin A (con A), phytohemagglutinin (PHA) and embryonic brain-derived neuralizing factor (EBDNF; a factor extracted from the chick embryonic brain and partially purified) have a pronounced N-effect on the APE of R. temporaria. In order to analyse possible mechanisms of N-action of these factors, we have cultured APE explants for 3 or 18 h in the medium containing various concentrations of con A, PHA of EBDNF. All these factors could produce neuralization in 50% explants. However, the optimal concentration and time of exposure were different. This is an evidence for different mechanisms of reception and transmission of a N-signal in each particular case. It appears that the APE consists of several cell subpopulations which differ in their threshold sensitivity to the N-effect of studied agents.

  6. Emotional Freedom Technique (EFT Effects on Psychoimmunological Factors of Chemically Pulmonary Injured Veterans.

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    Abdolreza Babamahmoodi

    2015-02-01

    Full Text Available Emotional Freedom Technique (EFT as a new therapeutic technique in energy psychology has positive effects on psychological and physiological symptoms, and quality of life. In this research we studied the effect of this treatment on immunological factors. This study tested whether 8-week group sessions of EFT (compared to a wait-list control group with emphasis on patient's respiratory, psychological and immunological problems in chemically pulmonary injured veterans (N=28 can affect on immunological and psychological factors. Mixed effect linear models indicated that EFT improved mental health (F=79.24, p=0 and health-related quality of life (F=13.89, p=0.001, decreased somatic symptoms (F=5.81, p=0.02, anxiety/insomnia (F=24.03, p<0.001, social dysfunction (F=21.59, p<0.001, frequency and severity of respiratory symptoms (F=20.38, p<0.001, and increased lymphocyte proliferation with nonspecific mitogens Concanavalin A (Con A (F=14.32, p=0.001 and Phytohemagglutinin (PHA (F=12.35, p=0.002, and peripheral blood IL-17 (F=9.11, p=0.006. This study provides an initial indication that EFT may be a new therapeutic approach for improving psychological and immunological factors.

  7. Umbilical Cord Tissue-Derived Mesenchymal Stem Cells Induce T Lymphocyte Apoptosis and Cell Cycle Arrest by Expression of Indoleamine 2, 3-Dioxygenase

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    Xiuying Li

    2016-01-01

    Full Text Available It has been reported that human mesenchymal stem cells are able to inhibit T lymphocyte activation; however, the discrepancy among different sources of MSCs is not well documented. In this study, we have compared the MSCs from bone marrow (BM, adipose tissue (AT, placenta (PL, and umbilical cord (UC to determine which one displayed the most efficient immunosuppressive effects on phytohemagglutinin-induced T cell proliferation. Among them we found that hUC-MSC has the strongest effects on inhibiting T cell proliferation and is chosen to do the further study. We observed that T lymphocyte spontaneously released abundant IFN-γ. And IFN-γ secreted by T lymphocyte could induce the expression of indoleamine 2, 3-dioxygenase (IDO in hUC-MSCs. IDO was previously reported to induce T lymphocyte apoptosis and cell cycle arrest in S phase. When cocultured with hUC-MSCs, T lymphocyte expression of caspase 3 was significantly increased, while Bcl2 and CDK4 mRNA expression decreased dramatically. Addition of 1-methyl tryptophan (1-MT, an IDO inhibitor, restored T lymphocyte proliferation, reduced apoptosis, and induced resumption of the cell cycle. In addition, the changes in caspase 3, CDK4, and Bcl2 expression were reversed by 1-MT. These findings demonstrate that hUC-MSCs induce T lymphocyte apoptosis and cell cycle arrest by expressing abundant IDO and provide an explanation for some of the immunomodulatory effects of MSCs.

  8. Adaptive response in human blood lymphocytes exposed to non-ionizing radiofrequency fields: resistance to ionizing radiation-induced damage.

    Science.gov (United States)

    Sannino, Anna; Zeni, Olga; Romeo, Stefania; Massa, Rita; Gialanella, Giancarlo; Grossi, Gianfranco; Manti, Lorenzo; Vijayalaxmi; Scarfì, Maria Rosaria

    2014-03-01

    The aim of this preliminary investigation was to assess whether human peripheral blood lymphocytes which have been pre-exposed to non-ionizing radiofrequency fields exhibit an adaptive response (AR) by resisting the induction of genetic damage from subsequent exposure to ionizing radiation. Peripheral blood lymphocytes from four healthy donors were stimulated with phytohemagglutinin for 24 h and then exposed for 20 h to 1950 MHz radiofrequency fields (RF, adaptive dose, AD) at an average specific absorption rate of 0.3 W/kg. At 48 h, the cells were subjected to a challenge dose (CD) of 1.0 or 1.5 Gy X-irradiation (XR, challenge dose, CD). After a 72 h total culture period, cells were collected to examine the incidence of micronuclei (MN). There was a significant decrease in the number of MN in lymphocytes exposed to RF + XR (AD + CD) as compared with those subjected to XR alone (CD). These observations thus suggested a RF-induced AR and induction of resistance to subsequent damage from XR. There was variability between the donors in RF-induced AR. The data reported in our earlier investigations also indicated a similar induction of AR in human blood lymphocytes that had been pre-exposed to RF (AD) and subsequently treated with a chemical mutagen, mitomycin C (CD). Since XR and mitomycin-C induce different kinds of lesions in cellular DNA, further studies are required to understand the mechanism(s) involved in the RF-induced adaptive response.

  9. Immunostimulatory acivity of Calophyllum brasiliense, Ipomoea pes-caprae and Matayba elaeagnoides demonstrated by human peripheral blood mononuclear cells proliferation.

    Science.gov (United States)

    Philippi, Marina Elisa; Duarte, Bruna Momm; Da Silva, Carolina Vieira; De Souza, Michel Thomaz; Niero, Rivaldo; Cechinel Filho, Valdir; Bueno, Edneia Casagranda

    2010-01-01

    This study evaluates the effect of methanol extracts of three Brazilian medicinal plants on in vitro proliferation of human mononuclear cells. Lymphoproliferation assay was carried out by incubating human peripheral blood mononuclear cells from healthy donors (1 x 10(6) cells/mL) with extracts of Calophyllum brasiliense (roots), Ipomoea pes-caprae (whole plant) and Matayba elaeagnoides (bark), both at 10, 50, 100 and 200 microg/mL, alone or with phytohemagglutinin (PHA, 5 microg/mL), in 96-well microplates at 37 degrees C with 5% CO2, for 72 h. The quantification of cell proliferation assay was performed by blue tetrazolium (MTT) reduction with reading at 540 nm. Cells incubated with only the culture medium were used as negative control for cell proliferation, while the positive control consisted of cells and PHA. The results suggest that the extracts of all three studied plants induce T lymphocyte proliferation. I. pes-caprae showed immunostimulatory activity three times higher than the C. brasiliense extract, while that of the M. elaeagnoides extract was 1.5 times higher. The results demonstrate immunostimulatory effects of these three plants, therefore the continuity of these studies is recommended, in order to determine the active principles.

  10. Metal accumulation and evaluation of effects in a freshwater turtle.

    Science.gov (United States)

    Yu, Shuangying; Halbrook, Richard S; Sparling, Donald W; Colombo, Robert

    2011-11-01

    A variety of contaminants have been detected in aquatic and terrestrial environments around the Paducah Gaseous Diffusion Plant (PGDP), Kentucky. The presence of these contaminants at the PGDP may pose a risk to biota, yet little is known about the bioaccumulation of contaminants and associated effects in wildlife, especially in aquatic turtles. The current study was initiated to evaluate: (1) the accumulation of heavy metals (Cd, Cr, Cu, Pb, and Hg) in aquatic ecosystems associated with the PGDP using red-eared slider turtle (Trachemys scripta elegans) as biomonitors; (2) maternal transfer of heavy metals; and (3) potential hematological and immunological effects resulting from metal accumulation. A total of 26 turtles were collected from 7 ponds located south, adjacent, and north of the PGDP. Liver Cu concentrations were significantly different among ponds and Cu concentrations in eggs were positively correlated with female Cu concentrations in kidney. The concentrations of heavy metals measured in turtle tissues and eggs were low and, based on previous studies of reptiles and established avian threshold levels of heavy metals, did not appear to have adverse effects on aquatic turtles inhabiting ponds near the PGDP. However, total white blood cell counts, heterophil to lymphocyte ratio, and phytohemagglutinin stimulation index were correlated with metal concentrations. Because other factors may affect the hematological and immunological indices, further investigation is needed to determine if these effects are associated with metal exposure, other contaminants, or disease.

  11. Proliferative kinetics and chromosome damage in trisomy 21 lymphocyte cultures exposed to gamma-rays and bleomycin

    Energy Technology Data Exchange (ETDEWEB)

    Morimoto, K.; Kaneko, T.; Iijima, K.; Koizumi, A.

    1984-04-01

    Lymphocytes from patients with Down's syndrome (trisomy 21) have been investigated for cell cycle kinetics, cell proliferation delays, and chromosomal aberrations after exposure to gamma-rays or bleomycin. Analysis by sister chromatid differential staining revealed that trisomy 21 lymphocytes started cell cycling about 5 hr earlier than did normal diploid lymphocytes after phytohemagglutinin stimulation as a whole, but that cycling trisomic and normal cells had the same mean cell cycle times. When exposed to gamma-rays or bleomycin in G0, trisomy 21 lymphocytes showed a 30% or, on average, 50% longer duration of cell turnover times, respectively, than normal cells; only bleomycin-treated trisomic cells had a biphasic dose-response. Frequencies of dicentrics and rings in first-division cells after gamma-ray or bleomycin exposure were twice as high in trisomic cells as in normal cells. The frequency of aberrations decreased by 50% (gamma-ray-exposed) or 65 to 85% (bleomycin-treated) through successive divisions; trisomic cells showed a more marked decline in aberration yields compared to normal cells after bleomycin treatment. These data support the idea that circulating lymphocytes in trisomy 21 patients have a shorter average life span or a younger average age.

  12. T-lymphocytes from AIDS patients are unable to synthesize ribonucleotides de novo in response to mitogenic stimulation. Impaired pyrimidine responses are already evident at early stages of HIV-1 infection.

    Science.gov (United States)

    Bofill, M; Fairbanks, L D; Ruckemann, K; Lipman, M; Simmonds, H A

    1995-12-15

    Proliferative defects have been reported at the level of DNA synthesis, even in T-lymphocytes from asymptomatic human immunodeficiency virus type-1+ (HIV-1+) patients. Since purine and pyrimidine ribonucleotide availability is crucial for proliferation, we compared the ability of HIV-1- and HIV-1+ T-lymphocytes (> 95% CD4+ and CD8+) to activate de novo biosynthetic and salvage pathways following phytohemagglutinin stimulation using 14C-labeled precursors. The striking abnormality already detectable in asymptomatic patients' cells was the impaired ability of CTP, UDP-Glc, and UTP pools to expand over 72 h (44-70% of control), although ATP and GTP pools and responses were normal. In symptomatic patients, resting T-cells showed markedly reduced pyrimidine pools (53-74% of control) with no change following activation. Relatively normal ATP, GTP, and NAD pools masked the same impaired response of de novo synthesis to activation, with ATP and GTP being reduced by 50% at 48 h. Purine salvage was more active than the control in unstimulated HIV-1+ cells. This impaired de novo synthesis in HIV-1+ T-lymphocytes severely restricts the availability of ribonucleotides for vital growth-related activities such as membrane expansion and strand break repair as well as DNA and RNA synthesis. The data indicate that resting T-lymphocytes from symptomatic patients survive through enhanced salvage, but the stimulation induces metabolic cell death, and provide an explanation for the activation-associated lymphocyte death seen in HIV-1+ T-lymphocytes.

  13. Effects of Euphorbia milii latex on mitogen-induced lymphocyte proliferation

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    I.F. Delgado

    2014-03-01

    Full Text Available The crude latex of "Crown-of-Thorns" (Euphorbia milii var hislopii, syn E.splendens is a potent plant molluscicide. For this reason, toxicological studies have been performed to evaluate the health risks posed by its use in schistosomiasis control programs. The present study is part of a more comprehensive immunotoxicological evaluation of this molluscicide. Here, we investigated the effects of E. milii latex on the proliferation of human lymphocytes in vitro. Lyophilized latex of E. milii (0, 0.5, 5, 25 and 50 µg/ml was incubated with whole blood in the presence of proliferation stimulators, i.e. lectins (phytohemagglutinin, concanavalin A and pokeweed mitogen, as well as with human monoclonal antibody against CD3 and tetanus toxoid. Cell proliferation was measured by ³H-thymidine incorporation, and the effects of latex on mitogen-induced cell proliferation were compared to the effects of 10 ng/ml of 12-O-tetradecanoylphorbol-13-acetate (TPA. Results showed that mitogen-induced cell proliferation was markedly enhanced by E. milii latex. This synergistic effect of latex on mitogen-induced lymphocyte proliferation may be due to the presence of TPA-like phorbol esters and/or to mitogenic plant lectins.

  14. Involvement of the thymus and cellular immune system in craniofacial malformation syndromes.

    Science.gov (United States)

    Scheuerle, A E; Good, R A; Habal, M B

    1990-04-01

    Craniofacial structures, the aortic arch, thymus, and parathyroid glands all arise from the embryologic pharyngeal pouches, and DiGeorge and Job craniofacial malformation syndromes have defined immunologic deficiencies. The question addressed by this study is whether patients with other pharyngeal pouch malformations could also have immunologic abnormalities. Twelve patients, 4 female and 8 male, were selected at random from the Tampa Bay Craniofacial Center. Their diagnoses included: cleft lip/cleft palate, hemifacial microsomia/Goldenhar syndrome, Treacher-Collins syndrome, craniofacial hemangiomata, cranio-synostosis syndromes, and Tessier 13 cleft. Fresh blood samples were analyzed against age-matched controls for immunoglobin number, using immunoelectrophoresis, T-cell, B-cell, and natural killer cell quantity via Coulter counter and monoclonal antibody labeling, as well as lymphocyte stimulation and response functions with phytohemagglutinin, concanavalin A, pokeweed mitogen, and Staphylococcus aureus mitogens. All patients studied had some abnormality of their immune systems. Seven had specific T-cell abnormalities and three patients had abnormalities in all three categories studied. This indicates that patients with any pharyngeal pouch malformation may have an abnormality of the immune system.

  15. [Resting metabolic rate, stress, testosterone, and induced immune response in "spring" and "fall" males of Campbell dwarf hamsters. Rearing under the long day conditions].

    Science.gov (United States)

    Rogovin, K A; Bushuev, A V; Khrushchova, A M; Vasil'eva, N Iu

    2013-01-01

    We have studied morphological and physiological traits of even-young males of Campbell dwarf hamsters (Phodopus campbelli Thomas, 1905) born at the end of summer ("fall males") and at the end of winter ("spring males") in a vivarium with constant 14-hour day length (14D:10N). After removal from parental cages at the age of one month, males were kept in isolation under the same light conditions. The results obained signify the statistical difference between "fall" and "spring" males in resting metabolic rate, morphological traits associated with sexual activity, some endocrine and immunologic characteristics. Spring males had higher resting metabolic rate, higher body mass in the middle of experiment, bigger testes, seminal vesicles, higher concentration of testosterone in blood and more intensive T-cell immune response to the intracutaneous injection of phytohemagglutinin. They did not differ significantly in basal level of blood cortisole and antibodies production in response to sheep red blood cells (SRBC) antigen challenge, but possessed lower adrenocortical response to the social stressor and adrenocorticotropic hormone. GLM analysis showed that cortisol level in blood after 10 min encounter of males in the open arena, and resting metabolic rate were the only factors significantly influenced humoral immune response to SRBC. When intensity of T-cell immune response was considered as dependent variable, season turned out to be the only factor in the final model that caused a significant effect.

  16. Lamprey buccal gland secretory protein-2 (BGSP-2 inhibits human T lymphocyte proliferation

    Directory of Open Access Journals (Sweden)

    Jing SUN, Shuiyan YU, Zhuang XUE, Cenjie LIU, Yu WU, Xin LIU, Qingwei LI

    2010-04-01

    Full Text Available Lamprey is a representative of the agnathans, the most ancient class of vertebrates. Parasitic lampreys secrete anticoagulant from their buccal glands and prevent blood coagulation of host fishes. We identified a buccal gland secretory protein-2 (BGSP-2 from a buccal gland cDNA library of Lampetra japonica. The full-length BGSP-2 gene was cloned and the recombinant BGSP-2 protein was generated. The role of BGSP-2 on lymphocyte proliferation was studied by examining its effects on human T lymphocytes. We found that lamprey BGSP-2 was able to effectively block the proliferation of T cells in vitro by inducing G1/S cell cycle arrest. Furthermore, it inhibited the proliferation of human T lymphocytes stimulated by phytohemagglutinin (PHA at a minimum concentration of 0.1μg/ml. Our data suggest that lamprey BGSP-2 is able to block the mitosis of human T lymphocytes at the G1/S point, and has the potential of anti-proliferative effect on PHA-activated T lymphocytes [Current Zoology 56 (2: 252–258, 2010].

  17. The effect of Jeo Dang-Tang on cytokines production in the patients with cerebral infarction.

    Science.gov (United States)

    Jeong, Hyun-Ja; Kang, Sei-Young; Kim, Sang-Yong; Lee, Sang-Gwan; Lee, Sung-Geun; Sung, Kang-Keyng; Kim, Hyung-Min

    2003-11-01

    The herbal formulation "Jeo Dang-Tang" (JDT) has long been used for various cerebrovascular diseases. However, very little has scientific investigation been carried out. The aim of the present study is to investigate the effect of JDT on the production of various cytokines in the patients with cerebral infarction (CI). Peripheral blood mononuclear cells (PBMC) obtained from the patients with CI were cultured for 24h in the presence or absence of lipopolysaccharide (LPS) or phytohemagglutinin (PHA). The amount of interleukin (IL)-4, IL-10 and transforming growth factor (TGF)-1beta, in culture supernatant, was significantly increased in the JDT, LPS or PHA treated cells compared to unstimulated cells (P < 0.05). We also show that increased IL-4, and IL-10 level by LPS or PHA was significantly inhibited by JDT in a dose-dependent manner. Maximal inhibition rate of IL-4 and IL-10 production by JDT was 45 +/- 2% and 51 +/- 5% for LPS-stimulated cell and 41.5 +/- 3% and 70.8 +/- 2% for PHA-stimulated cells, respectively (P < 0.05). On the other hand, JDT significantly increased the LPS or PHA-induced TGF-beta1 production (P < 0.05). These data suggest that JDT has a regulatory effect on the cytokines production, which might explain its beneficial effect in the treatment of CI.

  18. Effects of recombinant human GM-CSF on proliferation of clonogenic cells in acute myeloblastic leukemia.

    Science.gov (United States)

    Griffin, J D; Young, D; Herrmann, F; Wiper, D; Wagner, K; Sabbath, K D

    1986-05-01

    Proliferation of acute myeloblastic leukemia (AML) cells in vitro is limited in most cases to a small subset of blasts that have several properties of stem cells. These leukemic colony-forming cells (AML-CFU) generally require addition of exogenous growth factors for proliferation in agar or methylcellulose. These factors can be supplied by media conditioned by phytohemagglutinin-stimulated normal leukocytes or by CSF-secreting tumor cell lines. However, the exact factor or factors required for stimulation of AML-CFU growth have not been defined. We compared the AML-CFU stimulatory activity of a human recombinant GM-CSF with that of GCT-CM, Mo-CM, and the PHA-leukocyte feeder system in 15 cases of AML. In each of the 12 cases that required exogenous growth factors for maximum AML-CFU growth, recombinant GM-CSF could replace either GM-CSF or Mo-CM, and could partially replace the PHA-leukocyte feeder system. These results indicate that this GM-CSF is a growth promoter of AML-CFU in these culture systems.

  19. Influence of phthalates on in vitro innate and adaptive immune responses.

    Directory of Open Access Journals (Sweden)

    Juliana Frohnert Hansen

    Full Text Available Phthalates are a group of endocrine disrupting chemicals, suspected to influence the immune system. The aim of this study was to investigate the influence of phthalates on cytokine secretion from human peripheral blood mononuclear cells. Escherichia coli lipopolysaccharide and phytohemagglutinin-P were used for stimulation of monocytes/macrophages and T cells, respectively. Cells were exposed for 20 to 22 hours to either di-ethyl, di-n-butyl or mono-n-butyl phthalate at two different concentrations. Both diesters were metabolised to their respective monoester and influenced cytokine secretion from both monocytes/macrophages and T cells in a similar pattern: the secretion of interleukin (IL-6, IL-10 and the chemokine CXCL8 by monocytes/macrophages was enhanced, while tumour necrosis factor (TNF-α secretion by monocytes/macrophages was impaired, as was the secretion of IL-2 and IL-4, TNF-α and interferon-γ by T cells. The investigated phthalate monoester also influenced cytokine secretion from monocytes/macrophages similar to that of the diesters. In T cells, however, the effect of the monoester was different compared to the diesters. The influence of the phthalates on the cytokine secretion did not seem to be a result of cell death. Thus, results indicate that both human innate and adaptive immunity is influenced in vitro by phthalates, and that phthalates therefore may affect cell differentiation and regenerative and inflammatory processes in vivo.

  20. Influence of phthalates on in vitro innate and adaptive immune responses.

    Science.gov (United States)

    Hansen, Juliana Frohnert; Nielsen, Claus Henrik; Brorson, Marianne Møller; Frederiksen, Hanne; Hartoft-Nielsen, Marie-Louise; Rasmussen, Åse Krogh; Bendtzen, Klaus; Feldt-Rasmussen, Ulla

    2015-01-01

    Phthalates are a group of endocrine disrupting chemicals, suspected to influence the immune system. The aim of this study was to investigate the influence of phthalates on cytokine secretion from human peripheral blood mononuclear cells. Escherichia coli lipopolysaccharide and phytohemagglutinin-P were used for stimulation of monocytes/macrophages and T cells, respectively. Cells were exposed for 20 to 22 hours to either di-ethyl, di-n-butyl or mono-n-butyl phthalate at two different concentrations. Both diesters were metabolised to their respective monoester and influenced cytokine secretion from both monocytes/macrophages and T cells in a similar pattern: the secretion of interleukin (IL)-6, IL-10 and the chemokine CXCL8 by monocytes/macrophages was enhanced, while tumour necrosis factor (TNF)-α secretion by monocytes/macrophages was impaired, as was the secretion of IL-2 and IL-4, TNF-α and interferon-γ by T cells. The investigated phthalate monoester also influenced cytokine secretion from monocytes/macrophages similar to that of the diesters. In T cells, however, the effect of the monoester was different compared to the diesters. The influence of the phthalates on the cytokine secretion did not seem to be a result of cell death. Thus, results indicate that both human innate and adaptive immunity is influenced in vitro by phthalates, and that phthalates therefore may affect cell differentiation and regenerative and inflammatory processes in vivo.

  1. The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in G0 and non-random in G1 human lymphocytes.

    Science.gov (United States)

    Hulspas, R; Houtsmuller, A B; Krijtenburg, P J; Bauman, J G; Nanninga, N

    1994-07-01

    The nuclear topography of pericentromeric DNA of chromosome 11 was analyzed in G0 (nonstimulated) and G1 [phytohemagglutinin (PHA) stimulated] human lymphocytes by confocal microscopy. In addition to the nuclear center, the centrosome was used as a second point of reference in the three-dimensional (3D) analysis. Pericentromeric DNA of chromosome 11 and the centrosome were labeled using a combination of fluorescent in situ hybridization (FISH) and immunofluorescence. To preserve the 3D morphology of the cells, these techniques were performed on whole cells in suspension. Three-dimensional images of the cells were analyzed with a recently developed 3D software program (Interactive Measurement of Axes and Positioning in 3 Dimensions). The distribution of the chromosome 11 centromeres appeared to be random during the G0 stage but clearly non-random during the G1 stage, when the nuclear center was used as a reference point. Further statistical analysis of the G1 cells revealed that the centromeres were randomly distributed in a shell underlying the nuclear membrane. A topographical relationship between the centrosome and the centromeres appeared to be absent during the G0 and G1 stages of the cell cycle.

  2. A new flow cytometric method for quantitative assessment of lymphocyte mitogenic potentials.

    Science.gov (United States)

    Yamamura, Y; Rodriguez, N; Schwartz, A; Eylar, E; Bagwell, B; Yano, N

    1995-01-01

    A new flow cytometric method was developed to quantitatively assess lymphocyte proliferation simultaneously for different subsets. The cells were stained with a fluorescent dye, PKH-26 and were stimulated with mitogens. The fluorescence intensities (FL2) of proliferating cells were measured by flow cytometry; and each subset was identified by the use of a monoclonal antibody (Mab)-fluorescein-isothiocyanate (FITC) (FL1). FL2 histograms were then analyzed by the cell proliferation model based on the ModFit software (Verity). This new method revealed information which could not be obtained by conventional mitogen assays. For example, the CD4+ and the CD4- T-subsets responded to phytohemagglutinin (PHA) quite differently from each other and it was indicated that activation of one population could significantly alter the response of the other. In addition, even within a subset, all activated cells did not proliferate uniformly. Some cells divided only once while others underwent further cellular division during the same time period. The method is, therefore, invaluable for studying the nature and the extent of interactions between different cellular subsets within a culture.

  3. An immunological assessment of patients with anorexia nervosa.

    Science.gov (United States)

    Golla, J A; Larson, L A; Anderson, C F; Lucas, A R; Wilson, W R; Tomasi, T B

    1981-12-01

    Patients with most forms of protein-calorie malnutrition are typically more susceptible to infection. We studied the immunological consequences of a subgroup of malnourished subjects--nine patients with anorexia nervosa, who typically have a lower incidence of infection. The profiles of the patients with anorexia nervosa deviated from the reported typical profile of significantly depressed cell-mediated immunity in subjects with more common forms of protein-calorie malnutrition, demonstrating normal T-lymphocyte populations and unimpaired proliferative lymphocyte responsiveness to mitogenic stimulation with phytohemagglutinin and concanavalin A. In fact, mitogen responsiveness was significantly elevated above that of controls, and with nutritional repletion, this enhanced responsiveness regressed toward control values. Since impaired cell-mediated immunity has been consistently documented in other malnourished populations, and presumably contributes to their increased propensity toward infection, the maintenance of a relatively intact cell-mediated immune system may be an important factor separating the malnourished anorexia nervosa patient from other protein-calorie malnourished patients.

  4. Selective Regulation of Human Immunodeficiency Virus-Infected CD4+ Lymphocytes by a Synthetic Immunomodulator Leads to Potent Virus Suppression In Vitro and in hu-PBL-SCID Mice

    Science.gov (United States)

    Bahr, George M.; Darcissac, Edith C. A.; Castéran, Nathalie; Amiel, Corinne; Cocude, Cécile; Truong, Marie-José; Dewulf, Joëlle; Capron, André; Mouton, Yves

    2001-01-01

    We have previously observed that the synthetic immunomodulator Murabutide inhibits human immunodeficiency virus type 1 (HIV-1) replication at multiple levels in macrophages and dendritic cells. The present study was designed to profile the activity of Murabutide on CD8-depleted phytohemagglutinin-activated lymphocytes from HIV-1-infected subjects and on the outcome of HIV-1 infection in severe combined immunodeficiency mice reconstituted with human peripheral blood leukocytes (hu-PBL-SCID mice). Maintaining cultures of CD8-depleted blasts from 36 patients in the presence of Murabutide produced dramatically reduced levels of viral p24 protein in the supernatants. This activity correlated with reduced viral transcripts and proviral DNA, was evident in cultures harboring R5, X4-R5, or X4 HIV-1 isolates, was not linked to inhibition of cellular DNA synthesis, and did not correlate with β-chemokine release. Moreover, c-myc mRNA expression was down-regulated in Murabutide-treated cells, suggesting potential interference of the immunomodulator with the nuclear transport of viral preintegration complexes. On the other hand, daily treatment of HIV-1-infected hu-PBL-SCID mice with Murabutide significantly reduced the viral loads in plasma and the proviral DNA content in human peritoneal cells. These results are the first to demonstrate that a clinically acceptable synthetic immunomodulator with an ability to enhance the host's nonspecific immune defense mechanisms against infections can directly regulate cellular factors in infected lymphocytes, leading to controlled HIV-1 replication. PMID:11435574

  5. Immuno-allergological properties of aluminium oxide (Al2O3) ceramics and nickel sulfate in humans.

    Science.gov (United States)

    Thomas, P; Barnstorf, S; Summer, B; Willmann, G; Przybilla, B

    2003-03-01

    For more than 30 years aluminium oxide (Al(2)O(3)) ceramics have been used for implants in maxillofacial and orthopaedic surgery. Up to now, there are no reports and also no investigations on hypersensitivity reactions. In order to evaluate the aspects of immuno-allergological reactivity to aluminium oxide ceramics, skin testing and in vitro lymphocyte activation studies were performed. The patch test reactivity to a standard series of contact allergens and to an aluminium oxide (Al(2)O(3)) disk was examined in a consecutive series of 250 patients frequenting a University Dermatology Clinic. Furthermore, peripheral blood mononuclear cells (PBMC) of 15 nickel-allergic and 15 non-allergic individuals were cultured with medium alone, with the pan T-cell mitogen phytohemagglutinine (PHA) or with nickel sulfate (NiSO(4)). By additional presence/absence of Al(2)O(3) disk its influence on the cytokine secretion pattern and proliferative response was investigated. The results show that in contrast to a high frequency of delayed-type hypersensitivity to standard contact allergens, no allergic skin reactions to Al(2)O(3) ceramics occured. The IL-4 and IFN-gamma production in vitro remained almost unchanged by the presence of Al(2)O(3) disk as well as the proliferative response of PBMC of non-allergic individuals. Cellular reactivity of nickel allergic and non-allergic donors was partly enhanced upon contact to Al(2)O(3) disks.

  6. Studies on T-cell colony formation in chronic renal failure (CRF) patients.

    Science.gov (United States)

    Wakabayashi, Y; Sugimoto, M; Ishiyama, T; Horie, S; Abe, S; Hirose, S; Okuda, T

    1989-12-01

    In order to study the possibility of abnormal differentiation and proliferation of T-cell precursors in chronic renal failure (CRF), we studied T-cell colony formation in CRF patients. The two-step monolayer method, with phytohemagglutinin-P as the inducer, was used for T-cell colony formation. In our results, colony formation was markedly reduced in CRF patients in comparison with normal controls, with about half of the former showing no colony growth. All cases showed a significant increase in colony numbers with in vitro plasmapheresis (the replacement of autologous plasma in the culture system with normal AB plasma). A significant increase in colony numbers was also seen with the addition of exogenous interleukin-2 (IL-2). The addition of IL-2 in the presence of normal plasma, in particular, induced an increase in colony numbers to near the levels in normal subjects. These results suggest that T-cell precursors exist in near normal numbers in CRF patients and that there are uremic inhibitors in the plasma. A reduced production of IL-2 is also indicated. These factors may be involved in the pathogenesis of immunodeficiency in CRF patients.

  7. The approaches in detecting cell cycle specificity of Fas-mediated apoptosis in leukemia cell lines and activated PBLs in vitro%体外Fas介导细胞凋亡的细胞周期特异性的检测方法

    Institute of Scientific and Technical Information of China (English)

    何小军; 胡静; 李小兰

    2006-01-01

    Objective: To establish a system in detecting the cell cycle specificity induced by recombinant human Fas ligand in vitro,so as to provide a reliable platform for further exploring the mechanism of cell cycle control and regulation in Fas-mediated apoptosis.Methods: The target cells-leukaemia cell lines and activated peripheral blood lymphocytes stimulated by phytohemagglutinin were incubated with recombinant human Fas ligand for 6 to 36 h,apoptosis was detected by sub-G1,common annexin-V/PI and modified annexin V and propidium iodide (API) methods and analysed by flow cytometry.Results: The modified API method demonstrated that Fas-mediated apoptosis was cell cycle specific and initiated at G1 phase.The common annexinV/PI method showed the most appropriate condition for the detection of typical cell cycle-specific apoptosis.The sub-G1 method could only illuminate late apoptosis and DNA histogram.Conclusion: Fas-mediated apoptosis was cell cycle-specific and initiated at G1 phase.Based on the modified APl and common AnnexinV/PI methods,the establishment of stable and typical cell cycle-specific model in Fas-mediated apoptosis in vitro was feasible.

  8. Sustained improvement of intractable rheumatoid arthritis after total lymphoid irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Field, E.H.; Strober, S.; Hoppe, R.T.; Calin, A.; Engleman, E.G.; Kotzin, B.L.; Tanay, A.S.; Calin, H.J.; Terrell, C.P.; Kaplan, H.S.

    1983-08-01

    Total lymphoid irradiation (TLI) was administered to 11 patients who had intractable rheumatoid arthritis that was unresponsive to conventional medical therapy, including aspirin, multiple nonsteroidal antiinflammatory drugs, gold salts, and D-penicillamine. Total lymphoid irradiation was given as an alternative to cytotoxic drugs such as azathioprine and cyclophosphamide. After radiotherapy, 9 of the 11 patients showed a marked improvement in clinical disease activity as measured by morning stiffness, joint tenderness, joint swelling, and overall functional abilities. The mean improvement of disease activity in all patients ranged from 40-70 percent and has persisted throughout a 13-28 month followup period. This improvement permitted the mean daily steroid dose to be reduced by 54%. Complications included severe fatigue and other constitutional symptoms during radiotherapy, development of Felty's syndrome in 1 patient, and an exacerbation of rheumatoid lung disease in another. After therapy, all patients exhibited a profound T lymphocytopenia, and a reversal in their T suppressor/cytotoxic cell to helper cell ratio. The proliferative responses of peripheral blood mononuclear cells to phytohemagglutinin, concanavalin A, and allogeneic leukocytes (mixed leukocyte reaction) were markedly reduced, as was in vitro immunoglobulin synthesis after stimulation with pokeweed mitogen. Alterations in T cell numbers and function persisted during the entire followup period, except that the mixed leukocyte reaction showed a tendency to return to normal values.

  9. Enhanced DNA repair, immune function and reduced toxicity of C-MED-100, a novel aqueous extract from Uncaria tomentosa.

    Science.gov (United States)

    Sheng, Y; Bryngelsson, C; Pero, R W

    2000-02-01

    Female W/Fu rats were gavaged daily with a water-soluble extract (C-MED-100) of Uncaria tomentosa supplied commercially by CampaMed at the doses of 0, 5, 10, 20, 40 and 80 mg/kg for 8 consecutive weeks. Phytohemagglutinin (PHA) stimulated lymphocyte proliferation was significantly increased in splenocytes of rats treated at the doses of 40 and 80 mg/kg. White blood cells (WBC) from the C-MED-100 treatment groups of 40 and 80 mg/kg for 8 weeks or 160 mg/kg for 4 weeks were significantly elevated compared with controls (P tomentosa extracts at the doses of 10-80 mg/kg for 8 weeks or 160 mg/kg for 4 weeks, no acute or chronic toxicity signs were observed symptomatically. In addition, no body weight, food consumption, organ weight and kidney, liver, spleen, and heart pathological changes were found to be associated with C-MED-100 treatment.

  10. A comparative study of physicochemical characteristics and functionalities of pinto bean protein isolate (PBPI) against the soybean protein isolate (SPI) after the extraction optimisation.

    Science.gov (United States)

    Tan, Ee-San; Ying-Yuan, Ngoh; Gan, Chee-Yuen

    2014-01-01

    Optimisation of protein extraction yield from pinto bean was investigated using response surface methodology. The maximum protein yield of 54.8 mg/g was obtained with the optimal conditions of: temperature=25 °C, time=1 h and buffer-to-sample ratio=20 ml/g. PBPI was found to obtain high amount of essential amino acids such as leucine, lysine, and phenylalanine compared to SPI. The predominant proteins of PBPI were vicilin and phytohemagglutinins whereas the predominant proteins of SPI were glycinin and conglycinins. Significantly higher emulsifying capacity was found in PBPI (84.8%) compared to SPI (61.9%). Different isoelectric points were found in both PBPI (4.0-5.5) and SPI (4.0-5.0). Also, it was found that PBPI obtained a much higher denaturation temperature of 110.2 °C compared to SPI (92.5 °C). Other properties such as structural information, gelling capacity, water- and oil-holding capacities, emulsion stability as well as digestibility were also reported.

  11. Chromosomal abnormalities in infertile men referred to iran blood transfusion organization research center.

    Science.gov (United States)

    Frouzandeh, Mahjoubi; Saeideh, Soleimani; Sanaz, Mantegy

    2010-10-01

    The prevalence of somatic chromosomal abnormalities in infertile male individuals has been reported to vary in different literatures. The aim of this study was to investigate the frequency of chromosomal aberrations among infertile men referred to the Cytogenetic Laboratory of Iran Blood Transfusion Organization Research Centre (IBTO). Chromosomal analysis was performed on phytohemag-glutinin (PHA)-stimulated peripheral lymphocyte cultures of 1052 infertile men using standard cytogenetic methods. The study took place during 1997 to 2007. Total chromosome alterations were revealed in 161 (15.30%) infertile men. The most prevalent chromosomal abnormality in the infertile men was 47, XXY, that was seen in 94 (58.38%) men while one of them had a mosaic karyotype: mos 47, XX[54]/47,XXY[18]/46,XY[9]. In 37 (22.98%) cases, structural aberrations were detected. There were 30 (18.63%) cases of sex reversal. Cytogenetic studies of these patients showed increased chromosomal abnormalities in infertile men in comparison with that of the normal population, justifying the need for cytogenetic analysis of men with idiopathic infertility.

  12. Effect of spirulina on the secretion of cytokines from peripheral blood mononuclear cells.

    Science.gov (United States)

    Mao, T K; VAN DE Water, J; Gershwin, M E

    2000-01-01

    ABSTRACT The purpose of this study was to evaluate the immunomodulatory activity of Spirulina, a bluegreen alga used as a food supplement. The effects of Spirulina on the secretion of three cytokines from unstimulated and stimulated human peripheral blood mononuclear cells (PBMC) were examined. In resting PBMC, Spirulina stimulated secretion of interleukin (IL)-1beta, IL-4, and interferon (IFN)-gamma to nearly 2.0, 3.3, and 13.6 times basal levels, respectively. Spirulina induced levels of IFN-gamma (229 +/- 104 pg/ml) that were comparable to those seen after phytohemagglutinin (PHA) stimulation (476 +/- 121 pg/ml). However, it was much less mitogenic than PHA (13.1 +/- 6.9 pg/ml) with respect to the induction of IL-4 secretion (0.34 +/- 0.1 pg/ml). In PHA-stimulated cells, Spirulina enhanced secretion of IL-1beta, IL-4, and IFN-beta by 2.9, 4.0., and 1.6 times, respectively. Although Spirulina stimulates several cytokines, it is clearly more effective in the generation of a Thl-type response. This in vitro study offers additional data for consideration of the potential therapeutic benefits of Spirulina.

  13. Effects of a Spirulina-based dietary supplement on cytokine production from allergic rhinitis patients.

    Science.gov (United States)

    Mao, T K; Van de Water, J; Gershwin, M E

    2005-01-01

    Spirulina represents a blue-green alga that is widely produced and commercialized as a dietary supplement for modulating immune functions, as well as ameliorating a variety of diseases. We have previously shown that the in vitro culture of Spirulina with human peripheral blood mononuclear cells (PBMCs) modulated the production of cytokines. In the present study, we evaluated the impact of a Spirulina-based dietary supplement (Earthrise Nutritionals, Inc., Irvine, CA) on patients with allergic rhinitis by assessing the production of cytokines [interleukin (IL)-4, interferon (IFN)-gamma, and IL-2] critical in regulating immunoglobulin E-mediated allergy. In a randomized double-blinded crossover study versus placebo, allergic individuals were fed daily with either placebo or Spirulina, at 1,000 mg or 2,000 mg, for 12 weeks. PBMCs isolated before and after the Spirulina feeding were stimulated with phytohemagglutinin (PHA) prior to determining the levels of cytokine from cell culture supernatants. Although Spirulina seemed to be ineffective at modulating the secretion of Th1 cytokines (IFN-gamma and IL-2), we discovered that Spirulina, administered at 2,000 mg/day, significantly reduced IL-4 levels by 32% from PHA-stimulated cells. These results indicate that Spirulina can modulate the Th profile in patients with allergic rhinitis by suppressing the differentiation of Th2 cells mediated, in part, by inhibiting the production of IL-4. To our knowledge, this is the first human feeding study that demonstrates the protective effects of Spirulina towards allergic rhinitis.

  14. Deoxyribonucleic acid damage study in primary amenorrhea by comet assay and karyotyping

    Directory of Open Access Journals (Sweden)

    Sarah Ramamurthy

    2013-01-01

    Full Text Available Aim: This study aims at evaluating the chromosomal abnormalities and deoxyribonucleic acid (DNA damage in cases with primary amenorrhea by karyotyping and comet assay. Study Design: A total of 30 cases of primary amenorrhea were recruited. Secondary sexual characters were assessed by Tanner staging. Chromosomal analysis was performed by conventional phytohemagglutinin stimulated lymphocyte cell culture technique. Alkaline version of comet assay was used to evaluate DNA damage. Results: The chromosomal pattern of 20 subjects (66.7% was found to be normal (46,XX. Two subjects had 46,XY pattern and eight subjects had Turner syndrome (45,X or 45,X/46,XX. The comet parameters were found to be increased among subjects with 45,X monosomy, when compared to the rest of the study group and also in subjects with Tanner stage 1 when compared to stage 2. Conclusion: Comet assay revealed increased DNA damage in cases with 45,X monosomy, compared with subjects with 46,XX and 46,XY karyotype, which correlated with clinical features.

  15. Lack of genotoxic effects (micronucleus induction) in human lymphocytes exposed in vitro to 900 MHz electromagnetic fields.

    Science.gov (United States)

    Zeni, O; Chiavoni, A S; Sannino, A; Antolini, A; Forigo, D; Bersani, F; Scarfì, M R

    2003-08-01

    In the present study, we investigated the induction of genotoxic effects in human peripheral blood lymphocytes after exposure to electromagnetic fields used in mobile communication systems (frequency 900 MHz). For this purpose, the incidence of micronuclei was evaluated by applying the cytokinesis-block micronucleus assay. Cytotoxicity was also investigated using the cytokinesis-block proliferation index. The experiments were performed on peripheral blood from 20 healthy donors, and several conditions were tested by varying the duration of exposure, the specific absorption rate (SAR), and the signal [continuous-wave (CW) or GSM (Global System of Mobile Communication) modulated signal]. The following exposures were carried out: (1) CW intermittent exposure (SAR = 1.6 W/kg) for 6 min followed by a 3-h pause (14 on/off cycles); (2) GSM signal, intermittent exposure as described in (1); (3) GSM signal, intermittent exposure as described in (1) 24 h before stimulation with phytohemagglutinin (8 on/off cycles); (4) GSM signal, intermittent exposure (SAR = 0.2 W/kg) 1 h per day for 3 days. The SARs were estimated numerically. No statistically significant differences were detected in any case in terms of either micronucleus frequency or cell cycle kinetics.

  16. Phytochemical screening of the dichloromethane–ethanolic extract of Eriosema campestre var. macrophylum roots and its antiproliferative effect on human peripheral blood lymphocytes

    Directory of Open Access Journals (Sweden)

    Michaelle G. Santos

    Full Text Available ABSTRACT Eriosema campestre var. macrophylum (Grear Fortunato, Fabaceae, is a native plant of the Brazilian Cerrado and the decoction of its roots has been used by folk medicine for the therapy of inflammatory diseases. In this study we aimed to investigate the effect of the dichloromethane–ethanolic extract of E. campestre roots on the proliferative response of lymphocytes and to examine the profile of IL-2 production. The effect of dichloromethane–ethanolic extract of E. campestre on the proliferation of phytohemagglutinin-stimulated lymphocytes was evaluated by using flow cytometry and the cell supernatants were assayed for IL-2 concentrations by using an enzyme-linked immunosorbent assay. The phytochemical screening of E. campestre roots was performed to determine the main secondary metabolites through chromogenic and precipitation reactions and by using HPLC-PAD. In addition to the presence of subclasses of flavonoids (flavones and flavonols in dichloromethane–ethanolic extract of E. campestre, we observed that the extract induced a concentration-dependent decrease in IL-2 levels on the supernatant of the cell cultures as well as an antiproliferative effect on T lymphocytes, including CD4+ and CD8+ cells. The anti-inflammatory effects attributed to E. campestre by folk medicine may partly be explained by its antiproliferative action on T lymphocytes.

  17. Ocular toxoplasmosis in immunosuppressed nonhuman primates

    Energy Technology Data Exchange (ETDEWEB)

    Holland, G.N.; O' Connor, G.R.; Diaz, R.F.; Minasi, P.; Wara, W.M.

    1988-06-01

    To investigate the role of cellular immunodeficiency in recurrent toxoplasmic retinochoroiditis, six Cynomolgus monkeys (Macaca fascicularis) with healed toxoplasmic lesions of the retina were immunosuppressed by total lymphoid irradiation. Three months prior to irradiation 30,000 Toxoplasma gondii organisms of the Beverley strain had been inoculated onto the macula of eye in each monkey via a pars plana approach. Toxoplasmic retinochoroiditis developed in each animal, and lesions were allowed to heal without treatment. During total lymphoid irradiation animals received 2000 centigrays (cGy) over a 7-week period. Irradiation resulted in an immediate drop in total lymphocyte counts and decreased ability to stimulate lymphocytes by phytohemagglutinin. Weekly ophthalmoscopic examinations following irradiation failed to show evidence of recurrent ocular disease despite persistent immunodeficiency. Four months after irradiation live organisms were reinoculated onto the nasal retina of the same eye in each animal. Retinochoroidal lesions identical to those seen in primary disease developed in five of six animals. Toxoplasma organisms therefore were able to proliferate in ocular tissue following the administration of immunosuppressive therapy. This study fails to support the hypothesis that cellular immunodeficiency alone will initiate recurrent toxoplasmic retinochoroiditis. Results suggest that reactivation of disease from encysted organisms involves factors other than suppression of Toxoplasma proliferation. If reactivation occurs by other mechanisms, however, cellular immunodeficiency then may allow development of extensive disease.

  18. Comparison of the immunosuppressive effect of fractionated total lymphoid irradiation (TLI) vs conventional immunosuppression (CI) in renal cadaveric allotransplantation

    Energy Technology Data Exchange (ETDEWEB)

    Waer, M.; Vanrenterghem, Y.; Ang, K.K.; van der Schueren, E.; Michielsen, P.; Vandeputte, M.

    1984-02-01

    Beginning in November 1981, eight patients with end stage diabetic nephropathy underwent renal cadaveric transplantation after TLI. Transplantation was done between 2 to 11 days after the end of a fractionated TLI to a total dose of 20 to 30 Gy. During the same observation period, 60 nondiabetic patients with end stage renal disease of different origin also received a cadaveric kidney graft, with a conventional regimen of immunosuppression that consists of anti-lymphocyte-globulin, tapering high doses of prednisone, and azathioprine. Phytohemagglutinin (PHA)-, concanavalin A (con A)-, and pokeweed mitogen (PWM)-induced blastogenesis, as well as the mixed lymphocyte reaction (MLR) and the cell-mediated lympholysis (CML) decreased progressively during the first months after conventional immunosuppression to 50% of the pretransplantation level, and remained there for the first year after transplantation. These tests were much more impaired after TLI and again no recovery occurred during the first year. In the clinic, the more profound immunosuppression in TLI patients was more frequently associated with viral infections (cytomegalovirus and herpes zoster). The incidence of rejections, however, was somewhat less frequent in the TLI-treated group and occurred significantly later. After TLI, the mean cumulative dose of steroids needed for kidney transplantation during the first year after transplantation could be substantially reduced.

  19. Avidin inhibits PHA-induced human peripheral blood mononuclear cell proliferation

    Directory of Open Access Journals (Sweden)

    Cicia Firakania

    2016-04-01

    Full Text Available Background: Cell proliferation occurs not only in normal but also in cancer cells. Most of cell proliferation inhibition can be done by inhibiting the DNA synthesis, notably by intervening the formation of purine or pyrimidine. In purine de novo synthesis, it was assumed that biotin plays a role as a coenzyme in carboxylation reaction, one of the pivotal steps in the purine de novo pathways. The aim of this study was to see the avidin potency to bind biotin and inhibit mitosis.Methods: Peripheral blood mononuclear cell (PBMC was cultured in RPMI-1640 medium and stimulated by phytohemagglutinin (PHA in the presence or absence of interleukin-2 (IL-2, with or without avidin. The effect of avidin addition was observed at 24, 48, and 72 hours for cell proliferation, viability, and cell cycle. Statistical analysis was done by one-way ANOVA.Results: Avidin inhibited cell proliferation and viability in culture under stimulation by PHA with and without IL-2. Cell cycle analysis showed that avidin arrested the progression of PBMC after 72 hours of culture. Most cells were found in G0/G1 phase.Conclusion: Inhibition of biotin utilization by avidin binding can halt cell proliferation.

  20. Laser scanning fluorescence microscopic measurement of the movement of cleaving egg surface of Rana Amurensis

    Institute of Scientific and Technical Information of China (English)

    GUGUOYAN; ChengtangXu; 等

    1995-01-01

    By laser scanning fluorescence microscopy for quantitative measurement of fluorescence intensity changes on egg surface stained with fluorescein isothiocyanate during cleavage furrow extending forward,it was found that in area of presumptive cleavage furrow the scanning curve became ∨ shape,indicating dark stripe appeared in that place.Then the fluorescence intensity increased at the place where the bottom of ∨ shape had located,and the scanning curve turned to ∧ shape,indicating single stripe was formed.While enhanced fluorescence appeared on the borders of ∧ shape,an M shape curve was found,showing double stripe occurred.During the distance between two borders of M shape incresing from 50μm to 100μm,a fluorescence peak came to sight in the middle of the M shape,which being the cleavge furrow bottom.The two lateral sides of furrow bottom with decreasing fluorescence were nascent membrane.At that time the curve became W shape.By the sides of cleavage furrow the the stress folds became conspicous after double stripe stage,showing the stretching of the egg surface being increased.With our[31,33]and others[32] reports that polylysine could induce the appearance of nascent membrane and phytohemagglutinins could decrease or prevent the appearance of nascent membrane,we believed the idea of Schroeder[25] that increasing mechanical stress could initiate nascent membrane formation and thought that the stresslay to the outsides of cleavage furrow.

  1. State-dependent physiological maintenance in a long-lived ectotherm, the painted turtle (Chrysemys picta).

    Science.gov (United States)

    Schwanz, Lisa; Warner, Daniel A; McGaugh, Suzanne; Di Terlizzi, Roberta; Bronikowski, Anne

    2011-01-01

    Energy allocation among somatic maintenance, reproduction and growth varies not only among species, but among individuals according to states such as age, sex and season. Little research has been conducted on the somatic (physiological) maintenance of long-lived organisms, particularly ectotherms such as reptiles. In this study, we examined sex differences and age- and season-related variation in immune function and DNA repair efficiency in a long-lived reptile, the painted turtle (Chrysemys picta). Immune components tended to be depressed during hibernation, in winter, compared with autumn or spring. Increased heterophil count during hibernation provided the only support for winter immunoenhancement. In juvenile and adult turtles, we found little evidence for senescence in physiological maintenance, consistent with predictions for long-lived organisms. Among immune components, swelling in response to phytohemagglutinin (PHA) and control injection increased with age, whereas basophil count decreased with age. Hatchling turtles had reduced basophil counts and natural antibodies, indicative of an immature immune system, but demonstrated higher DNA repair efficiency than older turtles. Reproductively mature turtles had reduced lymphocytes compared with juvenile turtles in the spring, presumably driven by a trade-off between maintenance and reproduction. Sex had little influence on physiological maintenance. These results suggest that components of physiological maintenance are modulated differentially according to individual state and highlight the need for more research on the multiple components of physiological maintenance in animals of variable states.

  2. In vitro immune functions in thiamine-replete and -depleted lake trout (Salvelinus namaycush)

    Science.gov (United States)

    Ottinger, Christopher A.; Honeyfield, Dale C.; Densmore, Christine L.; Iwanowicz, Luke R.

    2014-01-01

    In this study we examined the impacts of in vivo thiamine deficiency on lake trout leukocyte function measured in vitro. When compared outside the context of individual-specific thiamine concentrations no significant differences were observed in leukocyte bactericidal activity or in concanavalin A (Con A), and phytohemagglutinin-P (PHA-P) stimulated leukocyte proliferation. Placing immune functions into context with the ratio of in vivo liver thiamine monophosphate (TMP – biologically inactive form) to thiamine pyrophosphate (TPP – biologically active form) proved to be the best indicator of thiamine depletion impacts as determined using regression modeling. These observed relationships indicated differential effects on the immune measures with bactericidal activity exhibiting an inverse relationship with TMP to TPP ratios, Con A stimulated mitogenesis exhibiting a positive relationship with TMP to TPP ratios and PHA-P stimulated mitogenesis exhibiting no significant relationships. In addition, these relationships showed considerable complexity which included the consistent observation of a thiamine-replete subgroup with characteristics similar to those seen in the leukocytes from thiamine-depleted fish. When considered together, our observations indicate that lake trout leukocytes experience cell-type specific impacts as well as an altered physiologic environment when confronted with a thiamine-limited state.

  3. Tobacco plants transformed with the bean. alpha. ai gene express an inhibitor of insect. alpha. -amylase in their seeds. [Nicotiana tabacum; Tenebrio molitor

    Energy Technology Data Exchange (ETDEWEB)

    Altabella, T.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (USA))

    1990-06-01

    Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant {alpha}-amylases. We recently presented strong circumstantial evidence that this {alpha}-amylase inhibitor ({alpha}Al) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5{prime} and 3{prime} flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M{sub r} 10,000-18,000) that cross-react with antibodies to the bean {alpha}-amylase inhibitor. Most of these polypeptides bind to a pig pancreas {alpha}-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas {alpha}-amylase activity as well as the {alpha}-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called {alpha}ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera.

  4. Effects of Malnutrition on Children's Immunity to Bacterial Antigens in Northern Senegal

    Science.gov (United States)

    Gaayeb, Lobna; Sarr, Jean B.; Cames, Cecile; Pinçon, Claire; Hanon, Jean-Baptiste; Ndiath, Mamadou O.; Seck, Modou; Herbert, Fabien; Sagna, Andre B.; Schacht, Anne-Marie; Remoue, Franck; Riveau, Gilles; Hermann, Emmanuel

    2014-01-01

    To evaluate immunity to vaccine-preventable diseases according to nutritional status, a longitudinal study was conducted in Senegalese children ages 1–9 years old. A linear regression analysis predicted that weight for age was positively associated with immunoglobulin G (IgG) response to tetanus toxoid in children born during the rainy season or at the beginning of the dry season. A relationship between village, time of visits, and levels of antibodies to tetanus showed that environmental factors played a role in modulating humoral immunity to tetanus vaccine over time. Moreover, a whole-blood stimulation assay highlighted that the production of interferon-γ (IFN-γ) in response to tetanus toxoid was compromised in stunted children. However, the absence of cytokine modulation in response to Mycobacterium tuberculosis-purified protein derivatives and phytohemagglutinin suggests that the overall ability to produce IFN-γ was preserved in stunted children. Therefore, these results show that nutritional status can specifically alter the efficacy of long-lasting immunity to tetanus. PMID:24445198

  5. Genetic difference in the proliferative response to T mitogens between Hi/PHA and Lo/PHA lymphocytes is independent of accessory cell function.

    Science.gov (United States)

    Stiffel, C; Liacopoulos-Briot, M; Decreusefond, C; Parlebas, J

    1987-01-01

    The role of the macrophage as accessory cell in the proliferative response of lymphocytes to phytohemagglutinin (PHA) was studied in two lines of mice genetically selected for high and low responsiveness to T mitogens. Adherent cell depletion of lymph node cells abrogated the low (Lo)/PHA response, but only partially inhibited the high (Hi)/PHA response. Addition of peritoneal cells provided either by Hi/PHA or by Lo/PHA mice equally restored Hi/PHA responsiveness but had only a slight reconstituting effect on the inhibited Lo/PHA response. Equivalent enhancement or suppression of proliferation of untreated lymph node cells was obtained by the addition of increasing percentages of each of the two peritoneal cell populations. However, the maximum level of the Lo/PHA response never reached that of Hi/PHA cells. These data indicate that the bidirectional selective breeding has not modified the potentialities of the macrophages as accessory cells but has resulted in an impaired response of Lo/PHA lymphocytes to the signals delivered either by accessory cells or by T mitogens.

  6. Responding to inflammatory challenges is less costly for a successful avian invader, the house sparrow (Passer domesticus), than its less-invasive congener.

    Science.gov (United States)

    Lee, Kelly A; Martin, Lynn B; Wikelski, Martin C

    2005-09-01

    When introduced into new regions, invading organisms leave many native pathogens behind and also encounter evolutionarily novel disease threats. In the presence of predominantly novel pathogens that have not co-evolved to avoid inducing a strong host immune response, costly and potentially dangerous defenses such as the systemic inflammatory response could become more harmful than protective to the host. We therefore hypothesized that introduced populations exhibiting dampened inflammatory responses will tend to be more invasive. To provide initial data to assess this hypothesis, we measured metabolic, locomotor, and reproductive responses to inflammatory challenges in North American populations of the highly invasive house sparrow (Passer domesticus) and its less-invasive relative, the tree sparrow (Passer montanus). In the house sparrow, there was no effect of phytohemagglutinin (PHA) challenge on metabolic rate, and there were no detectable differences in locomotor activity between lipopolysaccharide (LPS)-injected birds and saline-injected controls. In contrast, tree sparrows injected with PHA had metabolic rates 20-25% lower than controls, and LPS injection resulted in a 35% drop in locomotor activity. In a common garden captive breeding experiment, there was no effect of killed-bacteria injections on reproduction in the house sparrow, while tree sparrows challenged with bacteria decreased egg production by 40% compared to saline-injected controls. These results provide some of the first data correlating variation in immune defenses with invasion success in introduced-vertebrate populations.

  7. Evolutionary analysis of the APA genes in the Phaseolus genus: wild and cultivated bean species as sources of lectin-related resistance factors?

    Science.gov (United States)

    Lioi, L; Galasso, I; Lanave, C; Daminati, M G; Bollini, R; Sparvoli, F

    2007-11-01

    The APA (Arcelin/Phytohemagglutinin/alpha-Amylase inhibitor) gene family is composed of various members, present in Phaseolus species and coding for lectin and lectin-related seed proteins having the double role of storage and defense proteins. Here members of the APA family have been identified by immunological, functional, and molecular analyses and representative genes were sequenced in nine wild species of Phaseolus. All taxa possessed at least one member of the true lectin gene. No arcelin type sequences have been isolated from the species examined. Among the wild species studied, only P. costaricensis contained an alpha-amylase inhibitor (alpha-AI). In addition P. augusti, P. maculatus, P. microcarpus, and P. oligospermus showed the presence of the lectin-related alpha-amylase inhibitor-like (AIL) genes and alpha-AI activity. Data from Southern blot analysis indicated the presence of only one lectin gene in P. parvulus and P. filiformis, while an extensive gene duplication of the APA locus was found in the other Phaseolus species. Phylogenetic analysis carried out on the nucleotide sequences showed the existence of two main clusters and clearly indicated that lectin-related genes originated from a paralogous duplication event preceding the development of the ancestor to the Phaseolus genus. The finding of detectable alpha-AI activity in species containing AIL genes suggests that exploiting APA genes variability in the Phaseolus genus may represent a valuable tool to find new members that may have acquired insecticidal activities.

  8. Effects of different levels of vitamin premix in finisher diets on performance, immuno - competence and meat lipid oxidation of chickens fed on corn - soybean meal

    Directory of Open Access Journals (Sweden)

    Hoseein Moravej

    2013-03-01

    Full Text Available The present study was carried out to examine the effects of a vitamin premix (VPreduction or withdrawal from finisher diet (29-43 days on performance,immuno-competence,and characteristicsof leg bones and meat lipid oxidation of chickens fed oncorn-soybeanmeal based diet. A total of 900 male broiler chickens (Ross 308 were allocatedtofivetreatment groups(0, 33%, 66%, 100% and 133% VP, withninereplicates per treatmentgroup. At 29 and 36 days of ages, four birds from each replicate were injected with sheepredblood cells (SRBC. The cell-mediated immunity was determined via phytohemagglutinin(PHA and 1-chloro 2-4-dinitrobenzen (DNCBat 34 and 42 days of ages.At 33, 38 and 43days of age, 42 days of ages, and two birds of each replicate were slaughteredand boneparameters measured. The oxidative stability was evaluated by thiobarbituric acid reactivesubstances (TBARS on the thigh samples that were stored for 90 day at-80 ̊C. The resultsshowed that reduction or withdrawal of VP from diets at different time points of the finisherperiod did not affect performance, immunocompetence and characteristics of leg bones.Results of TBARS showed thatlipid peroxidation of the treatment without VP wassignificantly higher than of the other treatments when slaughtered at 43 days of age. Finally,the results of this study demonstrated that it is not possible to reduce the VP in finisherbroilers’ diets without negative effects on meat quality during the time of freezing.

  9. The in vitro effects of artificial and natural sweeteners on the immune system using whole blood culture assays.

    Science.gov (United States)

    Rahiman, F; Pool, E J

    2014-01-01

    This article investigates the effects of commercially available artificial (aspartame, saccharin, sucralose) and natural sweeteners (brown sugar, white sugar, molasses) on the immune system. Human whole blood cultures were incubated with various sweeteners and stimulated in vitro with either phytohemagglutinin or endotoxin. Harvested supernatants were screened for cytotoxicity and cytokine release. Results showed that none of the artificial or natural sweeteners proved to be cytotoxic, indicating that no cell death was induced in vitro. The natural sweetener, sugar cane molasses (10 ug/mL), enhanced levels of the inflammatory biomarker IL-6 while all artificial sweeteners (10 ug/mL) revealed a suppressive effect on IL-6 secretion (P sweeteners under stimulatory conditions reduced levels of the biomarker of humoral immunity, Interleukin-10 (P < 0.001). The cumulative suppression of Interleukin-6 and Interleukin-10 levels induced by sucralose may contribute to the inability in mounting an effective humoral response when posed with an exogenous threat.

  10. Monitoring of chimerism using fluorescence in situ hybridization in a child with severe combined immune deficiency following bone marrow transplant

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    Wenger, S.L.; Chen, X.O.; Katz, A.J. [Children`s Hospital of Pittsburgh, PA (United States)]|[Univ. of Pittsburgh, PA (United States)

    1994-09-01

    A boy with severe combined immunodeficiency received a bone marrow transplant from his sister when he was approximately 3 years of age. His peripheral blood karyotype at age 3 and 4 years was 46,XX (20 cells analyzed). Because of a decline in antibody production at 19 years of age, the patient`s peripheral blood was analyzed again for suspected chimerism. His karyotype in phytohemagglutinin (PHA)-stimulated culture was 46,XX in 49 cells and 46,XY in one cell. Both metaphase and interphase cells were examined for sex chromosome constitution using X and Y dual-color alpha-satellite probes for fluorescence in situ hybridization (FISH). FISH results for metaphase cells showed 1/50 XY cells, but 38% of interphase cells showed the presence of both X and Y centromere. Pokeweed mitogen (PWM)-stimulated cultures grew poorly and were therefore analyzed using FISH only: 81% of interphase cells were 46,XX. The discrepancy between metaphase and interphase in the PHA-stimulated cultures most likely represents a failure of this boy`s own XY T-cells to be stimulated.

  11. Genetic and environmental components of phenotypic variation in immune response and body size of a colonial bird, Delichon urbica (the house martin).

    Science.gov (United States)

    Christe, P; Moller, A P; Saino, N; De Lope, F

    2000-07-01

    Directional selection for parasite resistance is often intense in highly social host species. Using a partial cross-fostering experiment we studied environmental and genetic variation in immune response and morphology in a highly colonial bird species, the house martin (Delichon urbica). We manipulated intensity of infestation of house martin nests by the haematophagous parasitic house martin bug Oeciacus hirundinis either by spraying nests with a weak pesticide or by inoculating them with 50 bugs. Parasitism significantly affected tarsus length, T cell response, immunoglobulin and leucocyte concentrations. We found evidence of strong environmental effects on nestling body mass, body condition, wing length and tarsus length, and evidence of significant additive genetic variance for wing length and haematocrit. We found significant environmental variance, but no significant additive genetic variance in immune response parameters such as T cell response to the antigenic phytohemagglutinin, immunoglobulins, and relative and absolute numbers of leucocytes. Environmental variances were generally greater than additive genetic variances, and the low heritabilities of phenotypic traits were mainly a consequence of large environmental variances and small additive genetic variances. Hence, highly social bird species such as the house martin, which are subject to intense selection by parasites, have a limited scope for immediate microevolutionary response to selection because of low heritabilities, but also a limited scope for long-term response to selection because evolvability as indicated by small additive genetic coefficients of variation is weak.

  12. Cytokines and dysregulation of the immune response in human malaria

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    M. Fátima C. Alves

    1992-01-01

    Full Text Available The dysregulation of the immune response by malaria parasite has been considered as a possible constraint to the effectiveness of malaria vaccination. In spite of the important role interleukin-I (IL-1 in malaria are lacking. We found that only 2 out of 35 subjectswith acute malaria showed increased levels of serum IL-1 alpha by enzyme immunoassay. To assess whether IL-1 could interfere with T- lymphocyte responses, blood mononuclear cells from patients infected with Plasmodium falciparum, P. vivax, or healthy subjects were cultured with phytohemagglutinin, and lymphocyte proliferation measured 72h later by 3H-thymidine incorporation. Our data showed that T-lymphocyte responses are depressed both in P. falciparum (10,500 ñ 2,900 and P. vivax malaria (13,000 ñ 3,300, as compared to that of healthy individuals (27,000 ñ 3,000. Addition of IL-1 partially reserved depression of malaria lymphocytes, but had no effect on normal cells. On the other hand, T-lymphocytes from malaria infected-subjects presented a minimal decrease in proliferation, when cultured in the presence of exogenous PGE2. These data indicate the occurrence of two defects of immunoregulation in malaria: a deficiency of IL-1 production by monocytes/macrophages, and an increased resistance of lymphocytes to the antiproliferative effect of PGE2.

  13. Partial impairment of immune functions in peripheral blood leukocytes from aged men with Down's syndrome.

    Science.gov (United States)

    Park, E; Alberti, J; Mehta, P; Dalton, A; Sersen, E; Schuller-Levis, G

    2000-04-01

    Down's syndrome (DS) has been considered a model of accelerated aging and of Alzheimer's disease. We investigated immunologic functions using peripheral blood leukocytes in order to correlate the production of cytokines and development of neuropathological changes of Alzheimer type in aged persons with DS. Cytokine production (IL-1beta, IL-2, IL-6, IL-8, and TNF-alpha), phytohemagglutinin (PHA)-stimulated proliferation of nonadherent monocytes, and superoxide anion production from polymorphonuclear leukocytes were measured. PHA-stimulated proliferation in aged individuals (>30 years old) with DS was significantly lower than that of age- and sex-matched controls (DS vs control, 55,707+/-5810 vs 88,310+/-6994 cpm, P < 0.001). PHA-stimulated IL-2 production was also significantly decreased in aged individuals with DS (DS vs control, 7.1+/-2.1 vs 10.7+/-1.3 ng/ml). Interestingly, the decrease of proliferation and IL-2 production in aged males with DS is significantly greater than in aged women with DS. PHA-stimulated proliferation and IL-2 production of nonadherent monocytes in females was not significantly reduced. IL-1beta production by LPS-activated adherent monocytes was significantly decreased in older adults with DS compared with non-DS controls. Other immune parameters measured in DS were not significantly different from that of age-matched controls. We conclude that there is partial impairment of T lymphocytes in aged persons with DS that is significantly greater in males than in females.

  14. Activation-induced apoptosis in peripheral blood mononuclear cells during hepatosplenic Schistosoma mansoni infections.

    Science.gov (United States)

    Ghoneim, H M; Demian, S R; Heshmat, M G; Ismail, N S; El-Sayed, Laila H

    2008-01-01

    It is well established that programmed cell death (apoptosis) is an important regulator of host responses during infection with a variety of intra- and extra-cellular pathogens. The present work aimed at assessment of in vitro spontaneous and phytohemagglutinin (PHA)-induced apoptosis in mononuclear cells isolated from patients with hepatosplenic form of S. mansoni infections. Cell death data were correlated to the degree of lymphoproliferative responses to PHA as well as to the serum anti-schistosomal antibody titers. A markedly significant increase in PHA-induced apoptosis in lymphocytes isolated from S. mansoni-infected patients was seen when compared to the corresponding healthy controls. However, a slight difference was recorded between the two studied groups regarding the spontaneous apoptosis. This was accompanied with a significant impairment of in vitro PHA-induced lymphoproliferation of T cells from S. mansoni patients. Data of the present study supports the hypothesis that activation-induced cell death (AICD) is a potentially contributing factor in T helper (Th) cell regulation during chronic stages of schistosomiasis, which represents a critically determinant factor in the host-parasite interaction and might influence the destiny of parasitic infections either towards establishment of chronic infection or towards host death.

  15. A citometria de fluxo como instrumento de avaliação da atividade imunomodulatória de extratos e substâncias isoladas de plantas medicinais Flow cytometry as instrument for evaluating the immunopotential of medicinal plant extracts and their isolated compounds

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    Julio C. Machado Junior

    2006-12-01

    Full Text Available A citometria de fluxo vem se consolidando como metodologia para vários estudos de atividade celular e, nesse trabalho, ela foi empregada para avaliar as ações da fitohemaglutinina, dos alcalóides vimblastina e vindolina e dos extratos de Chamomilla recutita (L. Rauschert, Bauhinia microstachya (Raddi Machr. e Himatanthus lancifolius (Muell. Arg. Woodson, conhecidos popularmente no Brasil como camomila, escada-de-macaco e agoniada, respectivamente, sobre a imunomodulação de células mononucleares humanas, comparando-se o desempenho obtido com as alterações morfológicas relacionadas à ativação e proliferação dessas células in vitro. Os resultados demonstram que foi possível identificar, pela metodologia proposta, os efeitos proliferativos estimulantes já descritos para a fitohemaglutinina e para o extrato de C. recutita, assim como para o extrato de B. microstachya, observado pela primeira vez nesse trabalho. A conhecida atividade inibitória da vimblastina sobre a proliferação de linfócitos induzida por fitohemaglutinina, em contraste como a ausência de efeitos da vindolina, também puderam ser evidenciadas. Efeito inibitório foi observado para o extrato de H. lancifolius devido à sua ação tóxica sobre o sistema. Os resultados apresentados sugerem que a citometria de fluxo pode ser usada como uma alternativa metodológica quando se deseja investigar a ação de substâncias puras ou de extratos de plantas sobre a imunomodulação de células mononucleares humanas.Flow cytometry has been widely applied for studying several cellular activities. In this work it has been used to evaluate the effects of phytohemagglutinin, vinblastine, and vindoline upon human mononuclear cells immunomodulation. The same protocol was used to investigate the effects of extracts prepared from Chamomilla recutita (L. Rauschert, Bauhinia microstachya (Raddi Machr., and Himatanthus lancifolius (Muell. Arg. Woodson, plants popularly known in Brazil

  16. Immunomodulatory Changes Induced By High Doses Of Dextromethorphan In Male Rats

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    Hanan Mostafa Rabei

    2013-04-01

    Full Text Available Background:Dextromethorphan (DXM is a synthetic opioid analogue, similar to codeine that has antitussive effect but no opiate-like analgesic activity. It is widely used as an over-the-counter cough suppressant available in various cough and cold preparations; it is one of the often overlooked types of substance abuse by adolescents and young adults in the United States and around the world. So, the present study aims to investigate the side effects of the DXM abuse (sub-chronic and lethal doses on the immune functions in rats. Material and Methods:The rats were divided into three equal groups, the first one served as control, and the second and third were treated. Treated groups received oral doses of DXM which increasing per 10 days (double dose for a month. ResultsWe examined the sub-chronic and lethal effects of DXM administration on the cellular immune responses in rats. T cell stimulator, Phytohemagglutinin showed a significant suppress on lymphocytes of peripheral blood proliferation and a highly significant decrease on phagocytic and killing of S. aureus by PMN and macrophage cells. Moreover, it induced a significant decrease in serum IL-6, and IFN-γ levels, but, it exhibited a highly significant increase in serum IL-10 level throughout the period of experiment. In addition, it induced also a significant decrease in the production of cortisol during the experimental time except the last period of treatment in the 3rd group, where, serum cortisol level gradually return to normal level. Conclusion:These results suggest that sub-chronic and lethal doses of DXM administration in rats disturbed cellular immune responses, exhibited potent anti-inflammatory actions, suppressed of leukocytes dependent production of cytokines such as IL-6 and IFN-γ. Moreover, it has some effects on serum cortisol concentration presumably via blockade of NMDA receptors

  17. [Effect of a non-steroidal anti-inflammatory agent, tolmetin sodium on exudative inflammation in experimental animals (author's transl)].

    Science.gov (United States)

    Nakamura, H; Yokoyama, Y; Motoyoshi, S; Ishii, K; Shimizu, M

    1979-07-01

    Effect of tolmetin sodium(Tol) on acute and subacute exudative inflammation was tested in experimental animals. Tol had a potent inhibitory activity (ED50 = 0.75 mg/kg, p.o.) on the increased vascular permeability induced by acetic acid in mice, and the potency was about 0.4 times that of indomethacin (Ind), and 6-93 times that of ibuprofen (Ibu), phenylbutazone(Phe) and aspirin(Asp). The inhibitory activity of Tol(ED50 = 18.2 mg/kg, p.o.) on UV-induced erythema in guinea pigs was about 0.3 times that of Ind. A recovery of the hind paw edema of rats, produced by a mixture of kaolin and carrageenin, was promoted by oral administration of Tol(2.5 approximately 20 mg/kg x 5/2 days). Tol(80 mg/kg/day, p.o.) showed a significant activity in inhibiting the exudation caused by croton oil in rats, and the activity was about 0.025 times that of Ind and greater than that of Ibu, Phe and Asp. Tol(100-800 microgram/ml) inhibited in a dose-dependent manner the phytohemagglutinin-induced blast transformation of cultured lymphocytes from rat thymus, as did salicylic acid. In vitro, Tol showed a potent activity similar to that of Ibu and Phe in preventing the denaturation of bovine serum albumin and the lysis of rat erythrocytes. From these results, it is suggested that Tol has a particularly potent inhibitory activity on acute exudative inflammation, and the mode of action may be attributed to a mechanism similar to that seen with other acidic non-steroidal anti-inflammatory drugs.

  18. Human T-lymphotropic Virus Type 1-infected Cells Secrete Exosomes That Contain Tax Protein*

    Science.gov (United States)

    Jaworski, Elizabeth; Narayanan, Aarthi; Van Duyne, Rachel; Shabbeer-Meyering, Shabana; Iordanskiy, Sergey; Saifuddin, Mohammed; Das, Ravi; Afonso, Philippe V.; Sampey, Gavin C.; Chung, Myung; Popratiloff, Anastas; Shrestha, Bindesh; Sehgal, Mohit; Jain, Pooja; Vertes, Akos; Mahieux, Renaud; Kashanchi, Fatah

    2014-01-01

    Human T-lymphotropic virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia and HTLV-1-associated myelopathy/tropical spastic paraparesis. The HTLV-1 transactivator protein Tax controls many critical cellular pathways, including host cell DNA damage response mechanisms, cell cycle progression, and apoptosis. Extracellular vesicles called exosomes play critical roles during pathogenic viral infections as delivery vehicles for host and viral components, including proteins, mRNA, and microRNA. We hypothesized that exosomes derived from HTLV-1-infected cells contain unique host and viral proteins that may contribute to HTLV-1-induced pathogenesis. We found exosomes derived from infected cells to contain Tax protein and proinflammatory mediators as well as viral mRNA transcripts, including Tax, HBZ, and Env. Furthermore, we observed that exosomes released from HTLV-1-infected Tax-expressing cells contributed to enhanced survival of exosome-recipient cells when treated with Fas antibody. This survival was cFLIP-dependent, with Tax showing induction of NF-κB in exosome-recipient cells. Finally, IL-2-dependent CTLL-2 cells that received Tax-containing exosomes were protected from apoptosis through activation of AKT. Similar experiments with primary cultures showed protection and survival of peripheral blood mononuclear cells even in the absence of phytohemagglutinin/IL-2. Surviving cells contained more phosphorylated Rb, consistent with the role of Tax in regulation of the cell cycle. Collectively, these results suggest that exosomes may play an important role in extracellular delivery of functional HTLV-1 proteins and mRNA to recipient cells. PMID:24939845

  19. Generation of human hybridomas producing migration inhibitory factor (MIF) and of murine hybridomas secreting monoclonal antibodies to human MIF.

    Science.gov (United States)

    Weiser, W Y; Remold, H G; David, J R

    1985-01-01

    Human T-cell hybridomas were established by hybridization of concanavalin A (Con A)-stimulated human peripheral blood T lymphocytes with cells from a 6-thioguanine-resistant, aminopterin-sensitive mutant line designated CEM-WH4, derived from the continuously growing human T cell line, CEM. High levels of MIF activity were demonstrated in the supernatants of two hybridoma lines, T-CEMA and T-CEMB but not of CEM-WH4 when stimulated with phorbol myristate acetate and phytohemagglutinin. In comparison, MIF derived from Con A-stimulated peripheral blood mononuclear cells showed 100 times less activity. Upon isoelectrofocusing, MIF activity of T-CEMB was found exclusively between pH 4.6 and 5.3 whereas MIF derived from T-CEMA showed heterogeneity with a major peak of MIF recovered at pH 4.6-5.3 and a minor peak at pH 2.4-3.3. These molecules, however, were all found to have an apparent MW of 68,000 and were resistant to trypsin. Most of these characteristics are in accordance with second day pH 3- and pH 5-MIF derived from peripheral blood mononuclear cells. When spleen cells from BALB/c mice immunized with T-CEMB-MIF were used to fuse with NS-1 mouse myeloma cells, nine hybridomas secreting antibodies to human MIF were obtained. Clone D112 which demonstrated the highest MIF-neutralizing activity was found to neutralize MIF derived from T-CEMA, peripheral blood mononuclear cells, and a T cell line, Mo.

  20. Effects of Neuromedin S on the Proliferation of Splenic Lymphocytes and the Cytokine Secretion by Pulmonary Alveolar Macrophages in Pigs in vitro.

    Science.gov (United States)

    Lin, R; Wang, Q; Qi, B; Huang, Y; Yang, G

    2016-09-01

    Neuromedin S (NMS), a 36-amino acid neuropeptide, has been found to be involved in the regulation of the endocrine activity. It has been also detected in immune tissues in mammals, what suggests that NMS may play an important role in the regulation of immune response. The aim of this study was to demonstrate the presence of NMS receptor 1 (NMU1R) and effect of NMS in pig splenic lymphocytes (SPLs) and pulmonary alveolar macrophages (PAMs). The presence of NMU1R in pig SPLs and PAMs was respectively confirmed by reverse transcription-polymerase chain reaction (RT-PCR), western blot analysis and immunocytochemical methods. Furthermore, SPL proliferation was analyzed using the 3-(4,5)-dimethyl-thiahiazo-(-2-yl)-3,5-di-phenytetrazoliumromide (MTT) method. Additionally, the secretion of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in PAMs was all measured by enzyme-linked immunosorbent assay (ELISA) kits. In the present study, the results of RT-PCR and western blot analysis revealed that NMU1R mRNA and protein were both expressed in pig SPLs and PAMs, and the immunocytochemical investigations further revealed that the positive signal of NMU1R immunoreactivity was observed in plasma membranes of both SPLs and PAMs. In the in vitro study, we found that at concentrations of 0.001-1000 nM NMS alone or combined with lipopolysaccharide or phytohemagglutinin significantly increased SPL proliferation. Application of ELISA method showed that NMS could induce the secretion of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in PAMs. These results suggest that NMS can act as a potently positive pro-inflammatory factor and immunomodulatory agent that affects the immune response of immune cells by combining with its receptor NMU1R.

  1. Impairment of T-regulatory cells in cord blood of atopic mothers.

    Science.gov (United States)

    Schaub, Bianca; Liu, Jing; Höppler, Sabine; Haug, Severine; Sattler, Christine; Lluis, Anna; Illi, Sabina; von Mutius, Erika

    2008-06-01

    Maternal atopy is a strong predictor for the development of childhood allergic diseases. The underlying mechanisms are ill defined, yet regulatory T (Treg) and T(H)17 cells may play a key role potentially shaping the early immune system toward a proallergic or antiallergic immune regulation. We examined T(H)1/T(H)2, Treg, and T(H)17 cell responses to innate (lipid A/peptidoglycan) and mitogen/adaptive (phytohemagglutinin/Dermatophagoides pteronyssinus 1) immune stimulation in cord blood from offspring of atopic/nonatopic mothers. Cord blood mononuclear cells from 161 healthy neonates (59% nonatopic, 41% atopic mothers) were investigated regarding Treg and T(H)17 cells (mRNA/surface markers), suppressive function, and proliferation/cytokine secretion. Cord blood from offspring of atopic mothers showed fewer innate-induced Treg cells (CD4(+)CD25(+)high), lower mRNA expression of associated markers (glucocorticoid-induced tumor necrosis factor receptor-related protein/lymphocyte activation gene 3; P cell function was impaired in mitogen-induced suppression of T effector cells in cord blood of offspring from atopic mothers (P = .03). Furthermore, IL-10 and IFN-gamma secretion were decreased in innate-stimulated cord blood of offspring from atopic mothers (P = .04/.05). Innate-induced IL-17 was independent of maternal atopy and highly correlated with IL-13 secretion. In offspring of atopic mothers, Treg cell numbers, expression, and function were impaired at birth. T(H)17 cells were correlated with T(H)2 cells, independently of maternal atopy.

  2. Dendritic cells loaded with pancreatic Cancer Stem Cells (CSCs lysates induce antitumor immune killing effect in vitro.

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    Tao Yin

    Full Text Available According to the cancer stem cells (CSCs theory, malignant tumors may be heterogeneous in which a small population of CSCs drive the progression of cancer. Because of their intrinsic abilities, CSCs may survive a variety of treatments and then lead to therapeutic resistance and cancer recurrence. Pancreatic CSCs have been reported to be responsible for the malignant behaviors of pancreatic cancer, including suppression of immune protection. Thus, development of immune strategies to eradicate pancreatic CSCs may be of great value for the treatment of pancreatic cancer. In this study, we enriched pancreatic CSCs by culturing Panc-1 cells under sphere-forming conditions. Panc-1 CSCs expressed low levels of HLA-ABC and CD86, as measured by flow cytometry analysis. We further found that the Panc-1 CSCs modulate immunity by inhibiting lymphocyte proliferation which is promoted by phytohemagglutinin (PHA and anti-CD3 monoclonal antibodies. The monocyte derived dendritic cells (DCs were charged with total lysates generated from Panc-1 CSCs obtained from tumor sphere culturing. After co-culturing with lymphocytes at different ratios, the Panc-1 CSCs lysates modified DC effectively promoted lymphocyte proliferation. The activating efficiency reached 72.4% and 74.7% at the ratios of 1∶10 and 1∶20 with lymphocytes. The activated lymphocytes secreted high levels of INF-γ and IL-2, which are strong antitumor cytokines. Moreover, Panc-1 CSCs lysates modified DC induced significant cytotoxic effects of lymphocytes on Panc-1 CSCs and parental Panc-1 cells, respectively, as shown by lactate dehydrogenase (LDH assay. Our study demonstrates that the development of CSCs-based vaccine is a promising strategy for treating pancreatic cancer.

  3. Treatment of NZB/NZW mice with total lymphoid irradiation: long-lasting suppression of disease without generalized immune suppression

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    Kotzin, B.L.; Arndt, R.; Okada, S.; Ward, R.; Thach, A.B.; Strober, S.

    1986-05-01

    We used total lymphoid irradiation (TLI; total dose = 3400 rad) to treat the lupus-like renal disease of 6-mo-old female NZB/NZW mice. Similar to our past studies, this treatment resulted in a marked prolongation of survival, decrease in proteinuria, and decrease in serum anti-DNA antibodies compared with untreated littermate controls. Although there was no evidence of disease recurrence in TLI-treated mice until after 12 mo of age, the in vitro proliferative response to phytohemagglutinin by NZB/NZW spleen cells recovered within 6 wk such that responses were greater than control NZB/NZW animals. A similar recovery and overshoot after TLI were evident in the primary antibody response to the T cell-dependent antigen sheep red blood cells (SRBC). Both the total and IgG anti-SRBC antibody responses after TLI were greater than those of untreated NZB/NZW controls, and were comparable with those of untreated non-autoimmune mice. Despite this increased response to mitogens and antigens after TLI, we noted a decrease in spontaneous splenic IgG-secreting cells and a decrease in IgG but not IgM antinuclear antibody production. Nonspecific suppressor cells of the mixed leukocyte response were detectable in the spleens of NZB/NZW mice early after TLI. However, the disappearance of suppressor cells was not associated with recrudescence of disease activity. Furthermore, transfer of large numbers of spleen cells from TLI-treated NZB/NZW mice did not result in disease suppression in untreated age-matched recipients. In summary, treatment of NZB/NZW mice with TLI results in a prolonged remission in autoimmune disease, which is achieved in the absence of generalized immunosuppression.

  4. Lymphocyte Redox Imbalance and Reduced Proliferation after a Single Session of High Intensity Interval Exercise.

    Science.gov (United States)

    Tossige-Gomes, Rosalina; Costa, Karine Beatriz; Ottone, Vinícius de Oliveira; Magalhães, Flávio de Castro; Amorim, Fabiano Trigueiro; Rocha-Vieira, Etel

    2016-01-01

    This study investigated whether an acute session of high-intensity interval training (HIIT) is sufficient to alter lymphocyte function and redox status. Sixteen young healthy men underwent a HIIT session on a cycloergometer, consisting of eight bouts of 1 min at 90-100% of peak power, with 75 seconds of active recovery at 30 W between bouts. Venous blood was collected before, immediately after, and 30 minutes after the HIIT session. In response to Staphylococcus aureus superantigen B (SEB) stimulation, lymphocyte proliferation decreased and the IL-2 concentration increased after the HIIT session. However, the HIIT session had no effect on lymphocyte proliferation or IL-2 response to phytohemagglutinin stimulation. The HIIT session also induced lymphocyte redox imbalance, characterized by an increase in the concentration of thiobarbituric acid reactive substances and a decrease in the activity of the antioxidant enzyme catalase. Lymphocyte viability was not affected by the HIIT session. The frequencies of CD25+ and CD69+ T helper and B lymphocytes in response to superantigen stimulation were lower after exercise, suggesting that superantigen-induced lymphocyte activation was reduced by HIIT. However, HIIT also led to a reduction in the frequency of CD4+ and CD19+ cells, so the frequencies of CD25+ and CD69+ cells within the CD4 and CD19 cell populations were not affected by HIIT. These data indicate that the reduced lymphocyte proliferation observed after HIIT is not due to reduced early lymphocyte activation by superantigen. Our findings show that an acute HIIT session promotes lymphocyte redox imbalance and reduces lymphocyte proliferation in response to superantigenic, but not to mitogenic stimulation. This observation cannot be explained by alteration of the early lymphocyte activation response to superantigen. The manner in which lymphocyte function modulation by an acute HIIT session can affect individual immunity and susceptibility to infection is important

  5. Improvement in clinical signs and cellular immunity of dogs with visceral leishmaniasis using the immunomodulator P-MAPA.

    Science.gov (United States)

    Santiago, Maria Emília B; Neto, Luiz Silveira; Alexandre, Eduardo Costa; Munari, Danísio Prado; Andrade, Mariana Macedo C; Somenzari, Marcos Arruda; Ciarlini, Paulo César; de Lima, V M F

    2013-09-01

    This study investigated the immunotherapeutic potential of the protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride immuno-modulator (P-MAPA) on canine visceral leishmaniasis. Twenty mongrel dogs presenting clinical symptoms compatible with leishmaniasis and diagnosis confirmed by the detection of anti-leishmania antibodies were studied. Ten dogs received 15 doses of the immunomodulator (2.0 mg/kg) intramuscularly, and 10 received saline as a placebo. Skin and peripheral blood samples were collected following administration of the immunomodulator. The groups were followed to observe for clinical signals of remission; parasite load in the skin biopsies using real-time PCR, the cytokines IL-2, IL-10 and IFN-γ in the supernatant of peripheral blood mononuclear cells stimulated in vitro with either total promastigote antigen or phytohemagglutinin measured by capture ELISA, and changes in CD4⁺ and CD8⁺ T cell subpopulations evaluated by flow cytometry. Comparison between the groups showed that treatment with the immunomodulator promoted improvement in clinical signs and a significant reduction in parasite load in the skin. In peripheral blood mononuclear cell cultures, supernatants showed a decrease in IL-10 levels and an increase in IL-2 and IFN-γ. An increase in CD8⁺ T cells was observed in peripheral blood. In addition, the in vitro leishmanicidal action of P-MAPA was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and no leishmanicidal activity was detected. These findings suggest that P-MAPA has potential as an immunotherapeutic drug in canine visceral leishmaniasis, since it assists in reestablishing partial immunocompetence of infected dogs.

  6. Effect of vitamin E levels on the cell-mediated immunity of broilers vaccinated against coccidiosis

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    ICM da Silva

    2011-03-01

    Full Text Available Studies on the relationships between animal nutrition and immunity have sought reliable methodologies to measure responses. Cell-mediated immune response is similarly studied in humans. The cutaneous basophil hypersensitivity test (CBH is one of the methods to measure that response and consists in the infiltration of inflammatory cells, particularly of lymphocytes and basophils, as result of the application of substances capable of inducing cell proliferation in determined sites, such as wings, wattle, and interdigital space in birds. CBH is considered a simple and fast method and can be applied in birds of different ages. In immunocompetence studies with poultry, phytohemagglutinin-P (PHA-P is a commonly used substance, despite the variability of the response related to the method of application (intradermal injection and the antigens used. In the present experiment, PHA-P was used to observe the cell-mediated immune response of 216 chicks fed three dietary levels of vitamin E from 1 to 36 days of age. All birds were immunologically challenged by vaccination against coccidiosis at three days of age and against Newcastle Disease (NCD at 14 and 30 days of age. At 36 days of age, birds were submitted to the CBH test according to the methodology of Corrier & DeLoach (1990. Birds fed 65mg/kg of vitamin E presented lasting cell reaction (p<0.08, which indicates that this vitamin E level improved cell immune response of birds due to its antioxidant and immunomodulating properties. The use of this vitamin E level can be considered by nutritionists under practical conditions, aiming to improve broiler immunity.

  7. X射线诱发外周血淋巴细胞TCR基因突变研究%X-ray induced TCR gene mutation of peripheral blood lymphocytes

    Institute of Scientific and Technical Information of China (English)

    侯殿俊; 马娅; 刘伟; 乔建维; 卢峰; 商希梅; 李洁清; 封丽

    2009-01-01

    objective To study the TCR gene mutation in peripheral blood lymphocytes induced by X-ray exposure using cultured lymphocytes cloning method.Methods Freshly isolated peripheral lymphocytes from healthy aduh donors were irradiated with X-ray in doses ranging from 0 to 8 Gy and cultured with interleukin2 and phytohemagglutinin for 7 days.The mutant frequencies of TCR gene(TCR MF)were detected by flow cytonletry and the dose response curves were fitted.Results TCR MF increased with the dose going up.An aquadratic polynomial dose response model was fitted.Conclusions TCR gene mutation could which serve as a potential biological dosimeter.It might be applied for the estimation of biological dose in emergency exposure.%目的 用培养法研究X射线诱发的人外周血淋巴细胞TCR基因突变情况.方法 以不同剂量(0~8 Gy)的X射线照射新鲜分离的健康成人外周血淋巴细胞,植物血凝素、白细胞介素2(IL-2)协同刺激培养7 d,流式细胞术检测TCR基因突变频率(TCR MF),并拟合剂量效应关系.结果 随着照射剂量的增加,TCR基因突变频率随之上升,最佳拟合曲线为二次多项式模型.结论 TCR基因突变可作为辐射生物剂量计,用于急性辐射照射生物剂量的估算.

  8. Ratite oils promote keratinocyte cell growth and inhibit leukocyte activation.

    Science.gov (United States)

    Bennett, Darin C; Leung, Gigi; Wang, Eddy; Ma, Sam; Lo, Blanche K K; McElwee, Kevin J; Cheng, Kimberly M

    2015-09-01

    Traditionally, native Australian aborigines have used emu oil for the treatment of inflammation and to accelerate wound healing. Studies on mice suggest that topically applied emu oil may have anti-inflammatory properties and may promote wound healing. We investigated the effects of ratite oils (6 emu, 3 ostrich, 1 rhea) on immortalized human keratinocytes (HaCaT cells) in vitro by culturing the cells in media with oil concentrations of 0%, 0.5%, and 1.0%. Peking duck, tea tree, and olive oils were used as comparative controls. The same oils at 0.5% concentration were evaluated for their influence on peripheral blood mononuclear cell (PBMC) survival over 48 hr and their ability to inhibit IFNγ production in PBMCs activated by phytohemagglutinin (PHA) in ELISpot assays. Compared to no oil control, significantly shorter population doubling time durations were observed for HaCaT cells cultured in emu oil (1.51×faster), ostrich oil (1.46×faster), and rhea oil (1.64×faster). Tea tree oil demonstrated significant antiproliferative activity and olive oil significantly prolonged (1.35×slower) cell population doubling time. In contrast, almost all oils, particularly tea tree oil, significantly reduced PBMC viability. Different oils had different levels of inhibitory effect on IFNγ production with individual emu, ostrich, rhea, and duck oil samples conferring full inhibition. This preliminary investigation suggests that emu oil might promote wound healing by accelerating the growth rate of keratinocytes. Combined with anti-inflammatory properties, ratite oil may serve as a useful component in bandages and ointments for the treatment of wounds and inflammatory skin conditions.

  9. The Glycoprofile Patterns of Endothelial Cells in Usual Interstitial Pneumonia

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    A Barkhordari

    2014-09-01

    Full Text Available [THIS ARTICLE HAS BEEN RETRACTED FOR DUPLICATE PUBLICATION] Background: The pathological classification of cryptogenic fibrosing alveolitis has been a matter of debate and controversy for histopathologists. Objective: To identify and specify the glycotypes of capillary endothelial cells in usual interstitial pneumonia (UIP compared to those found in normal tissue. Methods: Sections of formalin-fixed, paraffin-embedded blocks from 16 cases of UIP were studied by lectin histochemistry with a panel of 27 biotinylated lectins and an avidin-peroxidase revealing system. Results: High expression of several classes of glycan was seen de novo in capillary endothelial cells from patients with UIP including small complex and bi/tri-antennary bisected complex N-linked sequences bolund by Concanavalin A and erythro-phytohemagglutinin, respectively, GalNAca1 residues bound by Helix pomatia and Maclura pomifera agglutinins, and L-fucosylated derivatives of type II glycan chains recognized by Ulex europaeus agglutinin-I. Glycans bound by agglutinins from Lycopersicon esculentum (β1,4GlcNAc and Wisteria floribunda (GalNAc as well as GlcNAc oligomers bound by Phytolacca americana and succinylated Wheat Germ agglutinin were also seen in the capillary endothelial cells of UIP. In contrast, L-fucosylated derivatives of type I glycan chains were absent in cells from cases of UIP when Anguilla anguilla agglutinin was applied, unlike the situation in normal tissue. Conclusion: These results may indicate existence of two distinct populations of endothelial cell in UIP with markedly different patterns of glycosylation, reflecting a pattern of differentiation and angiogenesis, which is not detectable morphologically.

  10. Neuroinflammation induces glial aromatase expression in the uninjured songbird brain

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    Saldanha Colin J

    2011-07-01

    Full Text Available Abstract Background Estrogens from peripheral sources as well as central aromatization are neuroprotective in the vertebrate brain. Under normal conditions, aromatase is only expressed in neurons, however following anoxic/ischemic or mechanical brain injury; aromatase is also found in astroglia. This increased glial aromatization and the consequent estrogen synthesis is neuroprotective and may promote neuronal survival and repair. While the effects of estradiol on neuroprotection are well studied, what induces glial aromatase expression remains unknown. Methods Adult male zebra finches (Taeniopygia guttata were given a penetrating injury to the entopallium. At several timepoints later, expression of aromatase, IL-1β-like, and IL-6-like were examined using immunohisotchemistry. A second set of zebra birds were exposed to phytohemagglutinin (PHA, an inflammatory agent, directly on the dorsal surface of the telencephalon without creating a penetrating injury. Expression of aromatase, IL-1β-like, and IL-6-like were examined using both quantitative real-time polymerase chain reaction to examine mRNA expression and immunohistochemistry to determine cellular expression. Statistical significance was determined using t-test or one-way analysis of variance followed by the Tukey Kramers post hoc test. Results Following injury in the zebra finch brain, cytokine expression occurs prior to aromatase expression. This temporal pattern suggests that cytokines may induce aromatase expression in the damaged zebra finch brain. Furthermore, evoking a neuroinflammatory response characterized by an increase in cytokine expression in the uninjured brain is sufficient to induce glial aromatase expression. Conclusions These studies are among the first to examine a neuroinflammatory response in the songbird brain following mechanical brain injury and to describe a novel neuroimmune signal to initiate aromatase expression in glia.

  11. Immune function is related to adult carotenoid and bile pigment levels, but not to dietary carotenoid access during development, in female mallard ducks.

    Science.gov (United States)

    Butler, Michael W; McGraw, Kevin J

    2013-07-15

    Immune function can be modulated by multiple physiological factors, including nutrition and reproductive state. Because these factors can vary throughout an individual's lifetime as a result of environmental conditions (affecting nutrition) or life-history stage (e.g. entering the adult reproduction stage), we must carefully examine the degree to which developmental versus adult conditions shape performance of the immune system. We investigated how variation in dietary access to carotenoid pigments - a class of molecules with immunostimulatory properties that females deposit into egg yolks - during three different developmental time points affected adult immunological and reproductive traits in female mallard ducks (Anas platyrhynchos). In males and females of other avian species, carotenoid access during development affects carotenoid assimilation ability, adult sexual ornamentation and immune function, while carotenoid access during adulthood can increase immune response and reproductive investment (e.g. egg-laying capacity, biliverdin deposition in eggshells). We failed to detect effects of developmental carotenoid supplementation on adult immune function [phytohemagglutinin-induced cutaneous immune response, antibody production in response to the novel antigen keyhole limpet hemocyanin (KLH), or oxidative burst, assessed by changes in circulating nitric oxide levels], carotenoid-pigmented beak coloration, ovarian development, circulating carotenoid levels or concentration of bile pigments in the gall bladder. However, we did uncover positive relationships between circulating carotenoid levels during adulthood and KLH-specific antibody production, and a negative relationship between biliverdin concentration in bile and KLH-specific antibody production. These results are consistent with the view that adult physiological parameters better predict current immune function than do developmental conditions, and highlight a possible, previously unstudied relationship

  12. Toxic effects of dietary methylmercury on immune system development in nestling American kestrels (Falco sparverius).

    Science.gov (United States)

    Fallacara, Dawn M; Halbrook, Richard S; French, John B

    2011-06-01

    This study evaluated the effects of dietary methylmercury (MeHg) on immune system development in captive-reared nestling American kestrels (Falco sparverius) to determine whether T cell-mediated and antibody-mediated adaptive immunity are targets for MeHg toxicity at environmentally relevant concentrations. Nestlings received various diets, including 0 (control), 0.6, and 3.9 µg/g (dry wt) MeHg for up to 18 d posthatch. Immunotoxicity endpoints included cell-mediated immunity (CMI) using the phytohemagglutinin (PHA) skin-swelling assay and antibody-mediated immune response via the sheep red blood cell (SRBC) hemagglutination assay. T cell- and B cell-dependent histological parameters in the spleen, thymus, and bursa of Fabricius were correlated with the functional assays. For nestlings in the 0.6 and 3.9 µg/g MeHg groups, CMI was suppressed by 73 and 62%, respectively, at 11 d of age. Results of this functional assay were correlated with T cell-dependent components of the spleen and thymus. Dose-dependent lymphoid depletion in spleen tissue directly affected the proliferation of T-lymphocyte populations, insofar as lower stimulation indexes from the PHA assay occurred in nestlings with lower proportions of splenic white pulp and higher THg concentrations. Nestlings in the 3.9 µg/g group also exhibited lymphoid depletion and a lack of macrophage activity in the thymus. Methylmercury did not have a noticeable effect on antibody-mediated immune function or B cell-dependent histological correlates. We conclude that T cell-mediated immunosuppression is the primary target of MeHg toward adaptive immunity in developing kestrels. This study provides evidence that environmentally relevant concentrations of MeHg may compromise immunocompetence in a developing terrestrial predator and raises concern regarding the long-term health effects of kestrels that were exposed to dietary MeHg during early avian development.

  13. Induction and repair of DNA damage measured by the comet assay in human T lymphocytes separated by immunomagnetic cell sorting.

    Science.gov (United States)

    Bausinger, Julia; Speit, Günter

    2014-11-01

    The comet assay is widely used in human biomonitoring to measure DNA damage in whole blood or isolated peripheral blood mononuclear cells (PBMC) as a marker of exposure to genotoxic agents. Cytogenetic assays with phytohemagglutinin (PHA)-stimulated cultured T lymphocytes are also frequently performed in human biomonitoring. Cytogenetic effects (micronuclei, chromosome aberrations, sister chromatid exchanges) may be induced in vivo but also occur ex vivo during the cultivation of lymphocytes as a consequence of DNA damage present in lymphocytes at the time of sampling. To better understand whether DNA damage measured by the comet assay in PBMC is representative for DNA damage in T cells, we comparatively investigated DNA damage and its repair in PBMC and T cells obtained by immunomagnetic cell sorting. PBMC cultures and T cell cultures were exposed to mutagens with different modes of genotoxic action and DNA damage was measured by the comet assay after the end of a 2h exposure and after 18h post-incubation. The mutagens tested were methyl methanesulfonate (MMS), (±)-anti-B[a]P-7,8-dihydrodiol-9,10-epoxide (BPDE), 4-nitroquinoline-1-oxide (4NQO), styrene oxide and potassium bromate. MMS and potassium bromate were also tested by the modified comet assay with formamido pyrimidine glycosylase (FPG) protein. The results indicate that the mutagens tested induce DNA damage in PBMC and T cells in the same range of concentrations and removal of induced DNA lesions occurs to a comparable extent. Based on these results, we conclude that the comet assay with PBMC is suited to predict DNA damage and its removal in T cells.

  14. Effects of Supplemental Zinc in a Wheat-Based Diet on Performance, Intestinal Viscosity, Immune System and Lipid Peroxidation of 21-Day Old Broiler Chickens

    Directory of Open Access Journals (Sweden)

    Dibaiee-nia G

    2017-06-01

    Full Text Available We investigated the effects of a wheat-based diet (WBD supplemented with different levels of zinc on the performance, intestinal viscosity, immune system and lipid peroxidation of broiler chickens. A total of 240 Ross 308 day-old male broiler chicks were weighed and assigned to six dietary treatments with four replicates (floor pens of ten birds per pen. Dietary treatments consisted of a WBD without Zn supplement in mineral premix (control, or with 20, 40, 60, 80, and 100 mg/kg of Zn in the diet. Feed intake, body weight gain, and feed conversion ratio were recorded after 21 days. On day 21, blood serum malondialdehyde concentration, intestinal digesta viscosity, and some internal organs were measured. Antibody titer against Sheep red blood cells (SRBC were measured on days 7 and 14 after injection. For evaluation of cutaneous basophil hypersensitivity (CBH response, on d 20, phytohemagglutinin was injected subcutaneously into toe web and 12 and 24 hrs after injection, the thickness of the web was measured. Supplementation of the WBD with 20, 40, 60, and 80 mg Zn/kg significantly improved feed conversion ratio (P < 0.05. Supplementation of Zn significantly decreased the relative weight of abdominal fat pad as well as jejunal viscosity (P < 0.05. Also, Zn supplementation (at all concentrations except 20 mg/kg significantly decreased serum malondialdehyde concentration (P< 0.05. Anti-SRBC titer was significantly increased by supplementation of the WBD with 20 mg/kg Zn (P < 0.05. Supplementation of the WBD with 40 mg/kg Zn significantly increased CBH response (P < 0.05. Overall, the results of this study indicate the importance of Zn supplementation in WBD for improvement of FCR and physicochemical properties of the intestinal contents. Also, supplementation of Zn in the WBD is effective in enhancing immune system responses and antioxidative defense.

  15. Biomolecule screening for efficient attachment of biofunctionalized microparticles to the zona pellucida of mammalian oocytes and embryos.

    Science.gov (United States)

    Novo, Sergio; Ibáñez, Elena; Barrios, Leonardo; Castell, Onofre; Nogués, Carme

    2013-10-01

    Individual tagging of oocytes and embryos through the attachment of micrometer-sized polysilicon barcodes to their zona pellucida (ZP) is a promising approach to ensure their correct identification and traceability in human assisted reproduction and in animal production programs. To provide barcodes with the capacity of binding to the ZP, they must be first biofunctionalized with a biomolecule capable of binding to the ZP of both oocytes and embryos. The aim of this work was to select, among an anti-ZP2 antibody and the two lectins wheat germ agglutinin (WGA) and phytohemagglutinin-L, the most optimal biomolecule for the eventual biofunctionalization of barcodes, using mouse oocytes and embryos and commercially available microspheres as a model. Despite the anti-ZP2 antibody showed the highest number of binding sites onto the ZP surface, as determined by field emission scanning electron microscopy, the binding of anti-ZP2-biofunctionalized microspheres to the ZP of cultured oocytes and embryos was less robust and less stable than the binding of lectin-biofunctionalized ones. WGA proved to be, among the three candidates tested, the most appropriate biomolecule to biofunctionalize microparticles with the aim to attach them to the ZP of both oocytes and embryos and to maintain them attached through oocyte activation (zona reaction) and in vitro culture up to the blastocyst stage. As saccharides recognized by WGA are highly abundant in the ZP of most mammalian species, WGA-biofuncionalized microparticles would be able to attach to the ZP of oocytes/embryos of species other than the mouse, such as humans and farm animals.

  16. Activation and proliferation signals in primary human T lymphocytes inhibited by ergosterol peroxide isolated from Cordyceps cicadae

    Science.gov (United States)

    Kuo, Y C; Weng, S C; Chou, C J; Chang, T T; Tsai, W J

    2003-01-01

    Effects of ergosterol peroxide (C28H44O3; Cpd 6A) from Cordyceps cicadae on phytohemagglutinin (PHA)-stimulated cell proliferation were studied in primary human T cells. The results showed that Cpd 6A suppressed T-cell proliferation for about 24 h after stimulation with PHA. Cell cycle analysis indicated that Cpd 6A arrested the cell cycle progression of activated T cells from the G1 transition to the S phase. To localize the point in the cell cycle where arrest occurred, a set of key regulatory events leading to the G1/S boundary, including the expression of cyclins D2, E, A1, and B1, interleukin (IL)-2, IL-4, interferon-γ (IFN-γ), and activating protein-1 (AP-1), was examined. Cpd 6A suppressed, in activated T lymphocytes, the production and mRNA expression of cyclin E, IL-2, IL-4, IL-10, and IFN-γ in a dose-dependent manner. Expression of AP-1 proteins, consisting of c-Fos and c-Jun, in activated T lymphocytes was decreased by Cpd 6A. The kinetic study indicated that the inhibitory effects of Cpd 6A on IL-2 mRNA expressed in T cells might be related to blocking c-Fos protein synthesis. T-cell proliferation after Cpd 6A treatment was partially restored by addition of IL-2, IL-4, and IFN-γ. These suppressant effects of Cpd 6A on T-cell proliferation, activated by PHA, appeared to be mediated, at least in part, through the inhibition of early gene transcripts, especially those of cyclin E, IFN-γ, IL-2, and IL-4, and by arresting cell cycle progression in the cells. PMID:14504132

  17. Intercorrelation between immunological biomarkers and job stress indicators among female nurses: A nine-month longitudinal study

    Directory of Open Access Journals (Sweden)

    Hyung-Suk eYoon

    2014-10-01

    Full Text Available Some immunological biomarkers have been reported to be associated with job stress. This study was conducted to explore an intercorrelation between the psychological components of job stress and various immunological biomarkers among female nurses. To assess monthly and weekly job stress, 41 nurses had repeatedly completed the questionnaires such as the GJSQ, the POMS and the CES-D. Using flow cytometry and radioimmunoassay, the number of white blood cells, lymphocytic proliferation to mitogens, and toxoid were measured. Moreover, the levels of hydrocortisol, IL-b, INF-r, and TNF-a and salivary IgA were eveluated by enzyme-linked immunosorbent assay. When the Pearson correlation coefficients between job stress and immunological biomarkers were estimated after adjusting for age and smoking status, Clashes: conflict at work was significantly related to the number of CD4 cells (r = 0.36, p-value < 0.05, CD4 to CD8 ratio (0.35; < 0.05, response to concanavalin A (0.42; < 0.05, and phytohemagglutinin (0.35; < 0.05. Additionally, the level of hydrocortisol was significantly related to seven psychosocial measures; i.e., role conflict (-0.47; < 0.01, role ambiguity (-0.39; < 0.05, clashes at work (-0.38; < 0.05, control & influence at work (0.53; < 0.01, task control (0.55; < 0.001, resources at work (0.35; < 0.05, and skill underutilization (0.43; < 0.05. The results indicate that 1 the psychological job stress is associated with the levels of some immunological biomarkers in nurses; and 2 especially, hydrocortisol shows a remarkable relationship with diverse job stress indicators.

  18. Alpha-amylase inhibitor-1 gene from Phaseolus vulgaris expressed in Coffea arabica plants inhibits alpha-amylases from the coffee berry borer pest.

    Science.gov (United States)

    Barbosa, Aulus E A D; Albuquerque, Erika V S; Silva, Maria C M; Souza, Djair S L; Oliveira-Neto, Osmundo B; Valencia, Arnubio; Rocha, Thales L; Grossi-de-Sa, Maria F

    2010-06-17

    Coffee is an important crop and is crucial to the economy of many developing countries, generating around US$70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei), is responsible for worldwide annual losses of around US$500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an alpha-amylase inhibitor gene (alpha-AI1), which confers resistance against the coffee berry borer insect-pest, into C. arabica plants. We transformed C. arabica with the alpha-amylase inhibitor-1 gene (alpha-AI1) from the common bean, Phaseolus vulgaris, under control of the seed-specific phytohemagglutinin promoter (PHA-L). The presence of the alpha-AI1 gene in six regenerated transgenic T1 coffee plants was identified by PCR and Southern blotting. Immunoblotting and ELISA experiments using antibodies against alpha-AI1 inhibitor showed a maximum alpha-AI1 concentration of 0.29% in crude seed extracts. Inhibitory in vitro assays of the alpha-AI1 protein against H. hampei alpha-amylases in transgenic seed extracts showed up to 88% inhibition of enzyme activity. This is the first report showing the production of transgenic coffee plants with the biotechnological potential to control the coffee berry borer, the most important insect-pest of crop coffee.

  19. Comparison of the micronucleus and chromosome aberration techniques for the documentation of cytogenetic damage in radiochemotherapy-treated patients with rectal cancer

    Energy Technology Data Exchange (ETDEWEB)

    Wolff, Henrik Andreas; Hennies, Steffen; Herrmann, Markus Karl Alfred [Goettingen Univ. Medicine (DE). Dept. of Radiotherapy and Radiooncology] (and others)

    2011-01-15

    Purpose: The goal of the interdisciplinary Clinical Research Unit KFO179 (Biological Basis of Individual Tumor Response in Patients with Rectal Cancer) is to develop an individual Response and Toxicity Score for patients with locally advanced rectal cancer treated with neoadjuvant radiochemotherapy. The aim of the present study was to find a reliable and sensitive method with easy scoring criteria and high numbers of cell counts in a short period of time in order to analyze DNA damage in peripheral blood lymphocytes. Thus, the cytokinesis-block micronucleus (CBMN) assay and the chromosome aberration technique (CAT) were tested. Materials and Methods: Peripheral blood lymphocytes obtained from 22 patients with rectal cancer before (0 Gy), during (21.6 Gy), and after (50.4 Gy) radiochemotherapy were stimulated in vitro by phytohemagglutinin (PHA); the cultures were then processed for the CBMN assay and the CAT to compare the two methods. Results: A significant increase of chromosomal damage was observed in the course of radiochemotherapy parallel to increasing radiation doses, but independent of the chemotherapy applied. The equivalence of both methods was shown by Westlake's equivalence test. Conclusion: The results show that the CBMN assay and the CAT are equivalent. For further investigations, we prefer the CBMN assay, because it is simpler through easy scoring criteria, allows high numbers of cell counts in less time, is reliable, sensitive, and has higher statistical power. In the future, we plan to integrate cytogenetic damage during radiochemotherapy into the planned Response and Toxicity Score within our interdisciplinary Clinical Research Unit. (orig.)

  20. Assessment of genetic damage in peripheral blood of human volunteers exposed (whole-body) to a 200 muT, 60 Hz magnetic field.

    Science.gov (United States)

    Albert, Genevieve C; McNamee, James P; Marro, Leonora; Bellier, Pascale V; Prato, Frank S; Thomas, Alex W

    2009-02-01

    To investigate the extent of damage in nucleated cells in peripheral blood of healthy human volunteers exposed to a whole-body 60 Hz, 200 microT magnetic field. In this study, 10 male and 10 female healthy human volunteers received a 4 h whole-body exposure to a 200 microT, 60 Hz magnetic field. In addition, five males and five females were treated in a similar fashion, but were exposed to sham conditions. For each subject, a blood sample was obtained prior to the exposure period and aliquots were used as negative- (pre-exposure) and positive- [1.5 Gray (Gy) (60)Cobalt ((60)Co) gamma-irradiation] controls. At the end of the 4 h exposure period, a second blood sample was obtained. The extent of DNA damage was assessed in peripheral human blood leukocytes from all samples using the alkaline comet assay. To detect possible clastogenic effects, the incidence of micronuclei was assessed in phytohemagglutinin (PHA)-stimulated lymphocytes using the cytokinesis-block micronucleus assay. There was no evidence of either increased DNA damage, as indicated by the alkaline comet assay, or increased incidence of micronuclei (MN) in the magnetic field exposed group. However, an in vitro exposure of 1.5 Gy gamma-irradiation caused a significant increase in both DNA damage and MN induction. This study found no evidence that an acute, whole-body exposure to a 200 microT, 60 Hz magnetic field for 4 hours could cause DNA damage in human blood.

  1. DNA-AP sites generation by Etoposide in whole blood cells

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    Valverde Mahara

    2009-11-01

    Full Text Available Abstract Background Etoposide is currently one of the most commonly used antitumor drugs. The mechanisms of action proposed for its antitumor activity are based mainly on its interaction with topoisomerase II. Etoposide effects in transformed cells have been described previously. The aim of the present study was to evaluate the genotoxic effects of this drug in non-transformed whole blood cells, such as occurs as collateral damage induced by some chemotherapies. Methods To determine etoposide genotoxicity, we employed Comet assay in two alkaline versions. To evaluate single strand breaks and delay repair sites we use pH 12.3 conditions and pH >13 to evidence alkali labile sites. With the purpose to quantified apurinic or apyrimidine (AP sites we employed a specific restriction enzyme. Etoposide effects were determined on whole blood cells cultured in absence or presence of phytohemagglutinin (PHA treated during 2 and 24 hours of cultured. Results Alkaline (pH > 13 single cell gel electrophoresis (SCGE assay experiments revealed etoposide-induced increases in DNA damage in phytohemaglutinine (PHA-stimulated blood and non-stimulated blood cells. When the assay was performed at a less alkaline pH, 12.3, we observed DNA damage in PHA-stimulated blood cells consistent with the existence of alkali labile sites (ALSs. In an effort to elucidate the molecular events underlying this result, we applied exonuclease III (Exo III in conjunction with a SCGE assay, enabling detection of DNA-AP sites along the genome. More DNA AP-sites were revealed by Exo III and ALSs were recognized by the SCGE assay only in the non-stimulated blood cells treated with etoposide. Conclusion Our results indicate that etoposide induces DNA damage specifically at DNA-AP sites in quiescent blood cells. This effect could be involved in the development of secondary malignancies associated with etoposide chemotherapy.

  2. Lymphotoxin prevention of diethylnitrosamine carcinogenesis in vivo.

    Science.gov (United States)

    Ransom, J H; Evans, C H; DiPaolo, J A

    1982-09-01

    Development of intervention measures to control cancer would be facilitated by being able to monitor in vivo carcinogenesis by in vitro quantitation of early indices of neoplastic transformation to assess the in vivo effectiveness of preventive-therapeutic measures. Pregnant Syrian golden hamsters were used in an in vivo-in vitro transplacental model of carcinogenesis to determine the extent that in vivo administration of immunologic hormone preparations along with chemical carcinogen would prevent morphologic transformation assessed in vitro. Pregnant hamsters at 10-11 days of gestation were given injections ip of 3 mg diethylnitrosamine (DENA)/100 g body weight and were killed 2 days later when fetal cells were seeded for colony formation. The frequency of morphologically transformed colonies was assessed after 7 days of growth. Cloning efficiency and mean transformation frequency after DENA exposure were 3.6% and 1 X 10(-4) per cell seeded, respectively. The ip injection of an immunologic hormone preparation reduced the transformation frequency by 46%. The hormone preparation, containing 10,000 U of lymphotoxin but no detectable interferon, was the ultrafiltered lymphokines (greater than 10,000 mol wt) from phytohemagglutinin-stimulated hamster peritoneal leukocytes. The effect of lymphotoxin on cocarcinogenic exposure of fetal cells to DENA in vivo followed by X-irradiation in vitro was also determined. Cells exposed to 250 rad in vitro had a cloning efficiency of 0.5% and a transformation frequency of 0.4 X 10(-4) per cell seeded. After DENA injection and X-irradiation, the transformation frequency increased to 1 X 10(-4) and was inhibited 64% by lymphotoxin in vivo. Thus immunologic hormones (e.g., lymphotoxin) can prevent carcinogenesis in vivo. Furthermore, in vitro quantitation of transformation is a rapid means for evaluating therapeutic and autochthonous effector mechanisms for their ability to prevent or otherwise modulate carcinogenesis in vivo.

  3. Lymphocyte transformation suppression caused by pyoderma--failure to demonstrate it in uncomplicated demodectic mange.

    Science.gov (United States)

    Barta, O; Waltman, C; Oyekan, P P; McGrath, R K; Hribernik, T N

    1983-01-01

    Three dogs with demodectic mange uncomplicated by a bacterial infection and 9 dogs with demodectic mange and pyoderma were tested for their lymphocyte response to phytomitogens in vitro and for the presence of the serum's lymphocyte immunoregulatory factors (SLIF) suppressing blastogenesis. None of the 3 dogs with uncomplicated demodectic mange showed any detectable dysfunction of their lymphocytes or presence of the blastogenesis suppressing SLIF. Their lymphocytes generally responded to the mitogens with more blastogenesis than lymphocytes from healthy controls. On the other hand, in the group of 9 dogs with demodicosis complicated by a bacterial infection, high levels of the blastogenesis suppressing SLIF for concanavalin A-sensitive cells were detected in 4 dogs, for phytohemagglutinin-sensitive cells in 2 dogs, and for pokeweed mitogen-sensitive cells in 1 (of only 3 tested) dog. Dysfunction of lymphocytes per se (detected by a decreased blastogenesis in nonsuppressive normal canine and bovine sera) was detected in 3 dogs with demodicosis with pyoderma. The success of the treatment of demodectic mange or the bacterial skin infection did not correlate with the previous presence or absence of the blastogenesis suppressing SLIF. The treatment of pyoderma was less successful in dogs with an increase in blastogenesis of unstimulated cells in fresh normal canine serum over that in autologous serum. All 3 dogs with a detected dysfunction of their lymphocytes either died or were euthanatized as untreatable cases. It is concluded that the development of demodectic mange per se did not cause the appearance of the blastogenesis suppressing SLIF, which was primarily related to the appearance and extent of the secondary bacterial skin infection.

  4. Expression of protooncogenes during lymphocyte activation by growth factors.

    Science.gov (United States)

    Bulanova, E G; Budagyan, V M; Yarilin, A A; Mazurenko, N N

    1997-09-01

    Effects of growth factors of non-immune origin including somatotropin (ST) and platelet-derived growth factor (PDGF) on the expression of the proteins encoded by c-fos, c-myc, c-fun, and c-ets family protooncogenes were studied for the first time. The dynamics of the oncoprotein expression in activated CD(3+)-lymphocytes was investigated by immunoblotting. The accumulation of the Fos and Myc proteins was enhanced in T-lymphocytes treated with ST, PDGF, or phytohemagglutinin; the accumulation was maximum at 30-60 min and decreased in 2 h; the data indicate that the oncoproteins participate in the early lymphocyte activation by various growth factors. The Jun protein appears only in 3 h after the onset of lymphocyte activation; this suggests independent participation of Fos in the early stages of lymphocyte activation prior to the appearance of Jun, preceding the joint action of Fos and Jun within the AP-1 transcription complex. The products of the c-ets family are differentially activated by the studied growth factors. Resting lymphocytes actively accumulate the Ets-1 protein; ST and PDGF activation decreases Ets-1 expression in 2 h. The Ets-2 protein is not detected in resting cells and PDGF-activated lymphocytes, whereas lymphocyte activation by ST is associated with accumulation of Ets-2. The data suggest that the product of the c-ets-1 gene is more important in the regulation of resting cells and the product of the c-ets-2 gene is important during activation of lymphocytes by ST. The results indicate that activation of lymphocytes with growth factors of non-immune origin is mediated by several signal transduction pathways.

  5. Evaluation of immune responses in HIV infected patients with pleural tuberculosis by the QuantiFERON® TB-Gold interferon-gamma assay

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    Lekabe Jacob M

    2008-03-01

    Full Text Available Abstract Background Diagnosis of tuberculous (TB pleuritis is difficult and better diagnostic tools are needed. New blood based interferon-gamma (IFN-γ tests are promising, but sensitivity could be low in HIV positive patients. The IFN-γ tests have not yet been validated for use in pleural fluid, a compartment with higher level of immune activation than in blood. Methods The QuantiFERON TB®-Gold (QFT-TB test was analysed in blood and pleural fluid from 34 patients presenting with clinically suspected pleural TB. Clinical data, HIV status and CD4 cell counts were recorded. Adenosine deaminase activity (ADA analysis and TB culture were performed on pleural fluid. Results The patients were categorised as 'confirmed TB' (n = 12, 'probable TB' (n = 16 and 'non-TB' pleuritis (n = 6 based on TB culture results and clinical and biochemical criteria. The majority of the TB patients were HIV infected (82%. The QFT-TB in pleural fluid was positive in 27% and 56% of the 'confirmed TB' and 'probable TB' cases, respectively, whereas the corresponding sensitivities in blood were 58% and 83%. Indeterminate results in blood (25% were caused by low phytohemagglutinin (PHA = positive control IFN-γ responses, significantly lower in the TB patients as compared to the 'non-TB' cases (p = 0.02. Blood PHA responses correlated with CD4 cell count (r = 0.600, p = 0.028. In contrast, in pleural fluid indeterminate results (52% were caused by high Nil (negative control IFN-γ responses in both TB groups. Still, the Nil IFN-γ responses were lower than the TB antigen responses (p Conclusion The QFT-TB test in blood could contribute to the diagnosis of TB pleuritis in the HIV positive population. Still, the number of inconclusive results is too high to recommend the commercial QFT-TB test for routine use in pleural fluid in a TB/HIV endemic resource-limited setting.

  6. Autoradiographic detection of HPRT variants of human lymphocytes resistant to RNA synthesis inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Jones, I.M.; Zetterberg, G.; Strout, C.L.; Carrano, A.V.

    1985-01-01

    The feasibility of using RNA synthesis in freshly isolated, human peripheral blood lymphocytes to detect 6-thioguanine (TG)- and 8-azaguanine (AG)-resistant variants in an autoradiographic assay similar to that of Strauss and Albertini (1979) has been evaluated. In phytohemagglutinin (PHA)-stimulated cultures RNA synthesis and HPRT activity began well in advance of DNA synthesis and increased in parallel during the first 44 h of culture. Introduction of TG or AG with PHA at the beginning of culture completely inhibited DNA synthesis during the first 44 h and reduced RNA synthesis to low levels within 24 h. When TG or AG was added after cells had been in culture for 38 h, DNA synthesis was reduced quickly while RNA synthesis was inhibited more slowly. An autoradiographic assay is described in which freshly isolated lymphocytes are cultured with PHA for 24 h, with or without TG or AG, then labeled with (/sup 3/H)uridine for 1 h. TG-resistant and AG-resistant variant frequencies for 2 normal individuals and a Lesch-Nyhan individual were determined with this assay. The variant frequencies for the normal individuals ranged from 0.46 to 10.6 x 10/sup -5/ depending upon the selective conditions used. All the Lesch-Nyhan cells were resistant to 0.2 ..mu..M-2 mM AG; some were sensitive to 0.2 mM TG and most were sensitive to 2.0 mM TG. 24 references, 3 figures, 1 table.

  7. Discodermolide--a new, marine-derived immunosuppressive compound. I. In vitro studies.

    Science.gov (United States)

    Longley, R E; Caddigan, D; Harmody, D; Gunasekera, M; Gunasekera, S P

    1991-10-01

    The in vitro immunosuppressive properties of a novel, marine-derived compound, discodermolide, are reported here. Discodermolide suppressed the proliferative responses of splenocytes in the murine two-way mixed lymphocyte reaction (MLR) and concanavalin A stimulated cultures, with IC50 values of 0.24 microM and 0.19 microM, respectively. There was no evidence of cytotoxicity for murine splenocytes at concentrations of discodermolide as high as 1.26 microM. Similarly, discodermolide suppressed the proliferative responses of human peripheral blood leukocytes (PBL) in the two-way MLR, and Con A and phytohemagglutinin mitogenesis. The IC50 values were 5.65 microM, 28.02 microM, and 30.12 microM for the MLR, Con A, and PHA mitogenic responses, respectively. There was no evidence of cytotoxicity toward human PBL at discodermolide concentrations as high as 80.64 microM. Discodermolide was equally effective, compared with cyclosporine, in suppressing the PMA-ionomycin induced proliferation of purified, murine T cells, with IC50 values of 9.0 nM and 14.0 nM for discodermolide and CsA, respectively. The production of IL-2 by PMA-ionomycin stimulated T cells was not inhibited by discodermolide; however, the percentage of IL-2 receptor-bearing cells as measured by immunofluorescence with 7D4 antibody, specific for the 55-kDa chain (p55) comprising the murine IL-2 receptor, was reduced. The expression of a similar chain comprising the human IL-2 receptor (Tac antigen, p55) by PHA or Con-A-stimulated PBL was similarly suppressed by discodermolide. The precise mechanism of action of discodermolide remains to be elucidated.

  8. Differential effects of early- and late-life access to carotenoids on adult immune function and ornamentation in mallard ducks (Anas platyrhynchos.

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    Michael W Butler

    Full Text Available Environmental conditions early in life can affect an organism's phenotype at adulthood, which may be tuned to perform optimally in conditions that mimic those experienced during development (Environmental Matching hypothesis, or may be generally superior when conditions during development were of higher quality (Silver Spoon hypothesis. Here, we tested these hypotheses by examining how diet during development interacted with diet during adulthood to affect adult sexually selected ornamentation and immune function in male mallard ducks (Anas platyrhynchos. Mallards have yellow, carotenoid-pigmented beaks that are used in mate choice, and the degree of beak coloration has been linked to adult immune function. Using a 2 × 2 factorial experimental design, we reared mallards on diets containing either low or high levels of carotenoids (nutrients that cannot be synthesized de novo throughout the period of growth, and then provided adults with one of these two diets while simultaneously quantifying beak coloration and response to a variety of immune challenges. We found that both developmental and adult carotenoid supplementation increased circulating carotenoid levels during dietary treatment, but that birds that received low-carotenoid diets during development maintained relatively higher circulating carotenoid levels during an adult immune challenge. Individuals that received low levels of carotenoids during development had larger phytohemagglutinin (PHA-induced cutaneous immune responses at adulthood; however, dietary treatment during development and adulthood did not affect antibody response to a novel antigen, nitric oxide production, natural antibody levels, hemolytic capacity of the plasma, or beak coloration. However, beak coloration prior to immune challenges positively predicted PHA response, and strong PHA responses were correlated with losses in carotenoid-pigmented coloration. In sum, we did not find consistent support for either the

  9. Cycling and Tai Chi Chuan exercises exert greater immunomodulatory effect on surface antigen expression of human hepatitis B virus

    Institute of Scientific and Technical Information of China (English)

    CHEN Yu-yawn; CHIANG Jasson; CHEN Yu-jen; CHEN Kung-tung; YANG Rong-sen; LIN Jaung-geng

    2008-01-01

    Background Both athletes with intensive exercise and aged people may have weakened immunity against virus infection.This study aimed to evaluate whether people undergoing aerobic exercises including competitive cyctists with moderate training (CMT) and middle-aged people practicing Tai Chi Chuan (TCC) exercise have higher immunity against hepatitis B virus than age-matched sedentary controls including college students (CSC) and middle-aged people (MSC).Methods Human peripheral blood mononuclear cells from competitive cyclists and sedentary controls were stimulated by phytohemagglutinin (PHA) to prepare conditioned medium (MNC-CM) for the assessment of inhibitory effects on hepatitis B surface antigen (HBsAg) expression in human hepatoma Hep3B cells.Results The inhibitory effects on the relative HBsAg expression of CMT's and TCC's MNC-CM were greater than those of the controls.The CMT's MNC-CM prepared from 5 pg/ml PHA decreased HBsAg expression to 61.5%,whereas that of CSC remained at 83.8%.Similarly,this expression by treatment of TCC group' MNC-CM was 68.4% whereas that of MSC group was 84.3%.The levels of cytokines such as interferon-y (IFN-y),tumor necrosis factor-a (TNF-α),IFN-α and interleukin-1β(1L-1β) in the MNC-CM from the CMT and TCC groups were greater than those in the controls.Antibody neutralization of CMT's MNC-CM and addition of recombinant cytokines into CSC's MNC-CM indicated that IFN-y,TNF-α and IFN-α had synergistic effects against HBsAg expression.Similar blocking effect was noted in TCC versus MSC groups.Conclusion These results suggest that the immunomodulatory response to suppress HBsAg expression in CMT and TCC with moderate aerobic exercise is greater than that in age-matched sedentary controls.

  10. Abnormally High Expression of BAFF on T Lymphocytes from Lung Cancer-associated Pleural Effusions and Its Potent Anti-tumor Effect

    Institute of Scientific and Technical Information of China (English)

    Haiyan XU; Xiaozhou HE; Yibei ZHU; Tiangzan YAN; Hongbin MA; Xueguang ZHANG

    2007-01-01

    In the present study, the expressions of B cell activating factor belonging to the tumor necrosis factor family (BAFF) and its receptors (BAFF-R and TACI) on T lymphocytes from malignant pleural effusion (MPE) were examined by fluorescence-activated cell sorting (FACS) analysis, and compared with those on the T lymphocytes from non-malignant pleural effusion (NMPE) and healthy controls. It was found that CD3 positive T lymphocytes (including CD4, CD8, and part of CD25 and CD69 positive cells) of MPE in lung cancer highly and consistently expressed the BAFF molecule, while high expressions of BAFF could only be found in phytohemagglutinin (PHA) or interleukin 2 (IL-2) induced T lymphocytes from NMPE or healthy controls. These results were consistent with the results from BAFF mRNA detection by real-time PCR. In addition, T lymphocytes from MPE expressed significantly more BAFF-R than those from NMPE or healthy controls, while the expression of TACI was increased on CD4+ T cells but decreased on CD8+ T cells when compared with controls. The Annexin/PI assay suggested that recombinant human BAFF (rhBAFF) could promote the survival rate of T lymphocytes from MPE, while the decoy receptor TACI-Fc fusion protein could promote the apoptosis rate of T lymphocytes. Cytokines in the supernatant detected by ELISA assay showed that rhBAFF could significantly upregulate the secretion of IFN-γ in vitro,and the IFN-γ level in the TACI-Fc-treated group resembled that of the control groups. All of these results indicated that the abnormally high expression of BAFF on T lymphocytes from MPE may play a role of antitumor effect.

  11. Inhibitory Effects of Berberine on the Activation and Cell Cycle Progression of Human Peripheral Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    Lihui Xu; Yi Liu; Xianhui He

    2005-01-01

    The immunosuppressive property of berberine, an isoquinoline alkaloid, has been well documented, but the mechanism of its action on lymphocytes has not been completely elucidated. The present study is to investigate the effect of berberine on the activation and proliferation of lymphocytes, in particular T lymphocytes. Whole peripheral blood from healthy donors was stimulated with phytohemagglutinin (PHA) alone or phorbol dibutyrate (PDB) plus ionomycin, and the expression of CD69 and CD25 on T lymphocytes was evaluated with flow cytometry.The distribution of cell cycles and cell viability were analyzed by staining with propidium iodide (PI) and 7-aminoactinomycin D (7-AAD), respectively. The results showed that 100 μmol/L and 50 μmol/L of berberine significantly inhibited CD69 expression on T cells stimulated with PDB plus ionomycin or PHA, whereas the effect of 25 μmol/L berberine was not significant. As the incubation time increased, the extent of inhibition decreased.Similarly, the expression of CD25 was also reduced by berberine in a dose-dependent manner over the concentration range of 25-100 μmol/L. Besides, this alkaloid could block lymphocyte cell cycle progression from G0/G1 phase to S and G2/M phase without phase specificity. Moreover, analysis following 7-AAD staining revealed that berberine had no significant cytotoxicity on lymphocytes. Taken together, berberine significantly inhibits the expression of activation antigens on T lymphocytes and also blocks the progression of cell cycles of lymphocytes,suggesting that berberine may exert immunosuppressive effect through inhibiting the activation and proliferation of T cells.

  12. Cocoa procyanidins and human cytokine transcription and secretion.

    Science.gov (United States)

    Mao, T; Van De Water, J; Keen, C L; Schmitz, H H; Gershwin, M E

    2000-08-01

    We examined whether cocoa, in its isolated procyanidin fractions (monomer through decamer), would modulate cytokine production at the levels of transcription and protein secretion in both resting and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC). In resting cells, interleukin (IL)-1beta and IL-4 gene expression from cocoa-treated cells varied markedly among the subjects tested. However, at the protein level, the larger fractions (pentamer through decamer) stimulated a dramatic increase in IL-1beta concentration (up to ninefold) with increasing degree of polymerization. Similarly, these larger fractions augmented IL-4 concentration by as much as 2 pg/ml, whereas the control displayed levels nearly undetectable. In the presence of PHA, gene expression also seemed to be most affected by the larger procyanidin fractions. The pentameric through decameric fractions increased IL-1 beta expression by 7-19% compared with PHA control, whereas the hexameric through decameric fractions significantly inhibited PHA-induced IL-4 transcription in the range of 71-86%. This observation at the transcription level for IL-1 beta was reflected at the protein level in PHA-stimulated PBMC. Significant reductions in mitogen-induced IL-4 production were also seen at the protein level with the hexamer, heptamer and octamer. Individual oligomeric cocoa fractions were unstimulatory for IL-2 in resting PBMC. However, when induced with PHA, the pentamer, hexamer and heptamer fractions caused a 61-73% inhibition in IL-2 gene expression. This study offers additional data for the consideration of the health benefits of dietary polyphenols from a wide variety of foods, including those benefits associated specifically with cocoa and chocolate consumption.

  13. Effects of environmental stressors on lymphocyte proliferation in Florida manatees, Trichechus manatus latirostris.

    Science.gov (United States)

    Walsh, Cathy J; Luer, Carl A; Noyes, David R

    2005-02-10

    The health of many Florida manatees (Trichechus manatus latirostris) is adversely affected each year by exposure to cold weather or harmful algal blooms (red tide; Karenia brevis). Exposures can be sublethal, resulting in stressed animals that are rescued and taken to authorized facilities for rehabilitation, or lethal if exposures are prolonged or unusually severe. To investigate whether sublethal environmental exposures can impair immune function in manatees, rendering animals vulnerable to disease or death, mitogen-induced proliferation was assessed in lymphocytes from manatees exposed to cold temperatures (N=20) or red tide (N=19) in the wild, and compared to lymphocyte responses from healthy free-ranging manatees (N=32). All animals sampled for this study were adults. Lymphocytes were stimulated in vitro with either concanavalin A (ConA) or phytohemagglutinin (PHA) and proliferation was assessed after 96 h using incorporation of the thymidine analog, bromodeoxyuridine (BrdU), into newly synthesized DNA. Proliferation of lymphocytes from manatees rescued from exposure to red tide or cold-stress was approximately one-third that of lymphocytes from healthy free-ranging manatees. To examine the direct effects of red tide toxins on lymphocyte function, mitogen-induced proliferation was assessed following co-culture of lymphocytes with K. brevis toxin extracts. Stimulation indices decreased with increasing toxin concentration, with a significant decrease in proliferation occurring in the presence of 400 ng red tide toxins/ml. When lymphocytes from cold-stressed manatees were co-cultured with red tide toxin extracts, proliferative responses were reduced even further, suggesting multiple stressors may have synergistic effects on immune function in manatees.

  14. 半滑舌鳎外周血淋巴细胞培养条件研究%Study on blood lymphocytic cell cultivation of half smooth tongue sole ( Cynoglossus semilaevis)

    Institute of Scientific and Technical Information of China (English)

    王美玉; 刘海金; 范兆廷; 宋立民

    2012-01-01

    以半滑舌鳎为试验对象,对淋巴细胞体外培养和染色体制备条件如促分裂剂、培养时间、培养温度、培养基、秋水仙素浓度及作用时间等进行探索,在建立一套基于半滑舌鳎淋巴细胞体外培养的染色体制备方法.结果表明,对于半滑舌鳎淋巴细胞体外培养,用离心法收集淋巴细胞,培养基为含15%体积分数胎牛血清的L-15培养基,添加3μg·mL-1的PHA-P作促分裂剂,分裂指数可达2.6%±0.2%,6d后收集细胞制作染色体,获得较好效果.%Half smooth tongue sole(Cynogtossus semilaevis) was used to explore the condition of the peripheral blood lymphocytic cell cultivation and chromosome preparation. The conditions were searched from mrtogens, incubation periods, culture temperature, media and the effect of the chloride. The aim was to establish a systemic method. The results showed that lymphocytes isolated from fresh blood by a stirring method were cultured in medium L-15 supplemented with 15%FBS, 3 μg·mL-1 PHA (phytohemagglutinin) as mitogens, and collected at day 6. This was the optimized culture condition for half smooth tongue sole. After culture to make chromosome, the mitotic index (Ml) could reach (2.6±0.2)%.

  15. Long-term fungal inhibitory activity of water-soluble extracts of Phaseolus vulgaris cv. Pinto and sourdough lactic acid bacteria during bread storage.

    Science.gov (United States)

    Coda, Rossana; Rizzello, Carlo G; Nigro, Franco; De Angelis, Maria; Arnault, Philip; Gobbetti, Marco

    2008-12-01

    The antifungal activity of proteinaceous compounds from different food matrices was investigated. In initial experiments, water-soluble extracts of wheat sourdoughs, cheeses, and vegetables were screened by agar diffusion assays with Penicillium roqueforti DPPMAF1 as the indicator fungus. Water-soluble extracts of sourdough fermented with Lactobacillus brevis AM7 and Phaseolus vulgaris cv. Pinto were selected for further study. The crude water-soluble extracts of L. brevis AM7 sourdough and P. vulgaris cv. Pinto had a MIC of 40 mg of peptide/ml and 30.9 mg of protein/ml, respectively. MICs were markedly lower when chemically synthesized peptides or partially purified protein fractions were used. The water-soluble extract of P. vulgaris cv. Pinto showed inhibition toward a large number of fungal species isolated from bakeries. Phaseolin alpha-type precursor, phaseolin, and erythroagglutinating phytohemagglutinin precursor were identified in the water-soluble extract of P. vulgaris cv. Pinto by nano liquid chromatography-electrospray ionization-tandem mass spectrometry. When the antifungal activity was assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, all three proteins were inhibitory. A mixture of eight peptides was identified from the water-soluble extract of sourdough L. brevis AM7, and five of these exhibited inhibitory activity. Bread was made at the pilot plant scale by sourdough fermentation with L. brevis AM7 and addition of the water-soluble extract (27%, vol/wt; 5 mg of protein/ml) of P. vulgaris cv. Pinto. Slices of bread packed in polyethylene bags did not show contamination by fungi until at least 21 days of storage at room temperature, a level of protection comparable to that afforded by 0.3% (wt/wt) calcium propionate.

  16. Implementation of exon arrays: alternative splicing during T-cell proliferation as determined by whole genome analysis

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    Whistler Toni

    2010-09-01

    Full Text Available Abstract Background The contribution of alternative splicing and isoform expression to cellular response is emerging as an area of considerable interest, and the newly developed exon arrays allow for systematic study of these processes. We use this pilot study to report on the feasibility of exon array implementation looking to replace the 3' in vitro transcription expression arrays in our laboratory. One of the most widely studied models of cellular response is T-cell activation from exogenous stimulation. Microarray studies have contributed to our understanding of key pathways activated during T-cell stimulation. We use this system to examine whole genome transcription and alternate exon usage events that are regulated during lymphocyte proliferation in an attempt to evaluate the exon arrays. Results Peripheral blood mononuclear cells form healthy donors were activated using phytohemagglutinin, IL2 and ionomycin and harvested at 5 points over a 7 day period. Flow cytometry measured cell cycle events and the Affymetrix exon array platform was used to identify the gene expression and alternate exon usage changes. Gene expression changes were noted in a total of 2105 transcripts, and alternate exon usage identified in 472 transcript clusters. There was an overlap of 263 transcripts which showed both differential expression and alternate exon usage over time. Gene ontology enrichment analysis showed a broader range of biological changes in biological processes for the differentially expressed genes, which include cell cycle, cell division, cell proliferation, chromosome segregation, cell death, component organization and biogenesis and metabolic process ontologies. The alternate exon usage ontological enrichments are in metabolism and component organization and biogenesis. We focus on alternate exon usage changes in the transcripts of the spliceosome complex. The real-time PCR validation rates were 86% for transcript expression and 71% for

  17. Clinical grade expansion of CD45RA, CD45RO, and CD62L-positive T-cell lines from HLA-compatible donors: high cytotoxic potential against AML and ALL cells.

    Science.gov (United States)

    Barbui, Anna M; Borleri, Gianmaria; Conti, Elena; Ciocca, Alice; Salvi, Anna; Micò, Caterina; Introna, Martino; Rambaldi, Alessandro

    2006-04-01

    Identification of a clinical grade method for the ex vivo generation of donor-derived T cells cytotoxic against both myeloid and lymphoblastic cells still remains elusive. We investigated rapid generation and expansion of donor derived-allogeneic T-cell lines cytotoxic against patient leukemic cells. Acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL) blasts were cultured 5 days in Stem Span, granulocyte macrophage colony-stimulating factor, interleukin-4, and calcium ionophore. All B-precursor ALL (N22) and AML (N13), but not T-cell ALL (N3), differentiated into mature leukemia-derived antigen-presenting cells (LD-APC). All but one LD-APC generated cytotoxic T lymphocyte (CTL) from adult human leukocyte antigen (HLA)-identical (N8) or unrelated donors (N2). Upon in vitro culture, donor-derived CTL acquired a memory T phenotype, showing concomitant high CD45RA, CD45RO, CD62L expression. CD8(+) cells, but not CD4(+) cells, were granzyme, perforine, and interferon-gamma-positive. Pooled CD4(+) and CD8(+) cells were cytotoxic against leukemic blasts (32%, 30:1 E:T ratio), but not against autologous or patient-derived phytohemagglutinin blasts. LD-APC from five ALL patients were used to generate CTL from cord blood. A mixed population of CD4(+) and CD8(+) cells was documented in 54% of wells. T cells acquired classical effector memory phenotype and showed a higher cytotoxicity against leukemia blasts (47%, 1:1 E:T ratio). Adult and cord blood CTL showed a skewing from a complete T-cell receptor repertoire to an oligo-clonal/clonal pattern. Availability of these cells should allow clinical trials for salvage treatment of leukemia patients relapsing after allogeneic stem cell transplantation.

  18. The clinical implications of mixed lymphocyte reaction with leukemic cells.

    Science.gov (United States)

    Kim, Hee-Je; Kim, Tai-Gyu; Cho, Hyun Il; Han, Hoon; Min, Woo-Sung; Kim, Chun-Choo

    2002-11-01

    To evaluate the clinical implications of a mixed lymphocyte reaction between leukemic cells and lymphocytes from HLA-matched sibling donors, we attempted to generate donor-derived, graft-versus-leukemia-effective cells and to define their characteristics. We studied 8 patients with chronic myelogenous leukemia (CML), including 5 patients in the chronic phase (CP), 3 patients in the accelerated phase (AP), and 2 patients with acute myelogenous leukemia (AML) in their first complete remission. Cells from these patients were used as stimulators in a mixed lymphocyte reaction.The effects of natural killer (NK) cells and cytotoxic T-lymphocytes (CTLs) were separated by observing tests for cytotoxicity to target cells, including K562 cells, the patient's leukemic cells, and phytohemagglutinin (PHA) blasts. Donor-derived antileukemic CTLs againstthe patient's own leukemic cells are productive in vitro. The efficacy of generating CTLs against leukemic target cells was (in decreasing order) AML, CML-CP, and CML-AP. Cytotoxic activity against leukemic targets was prominent in 4 cases--2 CML-CP and the 2 AML cases. On the contrary, the 3 cases of CML-AP showed low CTL activity. In cases showing 1 positive result among 3 targets (K562 cells, the patient's leukemic cells, and PHA blasts), the relapse rate was significantly lower (P = .022) on follow-up (median, 33 months; 7-40 months) after hematopoietic stem cell transplantation. By a combined analysis of the cytotoxicity effects for all 3 target cells, we were able to demonstrate a correlation between leukemic relapse and the variable degree of the cytotoxicity test results. Although the total sample numbers for this study were low, we speculate that these results may come from differences in the individual characteristics of the leukemic cells that are in line with their clinical disease status.

  19. The aberrant asynchronous replication — characterizing lymphocytes of cancer patients — is erased following stem cell transplantation

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    Korenstein-Ilan Avital

    2010-05-01

    Full Text Available Abstract Background Aberrations of allelic replication timing are epigenetic markers observed in peripheral blood cells of cancer patients. The aberrant markers are non-cancer-type-specific and are accompanied by increased levels of sporadic aneuploidy. The study aimed at following the epigenetic markers and aneuploidy levels in cells of patients with haematological malignancies from diagnosis to full remission, as achieved by allogeneic stem cell transplantation (alloSCT. Methods TP53 (a tumor suppressor gene assigned to chromosome 17, AML1 (a gene assigned to chromosome 21 and involved in the leukaemia-abundant 8;21 translocation and the pericentomeric satellite sequence of chromosome 17 (CEN17 were used for replication timing assessments. Aneuploidy was monitored by enumerating the copy numbers of chromosomes 17 and 21. Replication timing and aneuploidy were detected cytogenetically using fluorescence in situ hybridization (FISH technology applied to phytohemagglutinin (PHA-stimulated lymphocytes. Results We show that aberrant epigenetic markers are detected in patients with hematological malignancies from the time of diagnosis through to when they are scheduled to undergo alloSCT. These aberrations are unaffected by the clinical status of the disease and are displayed both during accelerated stages as well as in remission. Yet, these markers are eradicated completely following stem cell transplantation. In contrast, the increased levels of aneuploidy (irreversible genetic alterations displayed in blood lymphocytes at various stages of disease are not eliminated following transplantation. However, they do not elevate and remain unchanged (stable state. A demethylating anti-cancer drug, 5-azacytidine, applied in vitro to lymphocytes of patients prior to transplantation mimics the effect of transplantation: the epigenetic aberrations disappear while aneuploidy stays unchanged. Conclusions The reversible nature of the replication aberrations may

  20. Induction of adaptive response in human blood lymphocytes exposed to radiofrequency radiation.

    Science.gov (United States)

    Sannino, Anna; Sarti, Maurizio; Reddy, Siddharth B; Prihoda, Thomas J; Vijayalaxmi; Scarfì, Maria Rosaria

    2009-06-01

    The incidence of micronuclei was evaluated to assess the induction of an adaptive response to non-ionizing radiofrequency (RF) radiation in peripheral blood lymphocytes collected from five different human volunteers. After stimulation with phytohemagglutinin for 24 h, the cells were exposed to an adaptive dose of 900 MHz RF radiation used for mobile communications (at a peak specific absorption rate of 10 W/kg) for 20 h and then challenged with a single genotoxic dose of mitomycin C (100 ng/ml) at 48 h. Lymphocytes were collected at 72 h to examine the frequency of micronuclei in cytokinesis-blocked binucleated cells. Cells collected from four donors exhibited the induction of adaptive response (i.e., responders). Lymphocytes that were pre-exposed to 900 MHz RF radiation had a significantly decreased incidence of micronuclei induced by the challenge dose of mitomycin C compared to those that were not pre-exposed to 900 MHz RF radiation. These preliminary results suggested that the adaptive response can be induced in cells exposed to non-ionizing radiation. A similar phenomenon has been reported in cells as well as in animals exposed to ionizing radiation in several earlier studies. However, induction of adaptive response was not observed in the remaining donor (i.e., non-responder). The incidence of micronuclei induced by the challenge dose of mitomycin C was not significantly different between the cells that were pre-exposed and unexposed to 900 MHz RF radiation. Thus the overall data indicated the existence of heterogeneity in the induction of an adaptive response between individuals exposed to RF radiation and showed that the less time-consuming micronucleus assay can be used to determine whether an individual is a responder or non-responder.

  1. Molybdate modulates mitogen and cyclosporin responses of human peripheral blood lymphocytes.

    Science.gov (United States)

    Michelis, Fotios V; Delitheos, Andreas; Tiligada, Ekaterini

    2011-07-01

    The trace element molybdenum (Mo) is an essential component of key physiological systems in animals, plants and microorganisms. The molybdate oxoanion MoO(4)(2-) has been demonstrated to cause diverse yet poorly understood biochemical and pharmacological effects, such as non-specific inhibition of phosphatases and stabilization of steroid receptors. This study aimed to investigate the effects of molybdate on the activation of human peripheral blood lymphocytes (hPBLs) ex vivo and its potential interaction with the widely used immunosuppressant drug cyclosporin A (CsA). Lymphocyte activation was evaluated by performing multiple experiments determining blastogenesis in cultured peripheral blood lymphocytes obtained from 5 healthy volunteers, following stimulation induced by phytohemagglutinin (PHA), in the absence or presence of 0.05-10 mM sodium molybdate or/and 2.5-30 μg/mL CsA. Blastogenesis was assessed by a morphometric assay based on the relative proportions of unactivated lymphocytes, activated lymphoblasts and cells with aberrant morphology after PHA-induced activation. Molybdate concentrations up to 1 mM showed no effect on lymphocyte blastogenesis, while higher concentrations exerted immunosuppressive actions on cultured hPBLs. Co-administration of 0.1 mM sodium molybdate with CsA, at doses up to 20 μg/mL, induced no alteration in the response of cultured hPBLs to CsA. However, molybdate potentiated the immunosuppressive action of higher CsA concentrations, implying a likely dose-related synergistic interaction of the two agents in PHA-stimulated blood lymphocytes. These observations are indicative of the possible biological importance of molybdate oxoanions in the modulation of hPBL activation that may have pharmacological consequences during the therapeutic application of immunomodulatory drugs.

  2. Cytotoxic activities of Euphorbia kopetdaghi against OVCAR-3 and EJ-138 cell lines

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    Aghaei Mahmoud

    2015-04-01

    Full Text Available Introduction: Over the centuries, the genus Euphorbia was known to be toxic to humans and animals. Recently, in a primary study significant suppressive activity against phytohemagglutinin activated T-cell proliferation has been reported from this plant. Therefore, this study was designed to evaluate the cytotoxic effects of different parts of E. kopetdaghi against cancer cell lines. Methods: Filtration and in vacuo concentration resulted in a green gum which was subjected on silica gel CC (hexane/Acetone, 0→50 to several fractions: F1-F8. The inhibitory effects of obtained fractions with 5, 50, and 500 μg/ml concentrations were evaluated on proliferation and viability of cancer cells (OVCAR and EJ-138 in 48 hours treatment. Finally, cell viability was determined at a wavelength of 570 by 3-4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT method. Results: Based on studies of microscopic observation and viability testing, F1, F2, F4, F5, F6, and F7 showed significant cytotoxic effect at concentration of 50 and 500 μg/ml against EJ-138 and OVCAR-3 cell lines. These fractions inhibited growth of EJ-138 and OVCAR-3 cells in a concentration-dependent manner. Fraction of F8 induced tumor promotion significantly in EJ-138 and OVCAR-3 cells, respectively. Conclusion: Due to the inhibitory properties of E. kopetdaghi extract and its fractions on cancer cells of OVCAR3 and EJ-13, isolation, purification and identification of compounds presented in the fractions possessing cytotoxic effects are recommended which were the area of our future research.

  3. Impact of Dietary Protein Concentration and Quality on Immune Function of Cats

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    Paßlack, Nadine; Kohn, Barbara; Doherr, Marcus G.; Zentek, Jürgen

    2017-01-01

    Protein levels and quality in cat food can vary significantly and might affect immune function in various ways. In the present study, 3 diets with a low protein quality (LQ) and 3 diets with a high protein quality (HQ) were offered to 10 healthy adult cats for 6 weeks each, using a randomized cross-over design. The LQ and HQ diets differed in the collagen content and had low (36.7% and 36.2%), medium (45.0% and 43.3%) and high (56.1% and 54.9%) protein levels. At the end of each feeding period, blood was collected for phenotyping of leukocyte subsets, lymphocyte proliferation assay and cytokine measurements, phagocytosis assay and differential blood count. The results demonstrated no group differences for numbers of CD4+CD8-, CD4+CD8+, CD4-CD8+, MHCII+, CD21+, SWC3+ and CD14+ cells in the blood of the cats. Proliferative activity of lymphocytes when stimulated with pokeweed mitogen, Concanavalin A and Phytohemagglutinin, M form did not differ depending on the dietary protein concentration and quality. Concentrations of tumor necrosis factor alpha and interferon gamma in the supernatant of the proliferation assay were also not affected by the dietary treatment. Blood monocyte phagocytic activity was higher (P = 0.048) and cell numbers of eosinophilic granulocytes in the blood were lower (P = 0.047) when cats were fed the low protein diets. In conclusion, only a few differences in feline immune cell populations and activity depending on dietary protein supply could be detected. However, the observed increase of eosinophilic granulocytes by a higher protein intake indicates an activation of immunological mechanisms and requires further investigation. PMID:28072882

  4. Ratite oils promote keratinocyte cell growth and inhibit leukocyte activation

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    Bennett, Darin C.; Leung, Gigi; Wang, Eddy; Ma, Sam; Lo, Blanche K. K.; McElwee, Kevin J.; Cheng, Kimberly M.

    2015-01-01

    Traditionally, native Australian aborigines have used emu oil for the treatment of inflammation and to accelerate wound healing. Studies on mice suggest that topically applied emu oil may have anti-inflammatory properties and may promote wound healing. We investigated the effects of ratite oils (6 emu, 3 ostrich, 1 rhea) on immortalized human keratinocytes (HaCaT cells) in vitro by culturing the cells in media with oil concentrations of 0%, 0.5%, and 1.0%. Peking duck, tea tree, and olive oils were used as comparative controls. The same oils at 0.5% concentration were evaluated for their influence on peripheral blood mononuclear cell (PBMC) survival over 48 hr and their ability to inhibit IFNγ production in PBMCs activated by phytohemagglutinin (PHA) in ELISpot assays. Compared to no oil control, significantly shorter population doubling time durations were observed for HaCaT cells cultured in emu oil (1.51 × faster), ostrich oil (1.46 × faster), and rhea oil (1.64 × faster). Tea tree oil demonstrated significant antiproliferative activity and olive oil significantly prolonged (1.35 × slower) cell population doubling time. In contrast, almost all oils, particularly tea tree oil, significantly reduced PBMC viability. Different oils had different levels of inhibitory effect on IFNγ production with individual emu, ostrich, rhea, and duck oil samples conferring full inhibition. This preliminary investigation suggests that emu oil might promote wound healing by accelerating the growth rate of keratinocytes. Combined with anti-inflammatory properties, ratite oil may serve as a useful component in bandages and ointments for the treatment of wounds and inflammatory skin conditions. PMID:26217022

  5. Transport of proteins to the plant vacuole is not by bulk flow through the secretory system, and requires positive sorting information.

    Science.gov (United States)

    Dorel, C; Voelker, T A; Herman, E M; Chrispeels, M J

    1989-02-01

    Plant cells, like other eukaryotic cells, use the secretory pathway to target proteins to the vacuolar/lysosomal compartment and to the extracellular space. We wished to determine whether the presence of a hydrophobic signal peptide would result in the transport of a reporter protein to vacuoles by bulk flow; to investigate this question, we expressed a chimeric gene in transgenic tobacco. The chimeric gene, Phalb, used for this study consists of the 1,188-bp 5' upstream sequence and the hydrophobic signal sequence of a vacuolar seed protein phytohemagglutinin, and the coding sequence of a cytosolic seed albumin (PA2). The chimeric protein PHALB cross-reacted with antibodies to PA2 and was found in the seeds of the transgenic plants (approximately 0.7% of total protein), but not in the leaves, roots, or flowers. Immunoblot analyses of seed extracts revealed four glycosylated polypeptides ranging in molecular weight from 29,000 to 32,000. The four polypeptides are glycoforms of a single polypeptide of Mr 27,000, and the heterogeneity is due to the presence of high mannose and endoglycosidase H-resistant glycans. The PHALB products reacted with an antiserum specific for complex plant glycans indicating that the glycans had been modified in the Golgi apparatus. Subcellular fractionation of glycerol extracts of mature seeds showed that only small amounts of PHALB accumulated in the protein storage vacuoles of the tobacco seeds. In homogenates made in an isotonic medium, very little PHALB was associated with the organelle fraction containing the endoplasmic reticulum and Golgi apparatus; most of it was in the soluble fraction. We conclude that PHALB passed through the Golgi apparatus, but did not arrive in the vacuoles. Transport to vacuoles is not by a bulk-flow mechanism, once proteins have entered the secretory system, and requires information beyond that provided by a hydrophobic signal peptide.

  6. Telomerase Activity in Peripheral Blood Mononuclear Cells from Senile Patients with Pneumonia

    Institute of Scientific and Technical Information of China (English)

    LIU Jian; ZHOU Zhen; LIU Xiaoqing

    2006-01-01

    To investigate the changes of the activity of telomerase in peripheral blood mononuclear cells (PBMCs) from senile patients with pneumonia, the telomerase activity was examined before and after the stimulation of phytohemagglutinin-M (PHA-M) in PBMCs from 10 control subjects (group A), 12 non-senile patients with pneumonia (group B) and 9 senile patients with pneumonia (group C). Also observed was the proliferative response of these PBMCs to PHA-M. The results showed that, both with or without the stimulation of PHA-M, the values of telomerase activity in PBMCs from group C patients (A values: pre-stimulation, 0.43±0.04; post-stimulation, 0.63±0.03) were significantly lower than those in PBMCs from both group A patients (A values: prestimulation, 0.65±0.05;post-stimulation, 1.26±0.13;P<0.001, respectively) and group B patients (A values: pre-stimulation, 0.63±0.03; post-stimulation, 0.93±0.03;P<0.05, respectively). The results of MTT test showed that the proliferative activity of PBMCs in group C patients (A value: 0.35±0.03) was also significantly lower than that in group A patients (A value:0. 55±0.04; P<0.05) and group B patients (A value: 0.46±0.03;P<0.05). These results indicate that the telomerase activity decreases in senile patients with pneumonia, which may be one of the mechanisms for the weakened immune function in those patients.

  7. Nutritional quality of legumes, and their role in cardiometabolic risk prevention: a review.

    Science.gov (United States)

    Bouchenak, Malika; Lamri-Senhadji, Myriem

    2013-03-01

    Legumes (including alfalfa, clover, lupins, green beans and peas, peanuts, soybeans, dry beans, broad beans, dry peas, chickpeas, and lentils) represent an important component of the human diet in several areas of the world, especially in the developing countries, where they complement the lack of proteins from cereals, roots, and tubers. In some regions of the world, legume seeds are the only protein supply in the diet. The health benefits of legume consumption have received rising interest from researchers, and their consumption and production extends worldwide. Among European countries, higher legume consumption is observed around the Mediterranean, with per capita daily consumption between 8 and 23 g, while in Northern Europe, the daily consumption is less than 5 g per capita. The physiological effects of different legumes vary significantly. These differences may result from the polysaccharides composition, in particular, the quantity and variety of dietary fibers and starch, protein make-up, and variability in phytochemical content. The majority of legumes contain phytochemicals: bioactive compounds, including enzyme inhibitors, phytohemagglutinins (lectins), phytoestrogens, oligosaccharides, saponins, and phenolic compounds, which play metabolic roles in humans who frequently consume these foods. Dietary intake of phytochemicals may provide health benefits, protecting against numerous diseases or disorders, such as coronary heart disease, diabetes, high blood pressure and inflammation. The synergistic or antagonistic effects of these phytochemical mixtures from food legumes, their interaction with other components of the diet, and the mechanism of their action have remained a challenge with regard to understanding the role of phytochemicals in health and diseases. Their mitigating effects and the mechanism of their action need to be further addressed if we are to understand the role of phytochemicals in health and diseases. This review provides an overview

  8. TUMOR NECROSIS FACTOR-ALPHA POLYMORPHISM AND SECRETION IN MYASTHENIA GRAVIS

    Institute of Scientific and Technical Information of China (English)

    Yu-zhou Guan; Li-ying Cui; Yan-feng Li; Jun-bao Zhang

    2005-01-01

    Objective To analyze the relationship between tumor necrosis factor-alpha (TNFo) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFo in peripheral blood mononuclear cells (PBMC) in MG patients.Methods A biallelic polymorphism at position -308 in the promoter of TNFα gene was screened by PCR amplification and NcoI recognition site. One hundred and twenty-three MG cases and 115 healthy controls were included in this study. MG patients were classified to different groups according to clinical type, age at onset, and sex respectively. PBMC were isolated from 20 patients and 20 healthy controls, and then cultured in the presence or absence of phytohemagglutinin (PHA) and acetycholine receptors (AchR). The supernatants were harvested after incubation and stored until TNFαwas assayed by enzyme-linked immunosorbent assay.Results The frequency of TNFα-308 allele 2 (A) was found significantly increase in MG patients and showed a trend especially in late onset (≥ 40 years) and male patients (P < 0.05). The allele A had no relationship with thymic pathogenesis in MG patients. But frequency of allele A was significantly higher in general type than in ocular type (P < 0.05). MG patients had a higher inducible level of TNFα by PHA and AchR, and could be down regulated after treatment.Conclusion Polymorphism in TNFα gene promoter -308 is associated with onset of MG. The microsatellite allele TNFα2 confer risk for the development of MG in Chinese patients. MG patients have a higher inducible level of TNFα.

  9. Human T-lymphotropic virus type 1-infected cells secrete exosomes that contain Tax protein.

    Science.gov (United States)

    Jaworski, Elizabeth; Narayanan, Aarthi; Van Duyne, Rachel; Shabbeer-Meyering, Shabana; Iordanskiy, Sergey; Saifuddin, Mohammed; Das, Ravi; Afonso, Philippe V; Sampey, Gavin C; Chung, Myung; Popratiloff, Anastas; Shrestha, Bindesh; Sehgal, Mohit; Jain, Pooja; Vertes, Akos; Mahieux, Renaud; Kashanchi, Fatah

    2014-08-08

    Human T-lymphotropic virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia and HTLV-1-associated myelopathy/tropical spastic paraparesis. The HTLV-1 transactivator protein Tax controls many critical cellular pathways, including host cell DNA damage response mechanisms, cell cycle progression, and apoptosis. Extracellular vesicles called exosomes play critical roles during pathogenic viral infections as delivery vehicles for host and viral components, including proteins, mRNA, and microRNA. We hypothesized that exosomes derived from HTLV-1-infected cells contain unique host and viral proteins that may contribute to HTLV-1-induced pathogenesis. We found exosomes derived from infected cells to contain Tax protein and proinflammatory mediators as well as viral mRNA transcripts, including Tax, HBZ, and Env. Furthermore, we observed that exosomes released from HTLV-1-infected Tax-expressing cells contributed to enhanced survival of exosome-recipient cells when treated with Fas antibody. This survival was cFLIP-dependent, with Tax showing induction of NF-κB in exosome-recipient cells. Finally, IL-2-dependent CTLL-2 cells that received Tax-containing exosomes were protected from apoptosis through activation of AKT. Similar experiments with primary cultures showed protection and survival of peripheral blood mononuclear cells even in the absence of phytohemagglutinin/IL-2. Surviving cells contained more phosphorylated Rb, consistent with the role of Tax in regulation of the cell cycle. Collectively, these results suggest that exosomes may play an important role in extracellular delivery of functional HTLV-1 proteins and mRNA to recipient cells.

  10. Toxic effects of dietary methylmercury on immune system development in nestling American kestrels (Falco sparverius)

    Science.gov (United States)

    Fallacara, Dawn M.; Halbrook, Richard S.; French, John B.

    2011-01-01

    This study evaluated the effects of dietary methylmercury (MeHg) on immune system development in captive-reared nestling American kestrels (Falco sparverius) to determine whether T cell–mediated and antibody-mediated adaptive immunity are targets for MeHg toxicity at environmentally relevant concentrations. Nestlings received various diets, including 0 (control), 0.6, and 3.9 μg/g (dry wt) MeHg for up to 18 d posthatch. Immunotoxicity endpoints included cell-mediated immunity (CMI) using the phytohemagglutinin (PHA) skin-swelling assay and antibody-mediated immune response via the sheep red blood cell (SRBC) hemagglutination assay. T cell– and B cell–dependent histological parameters in the spleen, thymus, and bursa of Fabricius were correlated with the functional assays. For nestlings in the 0.6 and 3.9 μg/g MeHg groups, CMI was suppressed by 73 and 62%, respectively, at 11 d of age. Results of this functional assay were correlated with T cell–dependent components of the spleen and thymus. Dose-dependent lymphoid depletion in spleen tissue directly affected the proliferation of T-lymphocyte populations, insofar as lower stimulation indexes from the PHA assay occurred in nestlings with lower proportions of splenic white pulp and higher THg concentrations. Nestlings in the 3.9 μg/g group also exhibited lymphoid depletion and a lack of macrophage activity in the thymus. Methylmercury did not have a noticeable effect on antibody-mediated immune function or B cell–dependent histological correlates. We conclude that T cell–mediated immunosuppression is the primary target of MeHg toward adaptive immunity in developing kestrels. This study provides evidence that environmentally relevant concentrations of MeHg may compromise immunocompetence in a developing terrestrial predator and raises concern regarding the long-term health effects of kestrels that were exposed to dietary MeHg during early avian development.

  11. Novel compound heterozygous mutations in ZAP70 in a Chinese patient with leaky severe combined immunodeficiency disorder.

    Science.gov (United States)

    Liu, Qing; Wang, Yan-Ping; Liu, Qiao; Zhao, Qin; Chen, Xue-Mei; Xue, Xiu-Hong; Zhou, Li-Na; Ding, Yuan; Tang, Xue-Mei; Zhao, Xiao-Dong; Zhang, Zhi-Yong

    2017-01-26

    In humans, the complete lack of tyrosine kinase ZAP70 function results in combined immunodeficiency (CID), with abnormal thymic development and defective T cell receptor (TCR) signaling of peripheral T cells, characterized by the selective absence of CD8(+) T cells. So far, 15 unique ZAP70 mutations have been identified in approximately 20 patients with CID, with variable clinical presentations. Herein, we report the first case from China of novel compound heterozygous mutations in ZAP70 (c.598-599delCT, p.L200fsX28; c.847 C>T, R283H). The patient suffered from early-onset and recurrent infections, but showed normal growth and development without signs of failure to thrive, thus presenting as leaky SCID. The patient also had clinical manifestations of autoimmunity, such as eczematous skin lesion, inflammatory bowel disease (IBD), and intractable diarrhea, suggesting compromised T cell tolerogenic functions. Residual ZAP70 expression was identified. Immunological analysis revealed the selective absence of CD8(+) T cells in the periphery and the presence of CD4(+) T cells that failed to respond to phytohemagglutinin. Stimulation with lectin from pokeweed mitogen also failed to stimulate B cell proliferation in the patient. The frequency of Tfhs and Tregs in the patient was lower compared with the normal reference. Compared with the age-matched healthy control, the level of IL-17 was higher and the levels of IFN-γ, IL-4, and IL-21 were lower. Infants with selected CD8 deficiency and severe autoimmune disorders or exaggerated inflammation should be screened for ZAP70 deficiency.

  12. Effects of sustained sleep restriction on mitogen-stimulated cytokines, chemokines and T helper 1/ T helper 2 balance in humans.

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    John Axelsson

    Full Text Available BACKGROUND: Recent studies suggest that acute sleep deprivation disrupts cellular immune responses by shifting T helper (Th cell activity towards a Th2 cytokine profile. Since little is known about more long-term effects, we investigated how five days of sleep restriction would affect pro-inflammatory, chemotactic, Th1- and Th2 cytokine secretion. METHODS: Nine healthy males participated in an experimental sleep protocol with two baseline sleep-wake cycles (sleep 23.00-07.00 h followed by 5 days with restricted sleep (03.00-07.00 h. On the second baseline day and on the fifth day with restricted sleep, samples were drawn every third hour for determination of cytokines/chemokines (tumor necrosis factor alpha (TNF-α, interleukin (IL -1β, IL-2, IL-4 and monocyte chemoattractant protein-1 (MCP-1 after in vitro stimulation of whole blood samples with the mitogen phytohemagglutinin (PHA. Also leukocyte numbers, mononuclear cells and cortisol were analysed. RESULTS: 5-days of sleep restriction affected PHA-induced immune responses in several ways. There was a general decrease of IL-2 production (p<.05. A shift in Th1/Th2 cytokine balance was also evident, as determined by a decrease in IL2/IL4 ratio. No other main effects of restricted sleep were shown. Two significant interactions showed that restricted sleep resulted in increased TNF-α and MCP-1 in the late evening and early night hours (p's<.05. In addition, all variables varied across the 24 h day. CONCLUSIONS: 5-days of sleep restriction is characterized by a shift towards Th2 activity (i.e. lower 1L-2/IL-4 ratio which is similar to the effects of acute sleep deprivation and psychological stress. This may have implications for people suffering from conditions characterized by excessive Th2 activity like in allergic disease, such as asthma, for whom restricted sleep could have negative consequences.

  13. Concentrations of cyclosporin A and FK506 that inhibit IL-2 induction in human T cells do not affect TGF-beta1 biosynthesis, whereas higher doses of cyclosporin A trigger apoptosis and release of preformed TGF-beta1.

    Science.gov (United States)

    Minguillón, Jordi; Morancho, Beatriz; Kim, Seong-Jin; López-Botet, Miguel; Aramburu, José

    2005-05-01

    Cyclosporin A (CsA) and FK506 suppress T cell activation by inhibiting calcineurin and the calcineurin-dependent transcription factors nuclear factor of activated T cells (NFATc), which are central regulators of T cell function. It was reported that CsA up-regulated the transcription of transforming growth factor-beta1 (TGF-beta1) in lymphocytes and other cells and activated its promoter in A549 lung carcinoma cells, but the mechanisms involved are poorly understood, and it is unclear whether calcineurin plays any role. We have studied the regulation of TGF-beta1 in normal human lymphocytes and cell lines. In Jurkat T cells, the TGF-beta1 promoter was activated by calcineurin and NFATc and inhibited by CsA and FK506. However, the promoter was insensitive to both drugs in A549 cells. In human T cells preactivated with phytohemagglutinin, biosynthesis of TGF-beta1, induced by the T cell receptor (TCR) or the TGF-beta receptor, was not substantially affected by CsA and FK506 concentrations (< or = 1 microM) that effectively inhibited interleukin-2 production. However, pretreatment of fresh lymphocytes with CsA or FK506 during primary TCR stimulation reduced their production of TGF-beta1 during secondary TCR activation. Finally, high concentrations of CsA (10 microM), in the range attained in vivo in experiments in rodents, caused apoptosis in human T cells and the release of preformed, bioactive TGF-beta1. These effects are unlikely to owe to calcineurin inhibition, as they were not observed with FK506. Our results indicate that CsA and FK506 are not general inducers of TGF-beta1 biosynthesis but can cause different effects on TGF-beta1 depending on the cell type and concentrations used.

  14. 21 Days head-down bed rest induces weakening of cell-mediated immunity - Some spaceflight findings confirmed in a ground-based analog.

    Science.gov (United States)

    Kelsen, Jens; Bartels, Lars Erik; Dige, Anders; Hvas, Christian Lodberg; Frings-Meuthen, Petra; Boehme, Gisela; Thomsen, Marianne Kragh; Fenger-Grøn, Morten; Dahlerup, Jens Frederik

    2012-08-01

    Several studies indicate a weakening of cell-mediated immunity (CMI) and reactivation of latent herpes viruses during spaceflight. We tested the hypothesis that head-down bed rest (HDBR), a ground-based analog of spaceflight, mimics the impact of microgravity on human immunity. Seven healthy young males underwent two periods of 3 weeks HDBR in the test facility of the German Aerospace Center. As a nutritional countermeasure aimed against bone demineralisation, 90 mmol potassium bicarbonate (KHCO(3)) was administered daily in a crossover design. Blood samples were drawn on five occasions. Whole blood was stimulated with antigen i.e. Candida albicans, purified protein derivative (PPD) tuberculin, tetanus toxoid and Cytomegalovirus (CMV) (CMV-QuantiFERON). Flow cytometric analysis included CD4(+)CD25(+)CD127(-)FOXP3(+) regulatory T cells (Tregs), γδ T cells, B cells, NK cells and dendritic cells. In one of the two bed rest periods, we observed a significant decrease in production of interleukin-2 (IL-2), interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) following phytohemagglutinin (PHA) stimulation, with a rapid normalization being observed after HDBR. The cytokine levels showed a V-shaped pattern that led to a relativeTh2-shift in cytokine balance. Only three individuals responded to the specific T cell antigens without showing signs of an altered response during HDBR, nor did we observe reactivation of CMV or Epstein-Barr virus (EBV). Of unknown significance, dietary supplementation with KHCO(3) counteracted the decrease in IL-2 levels during HDBR, while there was no impact on other immunological parameters. We conclude that discrete alterations in CMI may be induced by HDBR in selected individuals.

  15. Immunological and clinical observations in diabetic kidney graft recipients pretreated with total-lymphoid irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Waer, M.; Vanrenterghem, Y.; Roels, L.; Ang, K.K.; Bouillon, R.; Lerut, T.; Gruwez, J.; van der Schueren, E.; Vandeputte, M.; Michielsen, P.

    1987-03-01

    In a feasibility study, twenty patients with end-stage diabetic nephropathy were treated with fractionated total-lymphoid irradiation (TLI, mean dose 25 Gy), before transplantation of a first cadaveric kidney. During radiotherapy, only one patient had a serious side effect (bone marrow depression). After transplantation four patients died (one of a myocardial infarction, one of ketoacidosis, and two of infections occurring during treatment of rejection crises). One graft was lost because of chronic rejection. The other 15 patients have a functioning graft (mean follow-up 24 months) and receive low-dose prednisone alone (less than 10 mg/day, n = 11) or in conjunction with cyclosporine (n = 4) as maintenance immunosuppressive therapy. A favorable clinical outcome after TLI (no, or only one, steroid-sensitive rejection crisis) was significantly correlated with a high pre-TLI helper/suppressor lymphocyte ratio, a short interval between TLI and the time of transplantation, and the occurrence of functional suppressor cells early after TLI. The most striking immunological changes provoked by TLI consisted of a long-term depression of the mixed lymphocyte reaction and of the phytohemagglutinin, and Concanavalin A or pokeweed-mitogen-induced blastogenesis. A rapid and complete recovery of the natural killer cell activity was observed after TLI. A permanent inversion of the OKT4+ (T helper/inducer) over OKT8+ (T suppressor/cytotoxic) lymphocyte ratio was provoked by a decrease of the OTK4+ subpopulation, together with a supranormal recovery of the OKT8+ lymphocytes. A majority of the latter lymphocytes did also express the Leu 7 and the Leu 15 phenotype.

  16. Diphtheria toxin resistance in human lymphocytes and lymphoblasts in the in vivo somatic cell mutation test

    Energy Technology Data Exchange (ETDEWEB)

    Tomkins, D.J.; Wei, L.; Laurie, K.E.

    1985-01-01

    It has been shown that circulating peripheral blood lymphocytes can be used for the enumeration of 6-thioguanine-resistant cells that presumably arise by mutation in vivo. This somatic cell mutation test has been studied in lymphocytes from human populations exposed to known mutagens and/or carcinogens. The sensitivity of the test could be further enhanced by including other gene markers, since there is evidence for locus-specific differences in response to mutagens. Resistance to diphtheria toxin (Dip/sup r/) seemed like a potential marker to incorporate into the test because the mutation acts codominantly, can readily be selected in human diploid fibroblasts and Chinese hamster cells with no evidence for cell density or cross-feeding effects, and can be assayed for in nondividing cells by measuring protein synthesis inhibition. Blood samples were collected from seven individuals, and fresh, cryopreserved, or Epstein-Barr virus (EBV)-transformed lymphocytes were tested for continued DNA synthesis (TH-thymidine, autoradiography) or protein synthesis (TVS-methionine, scintillation counting). Both fresh and cryopreserved lymphocytes, stimulated to divide with phytohemagglutinin (PHA), continued to synthesize DNA in the presence of high doses of diphtheria toxin (DT). Similarly, both dividing (PHA-stimulated) and nondividing fresh lymphocytes carried on significant levels of protein synthesis even 68 hr after exposure to 100 flocculating units (LF)/ml DT. The results suggest that human T and B lymphocytes may not be as sensitive to DT protein synthesis inhibition as human fibroblast and Chinese hamster cells. For this reason, Dip/sup r/ may not be a suitable marker for the somatic cell mutation test.

  17. Monocyte chemoattractant protein-1 polymorphism interaction with spirulina immunomodulatory effects in healthy Korean elderly: A 16 week, double-blind randomized clinical trial

    Science.gov (United States)

    Park, Hee Jung

    2017-01-01

    BACKGROUND/OBJECTIVES Spirulina is a known a functional food related to lipid profiles, immune functions, and antioxidant capacity. Circulating monocyte chemoattractant protein-1 (MCP-1) level is associated with inflammation markers. Single nucleotide polymorphism in the MCP-1 promoter region -2518 have been identified and shown to affect gene transcription. Gene variation may also impact functional food supplementary effects. The current study investigated the interaction of MCP-1 -2518 polymorphism with spirulina supplements on anti-inflammatory capacity in Korean elderly. SUBJECTS/METHODS After genotyping, healthy elderly subjects (n = 78) were included in a randomized, double blind, and placebo controlled study. Baseline characteristic, body composition, and dietary intake were measured twice (baseline vs. week 16). For 16 weeks, subjects consumed 8 g either spirulina or placebo daily. Plasma MCP-1, interleukin (IL) -2, IL-6, tumor necrosis factor (TNF)-α, complement (C) 3, immunoglobulin (Ig) G, and Ig A concentrations and lymphocyte proliferation rate (LPR) were analyzed as inflammatory markers. RESULTS In the placebo group with A/A genotype, MCP-1 level was significantly increased, but the spirulina group with A/A genotype was unchanged. IL-2 was significantly increased only in subjects with spirulina supplementation. TNF-α was significantly reduced in subjects with the G carrier. C3 was significantly increased in the placebo group, particularly when A/A increased more than G, but not when spirulina was ingested. LPR was significantly different only in subjects with A/A genotype; there was a significant increase in phytohemagglutinin and lipopolysaccharide induced LPR in the spirulina group. CONCLUSION In healthy Korean elderly, spirulina supplementation may influence different inflammatory markers by the MCP-1 genotype. These results may be useful for customized dietary guidelines to improve immune function in Koreans. PMID:28765775

  18. The P2X7 loss-of-function Glu496Ala polymorphism affects ex vivo cytokine release and protects against the cytotoxic effects of high ATP-levels

    Directory of Open Access Journals (Sweden)

    Wesselius Anke

    2012-12-01

    Full Text Available Abstract Background The P2X7 receptor plays an important role in cytokine release during the inflammatory response in vivo. Polymorphisms within the P2X7 receptor gene that lead to loss of receptor function may contribute to impaired cytokine release by immune cells. Therefore, we investigated whether a known loss-of-function polymorphism (Glu496Ala in the P2X7 receptor gene leads to alterations in cytokine release in response to ATP. Results An ex vivo whole blood model was used to induce an inflammatory reaction with the pro-inflammatory stimuli LPS and PHA (phytohemagglutinin. Blood from n=9 subjects with the Glu496Ala P2X7 SNP (P2X7MUT and n=7 ‘wild-type’ subjects (no P2X7 SNP; P2X7WT was used. Addition of ATP (0.9-3 mM to LPS/PHA-stimulated whole blood induced an increase in IL-1β release in P2X7MUT subjects, whereas decreased release was observed in P2X7WT subjects. Decreased levels of IL-6 and TNF-α in response to ATP were shown in both P2X7MUT and P2X7WT subjects, which was less pronounced in P2X7MUT subjects. ATP at 3 mM also significantly decreased levels of lactate dehydrogenase (LDH in P2X7MUT subjects compared to P2X7WT subjects. Conclusions The presence of the non-synonymous Glu496Ala loss-of-function polymorphism within the P2X7 receptor gene is likely to be of importance in the release of cytokines during inflammation. Furthermore, this study suggests that carriers of the Glu496Ala loss-of-function polymorphism are protected against the cytotoxic effects of high ATP-levels.

  19. Peripheral blood mononuclear cell proliferation and cytokine production in sheep as affected by cortisol level and duration of stress.

    Science.gov (United States)

    Ciliberti, M G; Albenzio, M; Inghese, C; Santillo, A; Marino, R; Sevi, A; Caroprese, M

    2017-01-01

    A large number of studies recognize glucocorticoids (Gc) as suppressors of inflammation; Gc exert an important role in coordinating the magnitude and duration of host immune responses. In the present in vitro investigation, we tested incremental levels of cortisol to verify the immunosuppressive or immunopermissive role of cortisol in sheep peripheral blood mononuclear cells (PBMC) after acute and chronic stress. Phytohemagglutinin (PHA)-stimulated PBMC were cultured for 24h and 96h at 37°C with 5% of CO2 and varying cortisol levels: 10 ng/mL (baseline), 100 ng/mL (physiological poststressor), and 1,000 ng/mL [hyperactivated hypothalamic-pituitary-adrenal (HPA) axis]. The cell-free supernatants were collected for determination of IL-6, IL-1β, and IL-10 by ELISA, and the bromodeoxyuridine assay was performed on cells. Physiological cortisol concentration negatively affected the levels of IL-6 secreted by PBMC, resulting in increased cell proliferation after acute stress (24h of incubation). However, physiological cortisol concentration exhibited a reduction in cell proliferation induced by increased levels of IL-6 secreted by PBMC during chronic stress (96h of incubation). The cortisol concentration representing a hyperactivated HPA axis led to a reduction in cell proliferation after acute stress, which was probably induced by the elevated IL-10 production. Our results demonstrate that in sheep the effect of Gc on the immune system was related to the magnitude and the duration of stress. In particular, cortisol levels higher than physiological concentrations suppressed cell proliferation soon after acute stress. Instead, the physiological poststressor concentration of cortisol affected the immune responses in a bidirectional manner depending on the duration of the stressor. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  20. The PHA test reflects acquired T-cell mediated immunocompetence in birds.

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    José L Tella

    Full Text Available BACKGROUND: cological immunology requires techniques to reliably measure immunocompetence in wild vertebrates. The PHA-skin test, involving subcutaneous injection of a mitogen (phytohemagglutinin, PHA and measurement of subsequent swelling as a surrogate of T-cell mediated immunocompetence, has been the test of choice due to its practicality and ease of use in the field. However, mechanisms involved in local immunological and inflammatory processes provoked by PHA are poorly known, and its use and interpretation as an acquired immune response is currently debated. METHODOLOGY: Here, we present experimental work using a variety of parrot species, to ascertain whether PHA exposure produces larger secondary than primary responses as expected if the test reflects acquired immunocompetence. Moreover, we simultaneously quantified T-lymphocyte subsets (CD4(+, CD5(+ and CD8(+ and plasma proteins circulating in the bloodstream, potentially involved in the immunological and inflammatory processes, through flow cytometry and electrophoresis. PRINCIPAL FINDINGS: Our results showed stronger responses after a second PHA injection, independent of species, time elapsed and changes in body mass of birds between first and second injections, thus supporting the adaptive nature of this immune response. Furthermore, the concomitant changes in the plasma concentrations of T-lymphocyte subsets and globulins indicate a causal link between the activation of the T-cell mediated immune system and local tissue swelling. CONCLUSIONS/SIGNIFICANCE: These findings justify the widespread use of the PHA-skin test as a reliable evaluator of acquired T-cell mediated immunocompetence in diverse biological disciplines. Further experimental research should be aimed at evaluating the relative role of innate immunocompetence in wild conditions, where the access to dietary proteins varies more than in captivity, and to ascertain how PHA responses relate to particular host

  1. Arecoline inhibits interleukin-2 secretion in Jurkat cells by decreasing the expression of alpha7-nicotinic acetylcholine receptors and prostaglandin E2.

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    Hwang, G S; Hu, S; Lin, Y H; Chen, S T; Tang, T K; Wang, P S; Wang, S W

    2013-10-01

    The purpose of the present study was to explore the effect of arecoline on phytohemagglutinin (PHA)-stimulated interleukin-2 (IL-2) secretion, the expression of alpha7-nicotinic acetylcholine receptors (α7-nAChRs), prostaglandin E2(PGE2) protein, and IL-2 mRNA in human lymphocyte cells (Jurkat cell line). The IL-2 and PGE2 were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of phosphorylated extracellular signal-regulated kinase (ERK) and α7-nAChRs were determined by Western blotting. The level of IL-2 mRNA was determined by reverse-transcriptase polymerase chain reaction (RT-PCR). Arecoline, in a dose-dependent manner, significantly decreased IL-2 and PGE2 secretion by Jurkat cells incubated with 0 or 5 μg/ml 5 μg/ml PHA. PGE2 also significantly inhibited IL-2 secretion by Jurkat cells in a dose-dependent manner. In addition, reduced expression of PHA-induced ERK phosphorylation was observed in Jurkat cells treated with arecoline. PHA-enhanced IL-2 mRNA expression was also inhibited by arecoline. These results imply that arecoline inhibits the release of PGE2 and PHA-induced IL-2 secretion by Jurkat cells and that these effects seem to occur, at least in part, either through the attenuation of ERK in conjunction with a decrease of PHA-induced IL-2 mRNA expression. These results imply that arecoline inhibits the protein expression of α7-nAChRs , the release of PGE2 and PHA-induced IL-2 secretion by Jurkat cells.

  2. Effects of plant lectins on in vitro fibroblast proliferation

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    Ana Maria Sell

    2003-06-01

    Full Text Available Lectins are carbohydrate-binding proteins that have been isolated from various sources and presented a wide spectrum of biological activities. The effects of four lectins, namely, Phaseolus vulgaris phytohemagglutinin, PHA, wheat germ agglutinin, WGA, Artocarpus integrifolia seed lectins, jacalin and artocarpin, on in vitro fibroblasts proliferation were investigated. The lectins did not influence the initial cell adhesion to the plate. PHA and WGA at 10-20 µg/mL concentrations significantly decreased fibroblasts proliferation. At these concentrations, they caused morphological alterations on cells and over 80 µg/mL, promoted cell death. Neither jacalin nor artocarpin significantly affected cell proliferation.O objetivo deste trabalho foi avaliar a influência das lectinas PHA, WGA, jacalina e artocarpina sobre a proliferação de fibroblastos in vitro. Para tanto, fibroblastos gengivais de voluntários saudáveis foram cultivados, por cinco dias, em DMEM suplementado com soro bovino fetal (10% v/v e na presença das lectinas nas concentrações finais de 0.1 a 300 µg/mL. A adesão, o crescimento e a morfologia celular foram acompanhados por microscopia de inversão e contraste de fase. O índice de proliferação foi avaliado pelo método calorimétrico usando MTT. As lectinas não alteraram a adesão inicial dos fibroblastos à placa de poliestireno. PHA e WGA, nas concentrações de 10 a 20 µg/mL, diminuíram significativamente a proliferação celular. Nestas concentrações a morfologia celular é alterada e acima de 80 µg/mL, houve100% de morte celular. As lectinas jacalina e artocarpina não influenciaram a proliferação celular.

  3. Productive human immunodeficiency virus infection levels correlate with AIDS-related manifestations in the patient

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    Mathez, D.; Paul, D.; de Belilovsky, C.; Sultan, Y.; Deleuze, J.; Gorin, I.; Saurin, W.; Decker, R.; Leibowitch, J. (Univ. Rene-Descartes Paris-Ouest, Garches (France))

    1990-10-01

    Mononuclear cells were obtained from 71 human immunodeficiency virus type 1 (HIV-1) seropositive subjects presenting and first visit either as asymptomatic or with minor symptoms and with CD4 lymphocytes greater than 550 per mm3 (group A, 35 patients) or as patients with AIDS, AIDS-related illnesses, or CD4 lymphocytes less than 400 per mm3 (group B, 36 patients). After 1-5 years of follow-up, 13 patients of group A had essentially retained their initial status (asymptomatics); the 22 others had suffered clinical or immunological deterioration (progressors). Frozen cells were thawed and submitted to lethal gamma-irradiation in vitro (4500 rads; 1 rad = 0.01 Gy) before they were cultured with normal phytohemagglutinin-stimulated lymphocytes to determine radiation-resistant HIV expression ex vivo (R-HEV). HIV antigenemia correlated with R-HEV values in 142 samples (r = 0.92, P less than 0.001) but was a less sensitive predictor of disease than R-HEV. R-HEV was detected in all specimens from patients with major AIDS-related illnesses or HIV-associated CD4 lymphopenia. In 77% of the progressors from group A, R-HEV detection preceded the onset of AIDS-associated disease or CD4 lymphopenia by 1 year (average). Conversely, R-HEV was low or was not detected in 36 sequential specimens from the 13 patients who remained asymptomatic over the following 2-5 years. Thus, persistently low HIV expression in vivo predicted a nondiseased state, whereas higher HIV expression levels seemed necessary for disease to occur. These data indicate that R-HEV is related to productive HIV infection in vivo, the latter acting as a determinant of AIDS-related illnesses. In view of this, measurement of HIV expression levels in the patient should be useful in antiviral efficacy trials.

  4. Changes in some pro-and anti-inflammatory cytokines produced by bovine peripheral blood mononuclear cells following foot and mouth disease vaccination

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    N. Delirezh

    2016-09-01

    Full Text Available Interleukin (IL-17 is exclusively produced by CD4 helper T-cells upon activation. It most often acts as a pro-inflammatory cytokine, which stimulates the release of pro-inflammatory cytokines IL-6, IL-8, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF. In this study, we studied the in-vitro IL-17 response to specific antigens and a variety of mitogens and compared the IL-17 response to IL-2, IL-4, IL-5, IL-6, IL-10, and IFN-γ responses. We used a foot and mouth disease (FMD vaccine as specific antigens and mitogens (phytohemagglutinin [PHA], pokeweed mitogen [PWM], and concanavalin A [Con A] to stimulate peripheral blood mononuclear cells (PBMCs of vaccinated calves. Cell culture supernatant was harvested and analyzed for cytokines, using commercially available bovine ELISA kits. The mitogens induced a significant increase in IL-17 production. IL-17 was produced at high levels in response to the T cell-stimulated mitogens, PHA, and Con A, and at low levels in response to PWM mitogens. In contrast, level of the produced IL-17 cytokines in response to the FMDV antigens was lower as compared to those produced by mitogens. The FMDV antigens and mitogens significantly increased IL-17 production. There was not a correlation between IL-17 production and type-1 cytokine, IFN-γ, and IL-2, while there was a correlation between type-2 cytokine, IL-4, and IL-5 at either cytokine level produced by PBMCs stimulated by FMDV antigens. Moreover, there was an interaction between IL-17 and IL-6, that is, as IL-6 cytokine level elevated or diminished, IL-17 cytokine level increased or decreased, as well.

  5. Hydrophobic sodium fluoride-based nanocrystals doped with lanthanide ions: assessment of in vitro toxicity to human blood lymphocytes and phagocytes.

    Science.gov (United States)

    Sojka, Bartlomiej; Kuricova, Miroslava; Liskova, Aurelia; Bartusova, Maria; Banski, Mateusz; Misiewicz, Jan; Dusinska, Maria; Horvathova, Mira; Jahnova, Eva; Ilavska, Silvia; Szabova, Michaela; Rollerova, Eva; Podhorodecki, Artur; Tulinska, Jana

    2014-11-01

    In vitro immunotoxicity of hydrophobic sodium fluoride-based nanocrystals (NCs) doped with lanthanide ions was examined in this study. Although there is already a significant amount of optical and structural data on NaYF4 NCs, data on safety assessment are missing. Therefore, peripheral whole blood from human volunteers was used to evaluate the effect of 25 and 30 nm hydrophobic NaYF4 NCs dissolved in cyclohexane (CH) on lymphocytes, and of 10 nm NaYF4 NCs on phagocytes. In the concentration range 0.12-75 µg cm(-2) (0.17-106 µg ml(-1) ), both 25 and 30nm NaYF4 NCs did not induce cytotoxicity when measured as incorporation of [(3) H]-thymidine into DNA. Assessment of lymphocyte function showed significant suppression of the proliferative activity of T-lymphocytes and T-dependent B-cell response in peripheral blood cultures (n = 7) stimulated in vitro with mitogens phytohemagglutinin (PHA) and pokeweed (PWM) (PHA > PWM). No clear dose-response effect was observed. Phagocytic activity and respiratory burst of leukocytes (n = 5-8) were generally less affected. A dose-dependent suppression of phagocytic activity of granulocytes in cultures treated with 25 nm NCs was observed (vs. medium control). A decrease in phagocytic activity of monocytes was found in cells exposed to higher doses of 10 and 30 nm NCs. The respiratory burst of phagocytes was significantly decreased by exposure to the middle dose of 30 nm NCs only. In conclusion, our results demonstrate immunotoxic effects of hydrophobic NaYF4 NCs doped with lanthanide ions to lymphocytes and to lesser extent to phagocytes. Further research needs to be done, particularly faze transfer of hydrophobic NCs to hydrophilic ones, to eliminate the solvent effect.

  6. l-Arginine-Dependent Epigenetic Regulation of Interleukin-10, but Not Transforming Growth Factor-β, Production by Neonatal Regulatory T Lymphocytes

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    Kuender D. Yang

    2017-04-01

    Full Text Available A growing number of diseases in humans, including trauma, certain cancers, and infection, are known to be associated with l-arginine deficiency. In addition, l-arginine must be supplemented by diet during pregnancy to aid fetal development. In conditions of l-arginine depletion, T cell proliferation is impaired. We have previously shown that neonatal blood has lower l-arginine levels than adult blood, which is associated with poor neonatal lymphocyte proliferation, and that l-arginine enhances neonatal lymphocyte proliferation through an interleukin (IL-2-independent pathway. In this study, we have further investigated how exogenous l-arginine enhances neonatal regulatory T-cells (Tregs function in relation to IL-10 production under epigenetic regulation. Results showed that cord blood mononuclear cells (CBMCs produced higher levels of IL-10 than adult peripheral blood mononuclear cells (PBMCs by phytohemagglutinin stimulation but not by anti-CD3/anti-CD28 stimulation. Addition of exogenous l-arginine had no effect on transforming growth factor-β production by PBMCs or CBMCs, but enhanced IL-10 production by neonatal CD4+CD25+FoxP3+ Tregs. Further studies showed that IL-10 promoter DNA hypomethylation, rather than histone modification, corresponded to the l-arginine-induced increase in IL-10 production by neonatal CD4+ T cells. These results suggest that l-arginine modulates neonatal Tregs through the regulation of IL-10 promoter DNA methylation. l-arginine supplementation may correct the Treg function in newborns with l-arginine deficiency.

  7. Consequences of vitamin D receptor gene polymorphisms for growth inhibition of cultured human peripheral blood mononuclear cells by 1, 25-dihydroxyvitamin D3.

    Science.gov (United States)

    Colin, E M; Weel, A E; Uitterlinden, A G; Buurman, C J; Birkenhäger, J C; Pols, H A; van Leeuwen, J P

    2000-02-01

    In the vitamin D receptor (VDR) gene a BsmI restriction fragment length polymorphism (RFLP) in intron 8 and a translational start-site polymorphism, identified as a FokI RFLP, have been described. Crucial for a proper interpretation of these polymorphisms in association studies is the knowledge whether they have direct consequences for 1,25-(OH)2D3 action at cellular level. The present study was designed to assess functional significance of the FokI and BsmI VDR gene polymorphisms in peripheral blood mononuclear cells (PBMC) with a natural occurring VDR genotype for cell growth inhibition by 1,25-(OH)2D3. PBMC of women were isolated, VDR genotyped and in vitro inhibition by 1,25-(OH)2D3 of Phytohemagglutinin (PHA)-stimulated growth of PBMC was examined in relation to VDR genotype. PHA-stimulated growth and maximal growth inhibition were independent of VDR genotype. However, the FF genotype had a significant lower ED50 than the Ff genotype corresponding to an allele dose effect of 0.32 nM per f allele copy (P = 0.0036). For BsmI genotypes no differences in ED50 were observed. The present study demonstrates for the first time in cells with a natural VDR genotype a direct functional consequence of the VDR gene translational start-site polymorphism for the action of 1,25-(OH)2D3. Especially under conditions of vitamin D insufficiency these findings might have clinical implications.

  8. Toxicity Assessment of Common Beans (Phaseolus vulgaris L.) Widely Consumed by Tunisian Population.

    Science.gov (United States)

    Nciri, Nader; Cho, Namjun; El Mhamdi, Faiçal; Ben Ismail, Hanen; Ben Mansour, Abderraouf; Sassi, Fayçal Haj; Ben Aissa-Fennira, Fatma

    2015-09-01

    This research aimed at assessing the content and the functional properties of phytohemagglutinin (PHA) in different varieties of beans widely consumed in Tunisia through soaking, cooking, autoclaving, germination, and their combinations. This study was carried out on three varieties of white beans grown in different localities of Tunisia, namely Twila, Coco, and Beldia, as well as on imported and local canned beans. All bean samples underwent biochemical and immunological evaluation by employing several techniques such as indirect competitive enzyme-linked immunosorbent assay (ELISA), hemagglutinating assay, Ouchterlony double immunodiffusion, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Biochemical and immunological analyses indicated that raw dry beans contained a considerable amount of proteins and PHAs. ELISA demonstrated that soaking, either in plain water or in alkaline solution, caused an increase in the concentration of PHA. A slight increase of PHA was produced equally by germination during 4 days in all bean varieties. Cooking or autoclaving of presoaked beans resulted in a complete disappearance of PHA. ELISA test also proved that both imported and local canned beans contained fingerprints of PHA. Hemagglutination assays showed that not only cooked and autoclaved presoaked beans lacked the ability to agglutinate red blood cells but also autoclaved unsoaked beans did. In agar gel immunodiffusion using rabbit anti-PHA serum, raw, soaked, cooked unsoaked, and sprouted beans gave precipitin arc reactions, indicating that PHA existed in immunoreactive form in the tested seeds. SDS-PAGE electrophoretograms showed protein isolates of Twila and Beldia beans to have different profiles through soaking, cooking, and autoclaving processes. This work revealed that the combination of soaking and cooking/autoclaving was the best way in reducing PHA content and its activity in all bean varieties when compared with germination.

  9. Isolation of a T-cell clone showing HLA-DRB1*0405-restricted cytotoxicity for hematopoietic cells in a patient with aplastic anemia.

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    Nakao, S; Takami, A; Takamatsu, H; Zeng, W; Sugimori, N; Yamazaki, H; Miura, Y; Ueda, M; Shiobara, S; Yoshioka, T; Kaneshige, T; Yasukawa, M; Matsuda, T

    1997-05-15

    The existence of T cells capable of inhibiting in vitro hematopoiesis has been shown in aplastic anemia (AA), although whether such inhibition is mediated by a specific immune reaction involving an HLA allele remained unknown. We isolated a CD4+ Vbeta21+ T-cell clone that was most dominant among Vbeta21+ T cells in the bone marrow (BM) of an AA patient whose HLA-DRB1 alleles included 1501 and 0405. The T-cell clone named NT4.2 lysed an autologous Epstein-Barr virus-transformed lymphoblastoid cell line (LCL) and phytohemagglutinin-stimulated lymphocytes (PHA-blasts) as well as allogeneic LCLs sharing HLA-DRB1*0405. Cytotoxicity against LCL cells and PHA-blasts by NT4.2 was blocked by anti-HLA-DR monoclonal antibody (MoAb) or anti-CD3 MoAb. NT4.2 also lysed autologous BM mononuclear cells enriched with CD34+ cells that had been cultured for one week in the presence of colony-stimulating factors as well as allogeneic CD34+ cells of a normal individual carrying HLA-DRB1*0405, cultured in the same way. Moreover, NT4.2 strongly inhibited colony formation by hematopoietic progenitor cells derived from cultured CD34+ cells sharing HLA-DRB1*0405. These results indicate that the AA patient has T cells capable of killing hematopoietic cells in an HLA-DRB1*0405-restricted manner and that such cytotoxic T cells may contribute to the pathogenesis of AA.

  10. Opioid modulation of immunocompetence: Receptor characterization and second messenger involvement

    Energy Technology Data Exchange (ETDEWEB)

    Hemmick, L.M.

    1989-01-01

    The purpose of this thesis was to examine the effects of opioids on several indices of immunocompetence, determined the receptor specificity of these effects, and ascertain whether the actions of opioids on lymphocytes could be correlated with activation of second messenger systems. By measuring {sup 45}Ca{sup 2+} uptake into lymphocytes, it was demonstrated that {beta}-endorphin 1-31 ({beta}-END 1-31) enhanced rat thymocyte Ca{sup 2+} uptake in response to concanavalin A (Con A) but not phytohemagglutinin (PHA). Related opioid peptides and alkaloids were unable to mimic the effect, and naloxone did not block it, suggesting that {beta}-END 1-31 acted by binding to specific, non-opioid receptors on the thymocytes. Rat splenocyte Con A-stimulated Ca{sup 2+} uptake was not affected by {beta}-END 1-31. {beta}-END 1-31 did not affect basal Ca{sup 2+} uptake by either cell type. Using ({sup 3}H)thymidine uptake as an index of lymphocyte proliferation, {beta}-END 1-31 and several related opioid peptides reversed prostaglandin E{sub 1} (PGE{sub 1}) suppression of rat lymph node cell Con A- and PHA-stimulated proliferation. Naloxone did not block the reversal. {beta}-END 1-31 was unable to reverse forskolin and cholera toxin suppression of proliferation, indicating that the lowering of cyclic AMP levels was not the mechanism involved. Verapamil inhibition of proliferation was also not reversed by {beta}-END 1-31, suggesting that promotion of Ca{sup 2+} influx was not a major mechanism involved.

  11. Transplantation Outcomes for Severe Combined Immunodeficiency, 2000–2009

    Science.gov (United States)

    Pai, Sung-Yun; Logan, Brent R.; Griffith, Linda M.; Buckley, Rebecca H.; Parrott, Roberta E.; Dvorak, Christopher C.; Kapoor, Neena; Hanson, Imelda C.; Filipovich, Alexandra H.; Jyonouchi, Soma; Sullivan, Kathleen E.; Small, Trudy N.; Burroughs, Lauri; Skoda-Smith, Suzanne; Haight, Ann E.; Grizzle, Audrey; Pulsipher, Michael A.; Chan, Ka Wah; Fuleihan, Ramsay L.; Haddad, Elie; Loechelt, Brett; Aquino, Victor M.; Gillio, Alfred; Davis, Jeffrey; Knutsen, Alan; Smith, Angela R.; Moore, Theodore B.; Schroeder, Marlis L.; Goldman, Frederick D.; Connelly, James A.; Porteus, Matthew H.; Xiang, Qun; Shearer, William T.; Fleisher, Thomas A.; Kohn, Donald B.; Puck, Jennifer M.; Notarangelo, Luigi D.; Cowan, Morton J.; O’Reilly, Richard J.

    2014-01-01

    BACKGROUND The Primary Immune Deficiency Treatment Consortium was formed to analyze the results of hematopoietic-cell transplantation in children with severe combined immunodeficiency (SCID) and other primary immunodeficiencies. Factors associated with a good transplantation outcome need to be identified in order to design safer and more effective curative therapy, particularly for children with SCID diagnosed at birth. METHODS We collected data retrospectively from 240 infants with SCID who had received transplants at 25 centers during a 10-year period (2000 through 2009). RESULTS Survival at 5 years, freedom from immunoglobulin substitution, and CD3+ T-cell and IgA recovery were more likely among recipients of grafts from matched sibling donors than among recipients of grafts from alternative donors. However, the survival rate was high regardless of donor type among infants who received transplants at 3.5 months of age or younger (94%) and among older infants without prior infection (90%) or with infection that had resolved (82%). Among actively infected infants without a matched sibling donor, survival was best among recipients of haploidentical T-cell–depleted transplants in the absence of any pretransplantation conditioning. Among survivors, reduced-intensity or myeloablative pre-transplantation conditioning was associated with an increased likelihood of a CD3+ T-cell count of more than 1000 per cubic millimeter, freedom from immunoglobulin substitution, and IgA recovery but did not significantly affect CD4+ T-cell recovery or recovery of phytohemagglutinin-induced T-cell proliferation. The genetic subtype of SCID affected the quality of CD3+ T-cell recovery but not survival. CONCLUSIONS Transplants from donors other than matched siblings were associated with excellent survival among infants with SCID identified before the onset of infection. All available graft sources are expected to lead to excellent survival among asymptomatic infants. (Funded by the

  12. ANTI-INFLAMMATORY AND CYTOTOXICITY EFFECTS OF SALVADORA PERSICA (MESWAK EXTRACTS ON JURKAT T-CELLS

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    Farimah Sardari

    2015-04-01

    Full Text Available Salvadora persica (S. persica, Meswak, is an evergreen shrub to 6-7 m. It has many biological activities such as antipyretic, anti-inflammatory and antifungal activities. This study evaluated in vitro cytotoxic and anti-inflammatory effects of S. persica extracts on human oral Jurkat (T leukemia cells. Extracts from Meswak stick and leaves were tested in different concentrations for their cytotoxic and anti-inflammatory activities on human oral Jurkat T- cells. So treated cells viability with increasing concentrations of S. persica stick extract (0.008-0.2 μg/ml and leaves extract (0.016-0.5 μg/ml for 24, 48 or 72 hours was assessed by using the mitochondrial dependent reduction of yellow MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide to purple formazan. Also Enzyme-linked immunosorbent assay (ELISA was performed on supernatants from treated Jurkat T-cells with phytohemagglutinin (PHA and both extracts to quantify IL-6, IL-8 pro-inflammatory cytokines. Statistically significant differences were indicated by p <0.05. Incubation of Jurkat cells with sterile distilled water, negative control, didn't show any mortality through the incubation period. Against PHA, positive control, both stick and leaves extracts of S. persica like resulted in a dose-dependent decrease of IL-6 and IL-8 secretion (p <0.01. Although both extracts significantly inhibited survival of Jurkat cells (p < 0.01 in a dose- and time-dependent manner, stick extract exerted more cytotoxic effects on Jurkat cells than leaves extract of S. persica (p <0.03. In conclusion, although with increasing concentrations of both extracts anti-inflammatory properties were boosted, S. persica extracts had dose-dependent cytotoxic effects on human oral Jurkat T-cells.

  13. New immunological investigations on Helicobacter pylori-induced gastric ulcer in patients.

    Science.gov (United States)

    Rahimi, Hamid Reza; Rasouli, Manoochehr; Jamshidzadeh, Akram; Farshad, Shohreh; Firoozi, Mehdi Saberi; Taghavi, Ali Reza; Kiany, Simin

    2013-06-01

    Although Helicobacter pylori (Hp) plays an important role in the pathogenesis of chronic gastritis and gastric ulcer, little is known about the probable mechanisms of these types of gastrointestinal damage. To determine the precise mechanisms involved in ulcer formation, immune responses in patients with gastric ulcer (GUP) caused by Hp infection (Hp(+)) were compared with those of other gastritis patients (GP). The sensitivity and proliferation of peripheral blood mononuclear cells (PBMNCs) obtained from patients were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against exposure with complex Hp crude antigen (HPCA) and mitogen (phytohemagglutinin, PHA). Production of inflammatory cytokines, including interleukin (IL)-1β and IL-8, in serum and supernatants of PBMNCs were then measured by ELISA. It was found that, after stimulation with PHA, both IL-8 and IL-1β concentrations in sera and supernatants as well as proliferation and sensitivity were statistically greater in GUP Hp(+) than GP Hp(-) . Furthermore, HPCA inhibited the proliferation of PBMNCs dose-dependently; however, it stimulated IL-8 and IL-1β production in supernatants of mononuclear cells. Therefore, the up-regulated concentrations of IL-8 and IL-1β may have been caused by increase in the size of mononuclear cell subpopulations or in their cytokine secretory activity, indicating the greatest cell responsiveness in GUP Hp(+) patients. These results suggest that tissue damage and ulcers occur in patients who produce more IL-8 and IL-1β than patients who do not develop ulcers; the former consequently have more activated immune cells at the site of infection. Therefore, both host responses and Hp virulence factors may be involved in the development of gastric ulcers.

  14. Regulation of DNA synthesis and the cell cycle in human prostate cancer cells and lymphocytes by ovine uterine serpin

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    Hansen Peter J

    2008-01-01

    Full Text Available Abstract Background Uterine serpins are members of the serine proteinase inhibitor superfamily. Like some other serpins, these proteins do not appear to be functional proteinase inhibitors. The most studied member of the group, ovine uterine serpin (OvUS, inhibits proliferation of several cell types including activated lymphocytes, bovine preimplantation embryos, and cell lines for lymphoma, canine primary osteosarcoma and human prostate cancer (PC-3 cells. The goal for the present study was to evaluate the mechanism by which OvUS inhibits cell proliferation. In particular, it was tested whether inhibition of DNA synthesis in PC-3 cells involves cytotoxic actions of OvUS or the induction of apoptosis. The effect of OvUS in the production of the autocrine and angiogenic cytokine interleukin (IL-8 by PC-3 cells was also determined. Finally, it was tested whether OvUS blocks specific steps in the cell cycle using both PC-3 cells and lymphocytes. Results Recombinant OvUS blocked proliferation of PC-3 cells at concentrations as low as 8 μg/ml as determined by measurements of [3H]thymidine incorporation or ATP content per well. Treatment of PC-3 cells with OvUS did not cause cytotoxicity or apoptosis or alter interleukin-8 secretion into medium. Results from flow cytometry experiments showed that OvUS blocked the entry of PC-3 cells into S phase and the exit from G2/M phase. In addition, OvUS blocked entry of lymphocytes into S phase following activation of proliferation with phytohemagglutinin. Conclusion Results indicate that OvUS acts to block cell proliferation through disruption of the cell cycle dynamics rather than induction of cytotoxicity or apoptosis. The finding that OvUS can regulate cell proliferation makes this one of only a few serpins that function to inhibit cell growth.

  15. Effects of Space Missions on the Human Immune System: A Meta-Analysis

    Science.gov (United States)

    Greenleaf, J. E.; Barger, L. K.; Baldini, F.; Huff, D.

    1995-01-01

    Future spaceflight will require travelers to spend ever-increasing periods of time in microgravity. Optimal functioning of the immune system is of paramount importance for the health and performance of these travelers. A meta-analysis statistical procedure was used to analyze immune system data from crew members in United States and Soviet space missions from 8.5 to 140 days duration between 1968 and 1985. Ten immunological parameters (immunoglobulins A, G, M, D, white blood cell (WBC) count, number of lymphocytes, percent total lymphocytes, percent B lymphocytes, percent T lymphocytes, and lymphocyte reactivity to mitogen) were investigated using multifactorial, repeated measure analysis of variance. With the preflight level set at 100, WBC count increased to 154 +/- 14% (mean +/- SE; p less than or equal to 0.05) immediately after flight; there was a decrease in lymphocyte count (83 +/- 4%; p less than or equal to 0.05) and percent of total lymphocytes (69 +/- 1%; p less than or equal to 0.05) immediately after flight, with reduction in RNA synthesis to phytohemagglutinin (PHA) to 51 +/- 21% (p less than or equal to 0.05) and DNA synthesis to PHA to 61 +/- 8% (p less than or equal to 0.05) at the first postflight measurement. Thus, some cellular immunological functions are decreased significantly following spaceflight. More data are needed on astronauts' age, aerobic power output, and parameters of their exercise training program to determine if these immune system responses are due solely to microgravity exposure or perhaps to some other aspect of spaceflight.

  16. Immunoregulatory effects on T lymphocytes by human mesenchymal stromal cells isolated from bone marrow, amniotic fluid, and placenta.

    Science.gov (United States)

    Mareschi, Katia; Castiglia, Sara; Sanavio, Fiorella; Rustichelli, Deborah; Muraro, Michela; Defedele, Davide; Bergallo, Massimiliano; Fagioli, Franca

    2016-02-01

    Mesenchymal stromal cells (MSCs) are a promising tool in cell therapies because of their multipotent, bystander, and immunomodulatory properties. Although bone marrow represents the main source of MSCs, there remains a need to identify a stem cell source that is safe and easily accessible and yields large numbers of cells without provoking debates over ethics. In this study, MSCs isolated from amniotic fluid and placenta were compared with bone marrow MSCs. Their immunomodulatory properties were studied in total activated T cells (peripheral blood mononuclear cells) stimulated with phytohemagglutinin (PHA-PBMCs). In particular, an in vitro co-culture system was established to study: (i) the effect on T-lymphocyte proliferation; (ii) the presence of T regulatory lymphocytes (Treg); (iii) the immunophenotype of various T subsets (Th1 and Th2 naïve, memory, effector lymphocytes); (iv) cytokine release and master gene expression to verify Th1, Th2, and Th17 polarization; and (v) IDO production. Under all co-culture conditions with PHA-PBMCs and MSCs (independently of tissue origin), data revealed: (i) T proliferation inhibition; (ii) increase in naïve T and decrease in memory T cells; (iii) increase in T regulatory lymphocytes; (iv) strong Th2 polarization associated with increased interleukin-10 and interleukin-4 levels, Th1 inhibition (significant decreases in interleukin-2, tumor necrosis factor-α, interferon-γ, and interleukin-12) and Th17 induction (production of high concentrations of interleukins-6 and -17); (v) indoleamine-2,3-dioxygenase mRNA induction in MSCs co-cultured with PHA-PBMCs. AF-MSCs had a more potent immunomodulatory effect on T cells than BM-MSCs, only slightly higher than that of placenta MSCs. This study indicates that MSCs isolated from fetal tissues may be considered a good alternative to BM-MSCs for clinical applications.

  17. Use of retroviral-mediated gene transfer to deliver and test function of chimeric antigen receptors in human T-cells

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    Ana C. Parente-Pereira

    2014-07-01

    Full Text Available Chimeric antigen receptors (CARs are genetically delivered fusion molecules that elicit T-cell activation upon binding of a native cell surface molecule. These molecules can be used to generate a large number of memory and effector T-cells that are capable of recognizing and attacking tumor cells. Most commonly, stable CAR expression is achieved in T-cells using retroviral vectors. In the method described here, retroviral vectors are packaged in a two-step procedure. First, H29D human retroviral packaging cells (a derivative of 293 cells are transfected with the vector of interest, which is packaged transiently in vesicular stomatitis virus (VSV G pseudotyped particles. These particles are used to deliver the vector to PG13 cells, which achieve stable packaging of gibbon ape leukaemia virus (GALV-pseudotyped particles that are suitable for infection of human T-cells. The key advantage of the method reported here is that it robustly generates polyclonal PG13 cells that are 100% positive for the vector of interest. This means that efficient gene transfer may be repeatedly achieved without the need to clone individual PG13 cells for experimental pre-clinical testing. To achieve T-cell transduction, cells must first be activated using a non-specific mitogen. Phytohemagglutinin (PHA provides an economic and robust stimulus to achieve this. After 48-72 h, activated T-cells and virus-conditioned medium are mixed in RetroNectin-coated plasticware, which enhances transduction efficiency. Transduced cells are analyzed for gene transfer efficiency by flow cytometry 48 h following transduction and may then be tested in several assays to evaluate CAR function, including target-dependent cytotoxicity, cytokine production and proliferation.

  18. A new point mutation in the deoxyribonuclic acid-binding domain of the vitamine D receptor in a kindred with hereditary 1,25-dihydroxyvitamin d-resistant rickets

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    Yagi, Hideki; Miyake, Hiroshi; Nagashima, Kanji; Kuroume, Takayoshi (Gunma Univ. School of Medicine (Japan)); Ozone, K.; Pike, J.W. (Baylor College of Medicine, Houston, TX (United States))

    1993-02-01

    Hereditary 1,25-dihydroxyvitamin D [1,25-(OH)[sub 2]D]-resistant rickets (HVDRR) is a rare disorder characterized by rickets, alopecia, hypocalcemia, secondary hyperparathyroidism, and normal or elevated serum 1,25-dihydroxyvitamin D levels. The authors describe a patient with typical clinical characteristics of HVDRR, except that elevated levels of serum phosphorus were present coincident with increased levels of serum intact PTH. The patient was treated with high dose calcium infusion after an ineffective treatment with 1[alpha]-hydroxyvitamin D[sub 3]; serum calcium and phosphorus as well as intact PTH and alkaline phosphatase levels were normalized. Evaluation of phytohemagglutinin-activated lymphocytes derived from this patient revealed that 1,25-(OH)[sub 2]D[sub 3] was unable to inhibit thymidine incooperation, a result that contrast with the capacity of 1,25-(OH)[sub 2]D[sub 3] to inhibit uptake into normal activated lymphocytes. 1,25-(OH)[sub 2]D[sub 3] did not induce human osteocalcin promoter activity after transfection of this DNA linked to a reporter gene into patient cells. Cointroduction of a human vitamin D receptor (VDR) cDNA expression vector with the reporter plasmid, however, restored the hormone response. Evaluation of extracts from the patient cells for VDR DNA binding revealed a defect in DNA binding. Analysis of genomic DNA from the patient's cells by PCR confirmed the presence of a point mutation in exon 2 of the VDR. This exon directs synthesis of a portion of the DNA-binding domain of the receptor. We conclude that the genetic basis for 1,25-(OH)[sub 2]D[sub 3] resistance in this kindred with VDR-positive HVDRR is due to a single base mutation in the VDR that leads to production of a receptor unable to interact appropriately with DNA. 20 refs., 3 figs., 1 tab.

  19. α-Amylase inhibitor-1 gene from Phaseolus vulgaris expressed in Coffea arabica plants inhibits α-amylases from the coffee berry borer pest

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    Oliveira-Neto Osmundo B

    2010-06-01

    Full Text Available Abstract Background Coffee is an important crop and is crucial to the economy of many developing countries, generating around US$70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei, is responsible for worldwide annual losses of around US$500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an α-amylase inhibitor gene (α-AI1, which confers resistance against the coffee berry borer insect-pest, into C. arabica plants. Results We transformed C. arabica with the α-amylase inhibitor-1 gene (α-AI1 from the common bean, Phaseolus vulgaris, under control of the seed-specific phytohemagglutinin promoter (PHA-L. The presence of the α-AI1 gene in six regenerated transgenic T1 coffee plants was identified by PCR and Southern blotting. Immunoblotting and ELISA experiments using antibodies against α-AI1 inhibitor showed a maximum α-AI1 concentration of 0.29% in crude seed extracts. Inhibitory in vitro assays of the α-AI1 protein against H. hampei α-amylases in transgenic seed extracts showed up to 88% inhibition of enzyme activity. Conclusions This is the first report showing the production of transgenic coffee plants with the biotechnological potential to control the coffee berry borer, the most important insect-pest of crop coffee.

  20. Formocresol mutagenicity following primary tooth pulp therapy: an in vivo study.

    Science.gov (United States)

    Zarzar, P A; Rosenblatt, A; Takahashi, C S; Takeuchi, P L; Costa Júnior, L A

    2003-09-01

    To investigate whether formocresol, in Buckley's original formulation, is mutagenic in vivo to lymphocyte cultures obtained from the peripheral blood of children aged from 5 to 10 years old. These children were recruited from those attending the dental clinics of Recife City Council and the University of Pernambuco School of Dentistry, Brazil. The sample comprised 20 children who had primary teeth with cariously exposed vital pulps. Two venous blood samples were collected (6-8 ml) from each child, the first prior to vital pulpotomy (control group) and the second 24 h after pulpotomy (treated group). This research is a case-control study. The peripheral lymphocytes were grown in a complete culture medium consisting of 78% RPMI 1640 medium (a), supplemented with streptomycin (0.01 mg/ml), penicillin (0.005 ml(-1)), 20% fetal bovine serum (b) and 2% phytohemagglutinin (c). The lymphocytes were assessed for chromosomal aberrations via a previously published method which was modified. The cytogenetic analysis was performed in a blind test, where the slides were codified by an annotator and the scorers did not know which group they were analyzing. For each sample, this envolved the analysis of 200 metaphases. The level of significance adopted in the statistical test was 5.0% (pformocresol was mutagenic for one patient, raising doubt about the desirability of its use for pulpotomies in children. The results revealed that, from a statistical standpoint, formocresol is not mutagenic. However, further investigations are required, preferably with a larger sample, in patients needing more than one pulpotomy in order to observe whether an increase in the quantity of the drug would increase the quantity of chromosome aberrations and also to verify individual susceptibility to chromosome alterations with the use of formocresol.

  1. Transcriptomic Characterization of Innate and Acquired Immune Responses in Red-Legged Partridges (Alectoris rufa: A Resource for Immunoecology and Robustness Selection.

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    Natalia Sevane

    Full Text Available Present and future challenges for wild partridge populations include the resistance against possible disease transmission after restocking with captive-reared individuals, and the need to cope with the stress prompted by new dynamic and challenging scenarios. Selection of individuals with the best immune ability may be a good strategy to improve general immunity, and hence adaptation to stress. In this study, non-infectious challenges with phytohemagglutinin (PHA and sheep red blood cells allowed the classification of red-legged partridges (Alectoris rufa according to their overall immune responses (IR. Skin from the area of injection of PHA and spleen, both from animals showing extreme high and low IR, were selected to investigate the transcriptional profiles underlying the different ability to cope with pathogens and external aggressions. RNA-seq yielded 97 million raw reads from eight sequencing libraries and approximately 84% of the processed reads were mapped to the reference chicken genome. Differential expression analysis identified 1488 up- and 107 down-regulated loci in individuals with high IR versus low IR. Partridges displaying higher innate IR show an enhanced activation of host defence gene pathways complemented with a tightly controlled desensitization that facilitates the return to cellular homeostasis. These findings indicate that the immune system ability to respond to aggressions (either diseases or stress produced by environmental changes involves extensive transcriptional and post-transcriptional regulations, and expand our understanding on the molecular mechanisms of the avian immune system, opening the possibility of improving disease resistance or robustness using genome assisted selection (GAS approaches for increased IR in partridges by using genes such as AVN or BF2 as markers. This study provides the first transcriptome sequencing data of the Alectoris genus, a resource for molecular ecology that enables integration

  2. Impact of lithium alone and in combination with antidepressants on cytokine production in vitro.

    Science.gov (United States)

    Petersein, Charlotte; Sack, Ulrich; Mergl, Roland; Schönherr, Jeremias; Schmidt, Frank M; Lichtblau, Nicole; Kirkby, Kenneth C; Bauer, Katrin; Himmerich, Hubertus

    2015-01-01

    Lithium is an important psychopharmacological agent for the treatment of unipolar as well as bipolar affective disorders. Lithium has a number of side effects such as hypothyroidism and aggravation of psoriasis. On the other hand, lithium has pro-inflammatory effects, which appear beneficial in some disorders associated with immunological deficits, such as human immunodeficiency virus (HIV) infection and systemic lupus erythematosus (SLE). Therefore, immunological characteristics of lithium may be an important consideration in individualized therapeutic decisions. We measured the levels of the cytokines interleukin (IL)-1ß, IL-2, IL-4, IL-6, IL-22, IL-17 and tumour necrosis factor (TNF)-α in the stimulated blood of thirty healthy subjects supplemented with lithium alone, the antidepressants citalopram, escitalopram or mirtazapine alone, the combination of each antidepressant with lithium, and a no drug control. These drugs were tested under three blood stimulant conditions: murine anti-human CD3 monoclonal antibody OKT3 and the 5C3 monoclonal antibody (OKT3/5C3), phytohemagglutinin (PHA), and unstimulated blood. Lithium, alone and in combination with any of the tested antidepressants, led to a consistent increase of IL-1ß, IL-6 and TNF-α levels in the unstimulated as well as the stimulated blood. In the OKT3/5C3- and PHA-stimulated blood, IL-17 production was significantly enhanced by lithium. Lithium additionally increased IL-2 concentrations significantly in PHA-stimulated blood. The data support the view that lithium has pro-inflammatory properties. These immunological characteristics may contribute to side effects of lithium, but may also explain its beneficial effects in patients suffering from HIV infection or SLE.

  3. Effects of different levels of vitamin premix in finisher diets on performance, immuno-competence and meat lipid oxidation of chickens fed on corn-soybean meal.

    Science.gov (United States)

    Moravej, Hoseein; Alahyari-Shahrasb, Majid; Kiani, Ali; Bagherirad, Mona; Shivazad, Mahmood

    2013-01-01

    The present study was carried out to examine the effects of a vitamin premix (VP) reduction or withdrawal from finisher diet (29-43 days) on performance, immuno-competence, and characteristics of leg bones and meat lipid oxidation of chickens fed on corn-soybean meal based diet. A total of 900 male broiler chickens (Ross 308) were allocated to five treatment groups (0, 33%, 66%, 100% and 133% VP), with nine replicates per treatment group. At 29 and 36 days of ages, four birds from each replicate were injected with sheep red blood cells (SRBC). The cell-mediated immunity was determined via phytohemagglutinin (PHA) and 1-chloro 2-4-dinitrobenzen (DNCB) at 34 and 42 days of ages. At 33, 38 and 43 days of age, 42 days of ages, and two birds of each replicate were slaughtered and bone parameters measured. The oxidative stability was evaluated by thiobarbituric acid reactive substances (TBARS) on the thigh samples that were stored for 90 day at -80 ˚C. The results showed that reduction or withdrawal of VP from diets at different time points of the finisher period did not affect performance, immunocompetence and characteristics of leg bones. Results of TBARS showed that lipid peroxidation of the treatment without VP was significantly higher than of the other treatments when slaughtered at 43 days of age. Finally, the results of this study demonstrated that it is not possible to reduce the VP in finisher broilers' diets without negative effects on meat quality during the time of freezing.

  4. Chromosomal investigations in patients with mental retardation and/or congenital malformations

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    C.B. Santos

    2000-12-01

    Full Text Available We investigated the chromosomal constitution of patients with mental retardation and/or congenital malformations in order to determine genetic causes for such disturbances. The GTG and CBG banding patterns were studied using phytohemagglutinin M-stimulated lymphocytes cultured from peripheral blood. Among 98 individuals with mental retardation and/or congenital malformations who were analyzed there were 12 cases of Down's syndrome, two of Edward's syndrome, one of Patau's syndrome, five of Turner's syndrome, two of Klinefelter's syndrome, one of "cri-du-chat" syndrome, one case of a balanced translocation between chromosomes 13 and 14, one case of a derivative chromosome and one of a marker chromosome. We found abnormal chromosomes in 26% of the patients, 82% of which were numerical abnormalities, with the remaining 18% being structural variants. We conclude that patients with mental retardation and/or congenital malformations should be routinely karyotyped.Neste estudo investigamos a organização cromossômica de pacientes com retardo mental e/ou malformações congênitas, visando a avaliação de causas genéticas associadas a estes distúrbios. Os padrões de bandas GTG e CBG foram estudados a partir da cultura de linfócitos de sangue periférico, estimulados por fitohemaglutinina M. Dentre os 98 indivíduos portadores de retardo mental e/ou malformações congênitas analisados, diagnosticamos as seguintes síndromes: 12 casos de Down, dois de Edwards, um de Patau, cinco de Turner, dois de Klinefelter, um de "cri-du-chat", e um caso de translocação balanceada entre os cromossomos 13 e 14, um caso de cromossomo derivado e um outro de cromossomo marcador. Encontramos anomalias cromossômicas em 26% dos pacientes, das quais 82% eram alterações numéricas e o restante (18% representou rearranjos estruturais. Este percentual significativo enfatiza o uso da cariotipagem de rotina em pacientes com retardo mental e/ou malformações congênitas.

  5. Paroxetine and bupropion have no in vitro effects on lynphocyte proliferation and viability Paroxetina e bupropiona não apresentam efeito na viabilidade nem na proliferação de linfócitos in vitro

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    Ramiro Ronchetti

    2007-01-01

    Full Text Available OBJECTIVE: Initial studies with tricyclic antidepressants demonstrated that they jeopardize the immune system activity. Recent studies suggested that selective serotonin reuptake inhibitors would have stimulating immunological effects. Here, we explored the in vitro immunological effects of two antidepressants used in clinical practice, paroxetine (selective serotonin reuptake inhibitor and bupropion (norepinephrine and dopamine reuptake inhibitor. METHOD: Peripheral blood samples were obtained from 16 healthy volunteers and the peripheral blood mononuclear cells were isolated and cultured in vitro. We evaluated the effects of bupropion and paroxetine on cell viability as well as the ability to suppress phytohemagglutinin-induced lymphocyte proliferation. RESULTS: Both antidepressants produced neither significant effect on cell viability nor on T-cell proliferation. CONCLUSIONS: This could be of valuable information for the clinical practice when these drugs are administered. These results indicate a more favorable effect of such psychopharmacological drugs when compared to reported immunological effects associated with tryciclic antidepressants.OBJETIVO: Os estudos iniciais com antidepressivos tricíclicos demonstraram que estes prejudicam a atividade do sistema imune. Estudos mais recentes sugerem que os inibidores seletivos da recaptação de serotonina poderiam apresentar efeitos imunológicos estimulantes. No presente estudo, exploramos os efeitos imunológicos in vitro de dois antidepressivos usados na prática clínica, paroxetina (inibidor seletivo da recaptação de serotonina e bupropiona (inibidor da recaptação da noradrenalina e dopamina. MÉTODO: Obtiveram-se amostras de sangue periférico de 16 voluntários saudáveis e as células mononucleares do sangue periférico foram isoladas e cultivadas in vitro. Avaliamos os efeitos de bupropiona e da paroxetina em termos de viabilidade das células, como também a habilidade para

  6. Cell-type specific requirements for thiol/disulfide exchange during HIV-1 entry and infection.

    Science.gov (United States)

    Stantchev, Tzanko S; Paciga, Mark; Lankford, Carla R; Schwartzkopff, Franziska; Broder, Christopher C; Clouse, Kathleen A

    2012-12-03

    The role of disulfide bond remodeling in HIV-1 infection is well described, but the process still remains incompletely characterized. At present, the data have been predominantly obtained using established cell lines and/or CXCR4-tropic laboratory-adapted virus strains. There is also ambiguity about which disulfide isomerases/reductases play a major role in HIV-1 entry, as protein disulfide isomerase (PDI) and/or thioredoxin (Trx) have emerged as the two enzymes most often implicated in this process. We have extended our previous findings and those of others by focusing on CCR5-using HIV-1 strains and their natural targets--primary human macrophages and CD4+ T lymphocytes. We found that the nonspecific thiol/disulfide exchange inhibitor, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), significantly reduced HIV-1 entry and infection in cell lines, human monocyte-derived macrophages (MDM), and also phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC). Subsequent studies were performed using specific anti-PDI or Trx monoclonal antibodies (mAb) in HIV-1 envelope pseudotyped and wild type (wt) virus infection systems. Although human donor-to-donor variability was observed as expected, Trx appeared to play a greater role than PDI in HIV-1 infection of MDM. In contrast, PDI, but not Trx, was predominantly involved in HIV-1 entry and infection of the CD4+/CCR5+ T cell line, PM-1, and PHA-stimulated primary human T lymphocytes. Intriguingly, both PDI and Trx were present on the surface of MDM, PM-1 and PHA-stimulated CD4+ T cells. However, considerably lower levels of Trx were detected on freshly isolated CD4+ lymphocytes, compared to PHA-stimulated cells. Our findings clearly demonstrate the role of thiol/disulfide exchange in HIV-1 entry in primary T lymphocytes and MDM. They also establish a cell-type specificity regarding the involvement of particular disulfide isomerases/reductases in this process and may provide an explanation for differences

  7. Cell-type specific requirements for thiol/disulfide exchange during HIV-1 entry and infection

    Directory of Open Access Journals (Sweden)

    Stantchev Tzanko S

    2012-12-01

    Full Text Available Abstract Background The role of disulfide bond remodeling in HIV-1 infection is well described, but the process still remains incompletely characterized. At present, the data have been predominantly obtained using established cell lines and/or CXCR4-tropic laboratory-adapted virus strains. There is also ambiguity about which disulfide isomerases/ reductases play a major role in HIV-1 entry, as protein disulfide isomerase (PDI and/or thioredoxin (Trx have emerged as the two enzymes most often implicated in this process. Results We have extended our previous findings and those of others by focusing on CCR5-using HIV-1 strains and their natural targets - primary human macrophages and CD4+ T lymphocytes. We found that the nonspecific thiol/disulfide exchange inhibitor, 5,5'-dithiobis(2-nitrobenzoic acid (DTNB, significantly reduced HIV-1 entry and infection in cell lines, human monocyte-derived macrophages (MDM, and also phytohemagglutinin (PHA-stimulated peripheral blood mononuclear cells (PBMC. Subsequent studies were performed using specific anti-PDI or Trx monoclonal antibodies (mAb in HIV-1 envelope pseudotyped and wild type (wt virus infection systems. Although human donor-to-donor variability was observed as expected, Trx appeared to play a greater role than PDI in HIV-1 infection of MDM. In contrast, PDI, but not Trx, was predominantly involved in HIV-1 entry and infection of the CD4+/CCR5+ T cell line, PM-1, and PHA-stimulated primary human T lymphocytes. Intriguingly, both PDI and Trx were present on the surface of MDM, PM-1 and PHA-stimulated CD4+ T cells. However, considerably lower levels of Trx were detected on freshly isolated CD4+ lymphocytes, compared to PHA-stimulated cells. Conclusions Our findings clearly demonstrate the role of thiol/disulfide exchange in HIV-1 entry in primary T lymphocytes and MDM. They also establish a cell-type specificity regarding the involvement of particular disulfide isomerases/reductases in this

  8. Nanonized black soybean enhances immune response in senescence-accelerated mice

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    Yin-Ching Chan

    2009-02-01

    phytohemagglutinin-stimulated PBMC (P<0.05. However at higher concentrations (>50 μM, daidzein only reduced IL-10 and IFN-γ levels, whereas genistein reduced levels of the IL-2, IL-4, IL-10, IFN-γ mRNA and protein and these results suggest that the Nano-soy supplementation improved immune response in SAMP8 mice which may be attributable to higher daidzin content in the black soybean preparation.Keywords: nanonized black soybean, cytokines, senescence accelerated mice, splenocytes, peripheral blood mononuclear cells

  9. Improved performance and immunological responses as the result of dietary genistein supplementation of broiler chicks.

    Science.gov (United States)

    Rasouli, E; Jahanian, R

    2015-09-01

    The present study aimed to investigate the effect of supplemental genistein (an isoflavonoid) on performance, lymphoid organs' development, and cellular and humoral immune responses in broiler chicks. A total of 675-day-old male broiler chicks (Ross 308) were randomly assigned to the five replicate pens (15 chicks each) of nine experimental diets. Dietary treatments included a negative (not-supplemented) control diet, two positive control groups (virginiamycin or zinc-bacitracin, 20 mg/kg), and diets containing 10, 20, 40, 80, 160 and 320 mg/kg of genistein. The cutaneous basophil hypersensivity (CBH) test was measured at day 10 of age after toe web injection with phytohemagglutinin-P. In addition, sera samples were collected after different antigen inoculations to investigate antibody responses. At day 28 of age, three randomly selected birds from each pen were euthanized to evaluate the relative weights of lymphoid organs. Results showed that dietary supplementation of both antibiotics increased (P<0.01) feed intake during 1 to 42 days of age. Furthermore, daily weight gain was influenced (P<0.01) by dietary treatments throughout the trial, so that the birds fed on antibiotics and 20 to 80 mg/kg genistein diets revealed the greater weight gains compared with other experimental groups. The best (P<0.05) feed conversion ratio assigned to the birds fed on diets containing antibiotics and moderate levels (40 to 80 mg/kg) of genistein. Although the relative weights of thymus (P<0.05) and bursa of Fabricius (P<0.01) were greater in birds fed on genistein-supplemented diets compared with antibiotics-supplemented birds, the spleen weight was not affected by experimental diets. Similarly, CBH response and antibody titers against Newcastle and infectious bronchitis disease viruses were markedly (P<0.05) greater in chicks fed on diets supplemented with 20 to 80 mg/kg of genistein. Interestingly, the higher dosages of genistein suppressed CBH and antibody responses to the

  10. Genomic Analysis of Storage Protein Deficiency in Genetically Related Lines of Common Bean (Phaseolus vulgaris)

    Science.gov (United States)

    Pandurangan, Sudhakar; Diapari, Marwan; Yin, Fuqiang; Munholland, Seth; Perry, Gregory E.; Chapman, B. Patrick; Huang, Shangzhi; Sparvoli, Francesca; Bollini, Roberto; Crosby, William L.; Pauls, Karl P.; Marsolais, Frédéric

    2016-01-01

    A series of genetically related lines of common bean (Phaseolus vulgaris L.) integrate a progressive deficiency in major storage proteins, the 7S globulin phaseolin and lectins. SARC1 integrates a lectin-like protein, arcelin-1 from a wild common bean accession. SMARC1N-PN1 is deficient in major lectins, including erythroagglutinating phytohemagglutinin (PHA-E) but not α-amylase inhibitor, and incorporates also a deficiency in phaseolin. SMARC1-PN1 is intermediate and shares the phaseolin deficiency. Sanilac is the parental background. To understand the genomic basis for variations in protein profiles previously determined by proteomics, the genotypes were submitted to short-fragment genome sequencing using an Illumina HiSeq 2000/2500 platform. Reads were aligned to reference sequences and subjected to de novo assembly. The results of the analyses identified polymorphisms responsible for the lack of specific storage proteins, as well as those associated with large differences in storage protein expression. SMARC1N-PN1 lacks the lectin genes pha-E and lec4-B17, and has the pseudogene pdlec1 in place of the functional pha-L gene. While the α-phaseolin gene appears absent, an approximately 20-fold decrease in β-phaseolin accumulation is associated with a single nucleotide polymorphism converting a G-box to an ACGT motif in the proximal promoter. Among residual lectins compensating for storage protein deficiency, mannose lectin FRIL and α-amylase inhibitor 1 genes are uniquely present in SMARC1N-PN1. An approximately 50-fold increase in α-amylase inhibitor like protein accumulation is associated with multiple polymorphisms introducing up to eight potential positive cis-regulatory elements in the proximal promoter specific to SMARC1N-PN1. An approximately 7-fold increase in accumulation of 11S globulin legumin is not associated with variation in proximal promoter sequence, suggesting that the identity of individual proteins involved in proteome rebalancing might

  11. The method validation step of biological dosimetry accreditation process

    Energy Technology Data Exchange (ETDEWEB)

    Roy, L.; Voisin, P.A.; Guillou, A.C.; Busset, A.; Gregoire, E.; Buard, V.; Delbos, M.; Voisin, Ph. [Institut de Radioprotection et de Surete Nucleaire, LDB, 92 - Fontenay aux Roses (France)

    2006-07-01

    One of the missions of the Laboratory of Biological Dosimetry (L.D.B.) of the Institute for Radiation and Nuclear Safety (I.R.S.N.) is to assess the radiological dose after an accidental overexposure suspicion to ionising radiation, by using radio-induced changes of some biological parameters. The 'gold standard' is the yield of dicentrics observed in patients lymphocytes, and this yield is converted in dose using dose effect relationships. This method is complementary to clinical and physical dosimetry, for medical team in charge of the patients. To obtain a formal recognition of its operational activity, the laboratory decided three years ago, to require an accreditation, by following the recommendations of both 17025 General Requirements for the Competence of Testing and Calibration Laboratories and 19238 Performance criteria for service laboratories performing biological dosimetry by cyto-genetics. Diagnostics, risks analysis were realized to control the whole analysis process leading to documents writing. Purchases, personnel department, vocational training were also included in the quality system. Audits were very helpful to improve the quality system. One specificity of this technique is that it is not normalized therefore apart from quality management aspects, several technical points needed some validations. An inventory of potentially influent factors was carried out. To estimate their real effect on the yield of dicentrics, a Placket-Burman experimental design was conducted. The effect of seven parameters was tested: the BUdr (bromodeoxyuridine), PHA (phytohemagglutinin) and colcemid concentration, the culture duration, the incubator temperature, the blood volume and the medium volume. The chosen values were calculated according to the uncertainties on the way they were measured i.e. pipettes, thermometers, test tubes. None of the factors has a significant impact on the yield of dicentrics. Therefore the uncertainty linked to their use was

  12. Similar to Those Who Are Breastfed, Infants Fed a Formula Containing 2'-Fucosyllactose Have Lower Inflammatory Cytokines in a Randomized Controlled Trial.

    Science.gov (United States)

    Goehring, Karen C; Marriage, Barbara J; Oliver, Jeffery S; Wilder, Julie A; Barrett, Edward G; Buck, Rachael H

    2016-12-01

    Evidence suggests that human milk oligosaccharides (HMOs) provide multiple benefits to infants, including prebiotic effects, gut maturation, antimicrobial activities, and immune modulation. Clinical intervention studies with HMOs are required to confirm these benefits in infants. Our objective was to investigate the effects of feeding formulas supplemented with the HMO 2'-fucosyllactose (2'-FL) on biomarkers of immune function in healthy term infants. We performed a substudy nested within a randomized, double-blind, controlled growth and tolerance study in healthy singleton infants (birth weight ≥2490 g) who were enrolled by 5 d of life and exclusively formula-fed (n = 317) or breastfed (n = 107) from enrollment to 4 mo of age. Formula-fed infants were randomly assigned to receive 1 of 3 formulas, all containing 2.4 g total oligosaccharides/L [control: galacto-oligosaccharides (GOS) only; experimental formulas: GOS + 0.2 or 1.0 g 2'-FL/L], and compared with a breastfed reference group. For this substudy, blood samples were drawn from infants at 6 wk of age (n = 31-42/group). Peripheral blood mononuclear cells (PBMCs) were isolated for cellular phenotyping and stimulated ex vivo with phytohemagglutinin for proliferation and cell cycle progression or respiratory syncytial virus (RSV). Cytokine concentrations were measured in plasma and in ex vivo-stimulated culture supernatants. Breastfed infants and infants fed either of the experimental formulas with 2'-FL were not different but had 29-83% lower concentrations of plasma inflammatory cytokines than did infants fed the control formula [interleukin (IL) receptor antagonist (IL-1ra), IL-1α, IL-1β, IL-6, and tumor necrosis factor α (TNF-α)] (P ≤ 0.05). In ex vivo RSV-stimulated PBMC cultures, breastfed infants were not different than either of the groups fed formula with 2'-FL, but they had lower concentrations of TNF-α (31%) and interferon γ (IFN-γ 54%) (P ≤ 0.05) and tended to have lower IL-1ra (25%) and

  13. Plasmid DNA as a special stimular to stimulate lymphocyte proliferation%DNA作为一种特异性刺激剂刺激淋巴细胞增殖的研究

    Institute of Scientific and Technical Information of China (English)

    吴琼; 孙英军; 张艳; 郑海学

    2011-01-01

    目的 为了探讨质粒DNA体外刺激淋巴细胞的增殖状况,建立了一种方便可靠的评价豚鼠细胞免疫水平的试验方法.方法 用羧基荧光素乙酰乙酸琥珀酰亚胺酯(cFsEl染色豚鼠全血,经植物血凝素(PHA)和质粒DNA刺激培养,利用流式细胞术分析细胞的增殖状况.结果 豚鼠全血经PHA和DNA刺激,淋巴细胞增殖能力不同;未免疫组经DNA和PHA刺激后增殖的差异显著,免疫组差异不显著.DNA质粒在体内外均可作为刺激源刺激淋巴细胞增殖.结论 建立了一种基于活细胞染料CFSE染色的豚鼠全血淋巴细胞增殖试验方法,可方便、快速、有效地评价细胞免疫水平.%To investigate the level of lymphocyte proliferation stimulated by plasmid DNA in vitro, and to establish a convenient and reliable method to assess the level of cellular immunity in guinea pig, the whole blood of guinea pig was stained by Carboxyfluorescein diacetate succinimidyl ester (CFSE), then stimulated by phytohemagglutinin(PHA) and plasmid DNA, and cultivated for 3 days.The cell proliferation was detected by flow cytometry.We observed that PHA and DNA stimulation could promote lymphocyte proliferation: the proliferation in non-immune group was significantly enhanced, while the inactivated vaccine immune group did not significantly changed, which indicate that plasmid DNA can be used as animmunogen to stimulate lymphocyte proliferation.Through this study, a live cell-based dye CFSE staining of guinea pig whole blood lymphocyte proliferation test method was established to evaluate the cellular immunity conveniently and effectively.

  14. Atividades biológicas das lectinas PHA, WGA, jacalina e artocarpina Biological activities of PHA, WGA, jacalin and artocarpin lectins

    Directory of Open Access Journals (Sweden)

    Ana Maria Sell

    2000-05-01

    Full Text Available As lectinas são (glicoproteínas que se ligam a açúcares. O interesse destas moléculas na investigação científica se deve às diversas atividades biológicas a elas atribuídas. Além da identificação de grupos sangüíneos, da caracterização de microorganismos e da estimulação mitogênica de células imunes, as lectinas estão envolvidas nos processos de reconhecimento e das interações celulares. No presente trabalho, revisamos algumas atividades biológicas das lectinas PHA (fitohemaglutinina de Phaseolus vulgaris, WGA (aglutinina de germe de trigo, jacalina e artocarpina (lectinas de Artocarpus integrifolia. PHA, jacalina e artocarpina são mitogênicas para os linfócitos, estimulam a produção de citocinas endógenas, a atração e a ativação de leucócitos. WGA é uma lectina não-mitogênica, no entanto, modula a resposta de defesa estimulando a liberação de superóxido pelos neutrófilos. O entendimento das atividades biológicas permitirá o uso destas moléculas como uma ferramenta útil no diagnóstico e tratamento de muitas doenças.Lectins are ubiquitous (glycoproteins, which exhibit a specific and reversibly carbohydrate binding activity. Because of their structural complexity and variability, they can participate in many physiological functions. Besides the blood groups identification, microorganism characterization and mitogenic immune cell proliferation, they are involved in the cell recognition and cell-cell interaction. In the present work we revised some biological activities of the PHA (Phaseolus vulgaris phytohemagglutinin, WGA (wheat germ agglutinin, jacalin and artocarpin (Artocarpus integrifolia lectins. PHA, jacalin and artocarpin may stimulate lymphocyte proliferation, attract and activate mononuclear cells and stimulate endogenous citokines production. WGA is a nonmitogenic lectin, however it may stimulate neutrophil superoxide production. The understanding of the lectin biological activities may

  15. Kidney bean: a major sensitizer among legumes in asthma and rhinitis patients from India.

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    Ramkrashan Kasera

    Full Text Available BACKGROUND: The prevalence of IgE mediated food allergies has increased over the last two decades. Food allergy has been reported to be fatal in highly sensitive individuals. Legumes are important food allergens but their prevalence may vary among different populations. The present study identifies sensitization to common legumes among Indian population, characterizes allergens of kidney bean and establishes its cross reactivity with other legumes. METHODOLOGY: Patients (n = 355 with history of legume allergy were skin prick tested (SPT with 10 legumes. Specific IgE (sIgE and total IgE were estimated in sera by enzyme-linked immunosorbent assay. Characterization of kidney bean allergens and their cross reactivity was investigated by immunobiochemical methods. Identification of major allergens of kidney bean was carried out by mass spectrometry. PRINCIPAL FINDINGS: Kidney bean exhibited sensitization in 78 (22.0% patients followed by chickpea 65 (18.0% and peanut 53 (15%. SPT positive patients depicted significantly elevated sIgE levels against different legumes (r = 0.85, p<0.0001. Sera from 30 kidney bean sensitive individuals exhibited basophil histamine release (16-54% which significantly correlated with their SPT (r = 0.83, p<0.0001 and sIgE (r = 0.99, p<0.0001. Kidney bean showed eight major allergens of 58, 50, 45, 42, 40, 37, 34 and 18 kDa on immunoblot and required 67.3±2.51 ng of homologous protein for 50% IgE inhibition. Inhibition assays revealed extensive cross reactivity among kidney bean, peanut, black gram and pigeon pea. nLC-MS/MS analysis identified four allergens of kidney bean showing significant matches with known proteins namely lectin (phytohemagglutinin, phaseolin, alpha-amylase inhibitor precursor and group 3 late embryogenesis abundant protein. CONCLUSION/SIGNIFICANCE: Among legumes, kidney bean followed by chick pea and peanut are the major allergic triggers in asthma and rhinitis patients in India

  16. A combined stress hormone infusion decreases in vivo protein synthesis in human T lymphocytes in healthy volunteers.

    Science.gov (United States)

    Januszkiewicz, A; Essén, P; McNurlan, M A; Ringdén, O; Garlick, P J; Wernerman, J

    2001-11-01

    In vivo protein synthesis decreases in mononuclear cells following a combined stress hormone infusion given to healthy volunteers as a human trauma model. Here, the purpose was to further investigate this finding and to measure in vivo protein synthesis in isolated T lymphocytes. Furthermore, the effects of stress hormones on the lymphocyte subpopulations and mononuclear cells, characterized by flow cytometry and phytohemagglutinin (PHA)-induced and unstimulated proliferative responses in vitro, were elucidated. Healthy volunteers (n = 16) were randomized into 2 groups to receive either a stress hormone or a saline infusion for 6 hours. In vivo protein synthesis was studied before and after the treatment by measuring the incorporation of stable isotopically-labeled phenylalanine into lymphocyte and mononuclear cell proteins. Protein synthesis decreased after stress hormone infusion in both cell populations: in T lymphocytes from 13.0% +/- 0.7%/d (mean +/- SD) to 8.6% +/- 2.1%/d (P <.01) and in mononuclear cells from 13.3% +/- 1.2%/d to 6.3 +/- 2.0%/d (P <.001). No change in proliferative responsiveness in vitro was observed. The stress hormone infusion produced a decrease in the percentage of T helper CD3/CD4 from 41% to 18% (P <.001), T cytotoxic CD3/CD8 from 27% to 15% (P <.001), as well as total T CD3 cells from 69% to 35% (P <.001). There was an increase in the percentage of natural killer (NK) cells CD16/CD56 from 17% to 55% (P <.001). Determination of phenotypes expressed on activated T lymphocytes showed that CD3/HLA-DR was unchanged and CD3/CD25 decreased from 14% to 7% (P <.01) in the stress hormone group. The study showed that the decrease of in vivo protein synthesis was 34% in T lymphocytes as compared with 53% in mononuclear cells, when determined immediately after a 6-hour stress hormone infusion. This change was associated with a pronounced decrease in all lymphocyte subpopulations, except for the NK cells, which increased substantially.

  17. Evaluation of Lymphocyte Transformation Test Results in Patients with Delayed Hypersensitivity Reactions following the Use of Anticonvulsant Drugs.

    Science.gov (United States)

    Karami, Zahra; Mesdaghi, Mehrnaz; Karimzadeh, Parvaneh; Mansouri, Mahboubeh; Taghdiri, Mohammad Mehdi; Kayhanidoost, Zarrintaj; Jebelli, Bita; Shekarriz Foumani, Reza; Babaie, Delara; Chavoshzadeh, Zahra

    2016-01-01

    Administration of the anticonvulsant drugs phenobarbital, phenytoin, carbamazepine and lamotrigine can be associated with severe hypersensitivity reactions. The lymphocyte transformation test (LTT) is a method to determine which drug has caused the hypersensitivity reaction. This study was done to evaluate the results of LTT in patients with delayed hypersensitivity reactions following the administration of anticonvulsants. Twenty-four patients with hypersensitivity reactions, e.g. drug-induced hypersensitivity syndrome/drug rash and eosinophilia with systemic symptoms (DIHS/DRESS), Stevens-Johnson syndrome (SJS) and toxic epidermal necrosis (TEN), following the administration of anticonvulsant drugs, and 24 patients who had used anticonvulsant drugs but did not have hypersensitivity reactions (the control group) were included in this study. Peripheral blood mononuclear cells were isolated. The cells were stimulated with the drugs, phytohemagglutinin as a mitogen and Candida as an antigen (positive controls). Lymphocyte proliferation was measured using the BrdU proliferation assay kit (Roche, Germany). The stimulation index was calculated as the mean ratio of the OD of stimulated cells divided by the OD of unstimulated cells. The results in the case and control groups were compared. Of 24 patients in the test group, 14 (58.3%) had positive LTT results and 10 (41.7%) had negative results. Among patients in the control group, 1 (4.2%) had a positive LTT result and 23 (95.8%) had negative results. Among the patients who had received carbamazepine and phenytoin, there was a significant difference between the results of LTT in the case and control groups (p = 0.002 and p = 0.028, respectively). Although patients receiving lamotrigine and phenobarbital had more positive LTT results in the case group than in the control group, these differences were not statistically significant. The sensitivity, specificity, positive predictive value and negative predictive value of LTT

  18. EVALUATION OF THE ANTITUMOR ACTIVITY OF HUMAN IL-4 BY IN VITRO AND IN V1VO ASSAYS

    Institute of Scientific and Technical Information of China (English)

    王彤钢; 陈慰峰

    1994-01-01

    The characteristics of rhuIL-4 induced cytotoxicity was detected in vitro by using 51 Cr release assay and the anti-tumor activity of rhuIL-4 induced killer cell was evaluated in vivo by using a human tumor model in nudemice.huIL-4 can induce LAK activity from peripheral blood lymphocytes(PBMC) stimulated with phytohemagglutinin(PHA).Compared with the LAK activity induced by rhuIL-2,the cytotoxicity of the killer cells induced by rhuIL-4 to K562 and Raji cells was lower ,but that to TBL-E,a human lymphoid leukemia cell line established in our laboratory,and PHA-activated blast cells(PHA-blasts) was of similar magnitude.In the cytotoxicity assay using PHA-blasts,the addition of PHA increased the IL-4 induced killer cell cytotoxicity by 131%,but had no effect on IL-2-induced ki8ller cell cytotoxicity.This implies that IL-4 mainly induces CTL-like activity,while IL-2 mainly induces NK-like activity,An experimental human tumor model in nude mice was established by injection of TBL-E human leukemia cells.The anti-tumor activity of rhuIL-4 was evaluated by injection of haman LAK cells induced from PHA-blasts by rhuIL-2+rhuIL-4 and human cytokines into tumor-bearing nude mice.The results showed that human LAK cells effectively inhibit the tumorigenicity of TBL-E cells in nude mice with an inhibition rate of 61%.The antitumor effect of rhuIL-2 was better than that of rIL-4 ,and the antitumor effect of rhuIL-2+rhuIL-4 was similar to that of rhuIL-2 ,though the former delayed the occurence of tumors.Our data imply the potential application of human IL-4 in clinic,and provide an animal model to evaluate the anti-tumor activity of human cytokine(s) with species specificity.

  19. Synthesis, biological evaluation, and molecular docking studies of benzyl, alkyl and glycosyl [2-(arylamino)-4,4-dimethyl-6-oxo-cyclohex-1-ene]carbodithioates, as potential immunomodulatory and immunosuppressive agents.

    Science.gov (United States)

    El Ashry, El Sayed H; Amer, Mohammad R; Abdalla, Omer M; Aly, Aly A; Soomro, Samreen; Jabeen, Almas; Halim, Sobia Ahsan; Ahmed Mesaik, M; Ul-Haq, Zaheer

    2012-05-01

    The immunomodulating properties of functionalized [2-(arylamino)-4,4-dimethyl-6-oxo-cyclohex-1-ene] carbodithioates and 6,6-dimethyl-4-(2-(propan-2-ylidene)hydrazinyl)-6,7-dihydro-2H-indazole-3(5H)-thione compounds have been investigated. Four of them, 13, 18, 19 and 20 inhibited PBMC proliferation induced by phytohemagglutinin (PHA) in a dose dependent manner with an IC(50) of ≤ 20 μM. The Th-1 cytokine, interleukin-2 (IL-2) in PHA/PMA-stimulated peripheral blood mononuclear cells (PBMCs) is significantly inhibited by 13, 19 and 20 with an IC(50) of 8.4 ± 0.4, 5.34 ± 0.15 and 4.9 ± 0.7 μM, respectively. They also inhibited the PMA/lipopolysaccharide-induced proinflammatory cytokines, IL-1β and TNF-α production in human monocytic leukemia cells (THP-1), by 86%, 46% and 59.2% for IL-1β and by 83.8%, 48.2% and 58.7% for TNF-α, respectively. Only 20 showed significant suppressive activity against the phagocyte oxidative burst in a dose dependent manner, with an IC(50) of 23.8 μM. LPS-induced nitrites in mouse macrophages were found to be inhibited by compounds 6, 8, 13-15 and 19 with an IC(50), which range between 7.7 and 63 μM. The cytotoxicity for the active compounds was also studied on Rat Wistar Hepatocyte cell line, CC1 and the Mouse Fibroblast cell line 3T3 NIH in the presence of compounds using a standard MTT assay. Furthermore, structural-activity relationship using automated docking software revealed that active compounds 7, 13 and 19, adapted the same binding mode, however the most active compound 20 is found deeply inserted within the ligand binding site of IL-2, as multiple hydrophobic and hydrophilic key interactions stabilize the compound inside the binding site, thus contributing higher activity.

  20. Evaluation of Th1 and Th17 cells cytokines in cell culture stimulated in women with recurrent spontaneous abortion

    Directory of Open Access Journals (Sweden)

    Y Varghaiyan

    2013-10-01

    Full Text Available Introduction: Various immunological abnormalities have been reported in women with RSA of unknown aetiologies including autoimmune abnormalities and increased cellular immunity such as elevated natural killer (NK , Th1 and Th17 cell levels. Th17 and Th1 cells play a central role during inflammation. Th1 cells product cytokines IFN-γ, IL-2 and Th17 cells mainly cytokines IL-17A, F, IL-22. The aim of this study is evaluation of Th1 and Th17 activity in women with recurrent spontaneous abortion. Methods: In this case-control study, 30 women with history of two or more abortion who at least 3 months past after last abortion considered as case group and 30 normal fertile healthy women with at least one delivery as control group. We determined the levels of IL-17A, F and IFN-γ in cell culture supernatant of peripheral blood mononuclear cells (PBMC stimulated with the mitogen phytohemagglutinin (PHA by enzyme-linked immunosorbent assay (ELISA method and compared in the two groups. The results obtained using the one-sample kolmogorov-smirnov Test, Kruskal-wallis Test and Spearman were analyzed using SPSS 16 software. Results: The level of IFN- γ in case group was significantly higher than control group (186/53±30/41 versus 88/06±21/44 pg/ml, P < 0.005. Also the level of IL-17 A, F in case group was significantly higher than control group (84/74±21/26 versus 28/41±8 pg/ml, P < 0.01. IFN-γ concentration showed positive correlation with IL-17 A, F in case group (P=0.015, r= 0.455. Conclusion: In this study the increased levels of cytokines IFN- γ and IL-17 A, F in women with recurrent spontaneous abortion shows a propensity of pro inflammation via Th17 and Th1 immunity and may be these cells play a pivotal role in rejecting fetus antigens.

  1. In vitro characterization of the immunotoxic potential of several perfluorinated compounds (PFCs)

    Energy Technology Data Exchange (ETDEWEB)

    Corsini, Emanuela, E-mail: emanuela.corsini@unimi.it [Laboratory of Toxicology, Department of Pharmacological Sciences, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano (Italy); Sangiovanni, Enrico [Laboratory of Pharmacognosy, Department of Pharmacological Sciences, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano (Italy); Avogadro, Anna; Galbiati, Valentina; Viviani, Barbara; Marinovich, Marina; Galli, Corrado L. [Laboratory of Toxicology, Department of Pharmacological Sciences, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano (Italy); Dell' Agli, Mario [Laboratory of Pharmacognosy, Department of Pharmacological Sciences, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano (Italy); Germolec, Dori R. [National Toxicology Program, National Institute of Environmental Health Sciences, NIH, RTP, NC (United States)

    2012-01-15

    We have previously shown that PFOA and PFOS directly suppress cytokine secretion in immune cells, with different mechanisms of action. In particular, we have demonstrated a role for PPAR-α in PFOA-induced immunotoxicity, and that PFOS has an inhibitory effect on LPS-induced I-κB degradation. These studies investigate the immunomodulatory effects of four other PFCs, namely PFBS, PFOSA, PFDA, and fluorotelomer using in vitro assays. The release of the pro-inflammatory cytokines IL-6 and TNF-α was evaluated in lipolysaccharide (LPS)-stimulated human peripheral blood leukocytes (hPBL) and in the human promyelocytic cell line THP-1, while the release of IL-10 and IFN-γ was evaluated in phytohemagglutinin (PHA)-stimulated hPBL. All PFCs suppressed LPS-induced TNF-α production in hPBL and THP-1 cells, while IL-6 production was suppressed by PFOSA, PFOS, PFDA and fluorotelomer. PFBS, PFOSA, PFOS, PFDA and fluorotelomer inhibited PHA-induced IL-10 release, while IFN-γ secretion was affected by PFOSA, PFOS, PFDA and fluorotelomer. Leukocytes obtained from female donors appear to be more sensitive to the in vitro immunotoxic effects of PFCs when their responses are compared to the results obtained using leukocytes from male donors. Mechanistic investigations demonstrated that inhibition of TNF-α release in THP-1 cells occurred at the transcriptional level. All PFCs, including PFOA and PFOS, decreased LPS-induced NF-κB activation. With the exception of PFOA, none of the PFCs tested was able to activate PPARα driven transcription in transiently transfected THP-1 cells, excluding a role for PPARα in the immunomodulation observed. PFBS and PFDA prevented LPS-induced I-κB degradation. Overall, these studies suggest that PFCs affect NF-κB activation, which directly suppresses cytokine secretion by immune cells. Our results indicate that PFOA is the least active of the PFCs examined followed by PFBS, PFDA, PFOS, PFOSA and fluorotelomer. -- Research Highlights: ► PFCs

  2. Sequencing and expression analysis of CD3γ/δ and CD3εchains in mandarin fish, Siniperca chuatsi

    Institute of Scientific and Technical Information of China (English)

    GUO Zheng; NIE Pin

    2013-01-01

    The genomic and cDNA sequences of the CD3γ/δ and CD3ε homologues in the mandarin fish,Siniperca chuatsi,were determined.As in other vertebrate CD3 molecules,the deduced amino acid sequences of mandarin fish CD3γ/δ and CD3ε contained conserved residues and motifs,such as cysteine residues and CXXC and immunoreceptor tyrosine-based activation motifs.However,mandarin fish CD3γ/δ and CD3ε showed some differences to their mammalian counterparts,specifically the absence of a negatively charged residue in the transmembrane region of CD3γ/δ.Additionally,while an N-glycosylation site was present in CD3ε,the site was not observed in CD3γ/δ.The CD3γ/δ and CD3ε subunit sequences contain six and five exons,respectively,consistent with homologues from Atlantic salmon,Salmo salar.Phylogenetic analysis also revealed that CD3γ/δ and CD3ε in mandarin fish are closely related to their counterparts in Acanthopterygian fish.Real-time PCR showed CD3γ/δ and CD3ε were expressed mainly in the thymus and spleen in normal healthy fish and,to a lesser extent,in mucosal-associated lymphoid tissues,such as the intestine and gills.When lymphocytes isolated from head kidney were treated with the mitogens phytohemagglutinin,concanavalin,and polyriboinosinic polyribocytidylic acid,mRNA expression levels of CD3γ/δ and CD3ε were significantly elevated within 12 h of treatment.This indicated the presence of T lymphocytes in the head kidney of teleost fish,and also the recognition of mitogens by the lymphocytes.Mandarin fish infected with the bacterial pathogen Flavobacterium columnare also showed an increase in the expression of CD3γ/δ and CD3ε mRNA,indicating that CD3γ/δ and CD3ε lymphocytes are involved in the immune response of this species.

  3. Sequencing and expression analysis of CD3γ/δ and CD3ɛ chains in mandarin fish, Siniperca chuatsi

    Science.gov (United States)

    Guo, Zheng; Nie, Pin

    2013-01-01

    The genomic and cDNA sequences of the CD3γ/δ and CD3ɛ homologues in the mandarin fish, Siniperca chuats i, were determined. As in other vertebrate CD3 molecules, the deduced amino acid sequences of mandarin fish CD3γ/δ and CD3ɛ contained conserved residues and motifs, such as cysteine residues and CXXC and immunoreceptor tyrosine-based activation motifs. However, mandarin fish CD3γ/δ and CD3ɛ showed some differences to their mammalian counterparts, specifically the absence of a negatively charged residue in the transmembrane region of CD3γ/δ. Additionally, while an N -glycosylation site was present in CD3ɛ, the site was not observed in CD3γ/δ. The CD3γ/δ and CD3ɛ subunit sequences contain six and five exons, respectively, consistent with homologues from Atlantic salmon, Salmo salar. Phylogenetic analysis also revealed that CD3γ/δ and CD3ɛ in mandarin fish are closely related to their counterparts in Acanthopterygian fish. Real-time PCR showed CD3γ/δ and CD3ɛ were expressed mainly in the thymus and spleen in normal healthy fish and, to a lesser extent, in mucosal-associated lymphoid tissues, such as the intestine and gills. When lymphocytes isolated from head kidney were treated with the mitogens phytohemagglutinin, concanavalin, and polyriboinosinic polyribocytidylic acid, mRNA expression levels of CD3γ/δ and CD3ɛ were significantly elevated within 12 h of treatment. This indicated the presence of T lymphocytes in the head kidney of teleost fish, and also the recognition of mitogens by the lymphocytes. Mandarin fish infected with the bacterial pathogen Flavobacterium columnare also showed an increase in the expression of CD3γ/δ and CD3ɛ mRNA, indicating that CD3γ/δ and CD3ɛ lymphocytes are involved in the immune response of this species.

  4. Reduced rate of DNA replication fork movement in megaloblastic anemia.

    Science.gov (United States)

    Wickremasinghe, R G; Hoffbrand, A V

    1980-01-01

    Chromatography on benzoylated naphthoylated DEAE-cellulose has been used to fractionate fully double-stranded from partially single-stranded DNA molecules. DNA was extracted from phytohemagglutinin-stimulated lymphocytes from patients with megaloblastic anemia resulting from vitamin B12 or folate deficiency after pulse-labeling the cells with [3H]thymidine for 5 min and chasing in unlabeled medium for 24 h. No gross accumulation of partially single-stranded material was observed in the DNA of these cells when compared with DNA from similarly labeled control cells obtained by the addition of 5-formyl tetrahydrofolic acid to the culture medium. When DNA from lymphocytes labeled with a 5-min pulse of [3H]thymidine and sheared to fragments of an average length of 18 micrometer was chromatographed on benzoylated naphthoylated DEAE-cellulose, approximately 80% of the label was recovered in the partially single-stranded fraction. After chasing in unlabeled medium the label was progressively transferred to the double-stranded fraction over a period of 2--3 h. The rate of transfer was slower in megaloblastic lymphocytes than in controls. The difference in rate suggested a slower rate of replication fork movement in megaloblastic lymphocytes and so the density shift technique of Painter and schaeffer (J. Mol. Biol. 45: 467--479, 1969) was used to measure the fork rate directly. [3H]Deoxycytidine was used as the labeled nucleoside to avoid possible complications arising from [3H]thymidine labeling of megaloblastic cells. Investigations on the lymphocytes from four patients showed that the replication fork rate in vitamin-treated control lyphocytes was about 1 micrometer/min. The fork rates in the corresponding untreated cells were invariably lower and rates ranging from 40 to 92% of those of controls were observed. Normal lymphocytes treated with the deoxynucleotide pool-depleting drugs methotrexate or hydroxyurea displayed defects in DNA synthesis similar to those of

  5. Alterations of cell-mediated immune response in children with febrile seizures Alterações da resposta imune celular em crianças portadoras de convulsão febril

    Directory of Open Access Journals (Sweden)

    Terezinha C.B. Montelli

    1997-06-01

    Full Text Available The aim of the present investigation was to study the distribution of T-cell subsets in peripheral blood defined by monoclonal antibodies and by the lymphocyte proliferative response to phytohemagglutinin (PH A in 30 children with febrile seizures and in 14 age-matched control subjects. Frequent respiratory, urinary and dermatologic infections were observed in 22 patients. The immunologic parameters showed that 64% of the patients presented an increased number of CD8+ cells and a low helper/suppressor ratio was observed in 60% of the patients. In addition, the proliferative response of lymphocytes to PHA was impaired in the patients. It was observed the presence of inhibitory activity on lymphocyte function in the plasma of 33% of children with febrile seizures. These results suggest that patients with febrile seizures have an impairment of cellular immunity that may be connected with this epileptic syndrome and explain the infections observed.O objetivo da presente investigação foi estudar a distribuição das subpopulações de células T por meio de anticorpos monoclonais e a resposta proliferativa de linfócitos em resposta a fito-hemaglutinina (PHA em 30 crianças portadoras de convulsão febril e em 14 crianças saudáveis de mesma faixa etária dos pacientes. Infecções respiratórias, urinárias e dermatológicas frequentes foram observadas em 22 pacientes. Os parâmetros imunológicos demonstraram que 64% dos pacientes apresentaram valores elevados de células CD8+. Diminuição da relação de células CD4/CD8 foi observada em 60% dos pacientes. Além disso, a resposta proliferativa de linfócitos frente a PHA apresentou-se deprimida nos pacientes. Foi observada a presença de atividade inibidora da função de linfócitos no plasma de 33% das crianças com convulsão febril. Esses resultados sugerem que pacientes com convulsão febril apresentam depressão da resposta imune celular que poderia implicar em associação patog

  6. Associations between immune function and air pollution among postmenopausal women living in the Puget Sound airshed

    Science.gov (United States)

    Williams, Lori A.

    Air pollution is associated with adverse health outcomes, and changes in the immune system may be intermediate steps between exposure and a clinically relevant adverse health outcome. We analyzed the associations between three different types of measures of air pollution exposure and five biomarkers of immune function among 115 overweight and obese postmenopausal women whose immunity was assessed as part of a year-long moderate exercise intervention trial. For air pollution metrics, we assessed: (1) residential proximity to major roads (freeways, major arterials and truck routes), (2) fine particulate matter(PM2.5) at the nearest monitor to the residence averaged over three time windows (3-days, 30-days and 60-days), and (3) nitrogen dioxide (NO2) modeled based on land use characteristics. Our immune biomarkers included three measures of inflammation---C-reactive protein, serum amyloid A and interleukin-6---and two measures of cellular immunity---natural killer cell cytotoxicity and T lymphocyte proliferation. We hypothesized that living near a major road, increased exposure to PM2.5 and increased exposure to NO2 would each be independently associated with increased inflammation and decreased immune function. We observed a 21% lower average natural killer cell cytotoxicity among women living within 150 meters of a major arterial road compared to other women. For PM2.5 , we observed changes in 3 of 4 indicators of lymphocyte proliferation stimulated by anti-CD3---an antibody to the T cell receptor associated with increases in 3-day averaged PM2.5. For 30-day averaged PM 2.5 and 60-day averaged PM2.5 we did not observe any statistically significant associations. We observed an increase in lymphocyte proliferation index stimulated by the plant protein phytohemagglutinin (PHA) at 1 of 2 PHA concentrations in association with modeled NO2. For the three inflammatory markers, we observed no notable associations with any of our measures of air pollution. If confirmed, our

  7. Influence of ventilation regimen on micro-environment and on ewe welfare and milk yield in summer

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    Mariangela Caroprese

    2010-01-01

    Full Text Available The effects of ventilation regimen on air quality, and on the welfare and production performance of thirty-six Comisanaewes were assessed in a 6-week trial conducted during the summer of 2002. Animals were divided into three groups of12, and subjected to the following treatments: low ventilation regimen providing a mean ventilation rate (VR of 35 m3/hper ewe, split in 30 min ventilation cycles at an air speed of 2 m/s (LOV-30; moderate ventilation regimen (VR = 70m3/h per ewe split in 30 min ventilation cycles at an air speed of 4 m/s (MOV-30; moderate ventilation regimen (VR =70 m3/h per ewe split in 60 min ventilation cycles at an air speed of 2 m/s (MOV-60. Air concentrations of microorganisms,dust, and gaseous pollutants were measured twice weekly. Respiration rate (RR and rectal temperature (RTwere monitored throughout the trial at 0830 and at 1400. Behavioral traits of ewes were recorded twice per week from0900 to 1200 and from 1500 to 1800. Cell-mediated immune response to phytohemagglutinin (PHA and humoralimmune response to chicken egg albumin were determined. At d 37 ewes were injected with porcine ACTH, and subjectedto blood sampling for evaluation of cortisol concentrations immediately before and 1, 2 and 4 h after ACTH injection.Milk yield was recorded daily. Individual milk samples were analyzed for composition, renneting parameters, somaticcell count (SCC, and bacteriological characteristics. Averages of maximum THI were about 3 points higher in the LOV-30 and the MOV-30 than in the MOV-60 room, whereas no differences emerged in the air concentrations of dust, gaseouspollutants and microorganisms. Significant interactions of treatment x time (P and for the time the ewes spent lying, idling and eating in the afternoon during weeks 2 and 3 of the study period.Significant effects of ventilation regimen x time (P the LOV-30 ewes giving smaller volumes of milk with a deteriorated coagulating behavior than those of the MOV-60 group

  8. Analysis of lead toxicity in human cells

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    Gillis Bruce S

    2012-07-01

    Full Text Available Abstract Background Lead is a metal with many recognized adverse health side effects, and yet the molecular processes underlying lead toxicity are still poorly understood. Quantifying the injurious effects of lead is also difficult because of the diagnostic limitations that exist when analyzing human blood and urine specimens for lead toxicity. Results We analyzed the deleterious impact of lead on human cells by measuring its effects on cytokine production and gene expression in peripheral blood mononuclear cells. Lead activates the secretion of the chemokine IL-8 and impacts mitogen-dependent activation by increasing the secretion of the proinflammatory cytokines IL-6 and TNF-α and of the chemokines IL-8 and MIP1-α in the presence of phytohemagglutinin. The recorded changes in gene expression affected major cellular functions, including metallothionein expression, and the expression of cellular metabolic enzymes and protein kinase activity. The expression of 31 genes remained elevated after the removal of lead from the testing medium thereby allowing for the measurement of adverse health effects of lead poisoning. These included thirteen metallothionein transcripts, three endothelial receptor B transcripts and a number of transcripts which encode cellular metabolic enzymes. Cellular responses to lead correlated with blood lead levels and were significantly altered in individuals with higher lead content resultantly affecting the nervous system, the negative regulation of transcription and the induction of apoptosis. In addition, we identified changes in gene expression in individuals with elevated zinc protoporphyrin blood levels and found that genes regulating the transmission of nerve impulses were affected in these individuals. The affected pathways were G-protein mediated signaling, gap junction signaling, synaptic long-term potentiation, neuropathic pain signaling as well as CREB signaling in neurons. Cellular responses to lead were

  9. Expression profiles of the immune genes CD4, CD8β, IFNγ, IL-4, IL-6 and IL-10 in mitogen-stimulated koala lymphocytes (Phascolarctos cinereus by qRT-PCR

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    Iona E. Maher

    2014-03-01

    Full Text Available Investigation of the immune response of the koala (Phascolarctos cinereus is needed urgently, but has been limited by scarcity of species-specific reagents and methods for this unique and divergent marsupial. Infectious disease is an important threat to wild populations of koalas; the most widespread and important of these is Chlamydial disease, caused by Chlamydia pecorum and Chlamydia pneumoniae. In addition, koala retrovirus (KoRV, which is of 100% prevalence in northern Australia, has been proposed as an important agent of immune suppression that could explain the koala’s susceptibility to disease. The correct balance of T regulatory, T helper 1 (Th1 and Th2 lymphocyte responses are important to an individual’s susceptibility or resistance to chlamydial infection. The ability to study chlamydial or KoRV pathogenesis, effects of environmental stressors on immunity, and the response of koalas to vaccines under development, by examining the koala’s adaptive response to natural infection or in-vitro stimulation, has been limited to date by a paucity of species- specific reagents. In this study we have used cytokine sequences from four marsupial genomes to identify mRNA sequences for key T regulatory, Th1 and Th2 cytokines interleukin 4 (IL-4, interleukin 6 (IL-6, interleukin 10 (IL-10 and interferon gamma (IFNγ along with CD4 and CD8β. The koala sequences used for primer design showed >58% homology with grey short-tailed opossum, >71% with tammar wallaby and 78% with Tasmanian devil amino acid sequences. We report the development of real-time RT-PCR assays to measure the expression of these genes in unstimulated cells and after three common mitogen stimulation protocols (phorbol myristate acetate/ionomycin, phorbol myristate acetate/phytohemagglutinin and concanavalin A. Phorbol myristate acetate/ionomycin was found to be the most effective mitogen to up-regulate the production of IL-4, IL-10 and IFNγ. IL-6 production was not

  10. Human embryonic stem cell-derived mesenchymal stroma cells (hES-MSCs engraft in vivo and support hematopoiesis without suppressing immune function: implications for off-the shelf ES-MSC therapies.

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    Ou Li

    Full Text Available Mesenchymal stroma cells (MSCs have a high potential for novel cell therapy approaches in clinical transplantation. Commonly used bone marrow-derived MSCs (BM-MSCs, however, have a restricted proliferative capacity and cultures are difficult to standardize. Recently developed human embryonic stem cell-derived mesenchymal stroma cells (hES-MSCs might represent an alternative and unlimited source of hMSCs. We therefore compared human ES-cell-derived MSCs (hES-MP002.5 cells to normal human bone marrow-derived MSCs (BM-MSCs. hES-MP002.5 cells had lower yet reasonable CFU-F capacity compared with BM-MSC (8±3 versus 29±13 CFU-F per 100 cells. Both cell types showed similar immunophenotypic properties, i.e. cells were positive for CD105, CD73, CD166, HLA-ABC, CD44, CD146, CD90, and negative for CD45, CD34, CD14, CD31, CD117, CD19, CD 271, SSEA-4 and HLA-DR. hES-MP002.5 cells, like BM-MSCs, could be differentiated into adipocytes, osteoblasts and chondrocytes in vitro. Neither hES-MP002.5 cells nor BM-MSCs homed to the bone marrow of immune-deficient NSG mice following intravenous transplantation, whereas intra-femoral transplantation into NSG mice resulted in engraftment for both cell types. In vitro long-term culture-initiating cell assays and in vivo co-transplantation experiments with cord blood CD34+ hematopoietic cells demonstrated furthermore that hES-MP002.5 cells, like BM-MSCs, possess potent stroma support function. In contrast to BM-MSCs, however, hES-MP002.5 cells showed no or only little activity in mixed lymphocyte cultures and phytohemagglutinin (PHA lymphocyte stimulation assays. In summary, ES-cell derived MSCs might be an attractive unlimited source for stroma transplantation approaches without suppressing immune function.

  11. A chromosomal breakage syndrome with profound immunodeficiency.

    Science.gov (United States)

    Conley, M E; Spinner, N B; Emanuel, B S; Nowell, P C; Nichols, W W

    1986-05-01

    The chromosomal breakage syndromes--ataxia-telangiectasia, Fanconi's anemia, and Bloom's syndrome--are associated with growth failure, neurologic abnormalities, immunodeficiency, and an increased incidence of malignancy. The relationship between these features is unknown. We recently evaluated a 21-year-old female with more severe chromosomal breakage, immunodeficiency, and growth failure than in any of the mentioned disorders. As of November 1985, the patient remains clinically free of malignancy. At age 18, the patient's weight was 22.6 kg (50th percentile for seven years), height was 129 cm (50th percentile for eight years), and head circumference was 42 cm (50th percentile for six months). Laboratory studies demonstrated a marked decrease in both B and T cell number and function. The peripheral blood contained 400 to 900 lymphocytes/microL with 32% T11 cells, 17% T4 cells, and 21% T8 cells. The proliferative responses to phytohemagglutinin (PHA), pokeweed mitogen, and concanavalin A were less than 10% of control. There were 1% surface IgM positive cells, and serum IgG was 185 mg/dL, IgM 7 mg/dL, IgA 5 mg/dL. In lymphocyte cultures stimulated with the T cell mitogens PHA, phorbol ester, and interleukin 2, 55% of the banded metaphases demonstrated breaks or rearrangements. The majority of the breaks involved four fragile sites on chromosomes 7 and 14, 7p13, 7q35, 14q11, and 14q32. These are the sites of the genes for the T cell-antigen receptor and the immunoglobulin heavy chain and are sites of gene rearrangement in lymphocyte differentiation. Epstein-Barr virus stimulated B cells and fibroblast cultures also demonstrated a high incidence of breaks, but the sites were less selective. These findings suggest that the sites of chromosomal fragility in the chromosomal breakage syndromes may be informative and that factors other than the severity of the immunodeficiency or the high incidence of chromosomal damage may contribute to the occurrence of malignancy in the

  12. Effect of Ciprofloxacin on the Immune Response of Growing Piglets to Streptococcosis suis

    Institute of Scientific and Technical Information of China (English)

    QING Liu-ting; YUAN Zong-hui

    2003-01-01

    24 seven-week-old hybrid healthy pigs (weighted 25.5?.4 kg ) from a source were randomly allotted to four groups (six each): the blank group, the negative group, the positive group and the test group. Pigs in the negative group and the test group were developed into a typical subacute Streptococcosis suis by inoculating subcutaneously 0.45 billion of pure living Streptococcus suis (type C55126) per kilogram body weight. Pigs in the positive group and the test group were intravenously injected ciprofloxacin (5 mg kg1 of BW) for 8 consecutive days (twice daily). Pigs in the blank group and positive group were inoculated with a placebo (0.85% NaCl). Some immunity parameters were tested on day 0, 7, 14, 21 and 28 after inoculation. Total leukocytes were counted with microscope. Differential leukocyte counts were performed on Giemsa-stained blood smears. The effect on the nitroblue tetrazolium (NBT) reduction of neutrophil was used to determine its phagocytic ability. Lymphocyte proliferation was determined by using the mitogens PHA. The total serum IgG concentration was measured by radial single immunodiffusion. A 50% hemolytic test was used to investigate the complement activity in serum. Whereas, a phytohemagglutinin (PHA) skin test was used to investigate in vivo immunity. We got the following results: Ciprofloxacin can retard the clinical signs of an increase of leukocyte counts, a large percent of lymphocytes and a relative percentage decrease of neutrophis. Ciprofloxacin could promote nitroblue tetrazolium (NBT) reduction by neutrophil. Though there was no obvious difference (P>0.05) between the test group and the negative group, the interaction of both Streptococcosis suis and ciprofloxacin was significantly different(P0.05) between the test group and the blank group. Delayed-type hypersensitivity (DTH) to PHA was increased in diseased animals after receiving the treatment with ciprofloxacin. Owing to the difference of interaction (P<0.01, on day 21, 28 after

  13. Mesothelin-MUC16 binding is a high affinity, N-glycan dependent interaction that facilitates peritoneal metastasis of ovarian tumors

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    Sathyanarayana Bangalore K

    2006-10-01

    Full Text Available Abstract Background The mucin MUC16 and the glycosylphosphatidylinositol anchored glycoprotein mesothelin likely facilitate the peritoneal metastasis of ovarian tumors. The biochemical basis and the kinetics of the binding between these two glycoproteins are not clearly understood. Here we have addressed this deficit and provide further evidence supporting the role of the MUC16-mesothelin interaction in facilitating cell-cell binding under conditions that mimic the peritoneal environment. Results In this study we utilize recombinant-Fc tagged human mesothelin to measure the binding kinetics of this glycoprotein to MUC16 expressed on the ovarian tumor cell line OVCAR-3. OVCAR-3 derived sublines that did not express MUC16 showed no affinity for mesothelin. In a flow cytometry-based assay mesothelin binds with very high affinity to the MUC16 on the OVCAR-3 cells with an apparent Kd of 5–10 nM. Maximum interaction occurs within 5 mins of incubation of the recombinant mesothelin with the OVCAR-3 cells and significant binding is observed even after 10 sec. A five-fold molar excess of soluble MUC16 was unable to completely inhibit the binding of mesothelin to the OVCAR-3 cells. Oxidation of the MUC16 glycans, removal of its N-linked oligosaccharides, and treatment of the mucin with wheat germ agglutinin and erythroagglutinating phytohemagglutinin abrogates its binding to mesothelin. These observations suggest that at least a subset of the MUC16-asscociated N-glycans is required for binding to mesothelin. We also demonstrate that MUC16 positive ovarian tumor cells exhibit increased adherence to A431 cells transfected with mesothelin (A431-Meso+. Only minimal adhesion is observed between MUC16 knockdown cells and A431-Meso+ cells. The binding between the MUC16 expressing ovarian tumor cells and the A431-Meso+ cells occurs even in the presence of ascites from patients with ovarian cancer. Conclusion The strong binding kinetics of the mesothelin-MUC16

  14. Immunomodulatory effects of oral antidiabetic drugs in lymphocyte cultures from patients with type 2 diabetes Efeito imunomodulador de hipoglicemiantes orais em cultura de linfócitos de pacientes com diabetes tipo 2

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    Karina Faccio Mello

    2011-02-01

    Full Text Available INTRODUCTION AND OBJECTIVE: It has been suggested that type 2 diabetes is an inflammatory response manifestation. The main drugs used to treat type 2 diabetes are sulphonylureas and biguanides. The aim of this study was to demonstrate the modulatory effects of oral hypoglycemic drugs (chlorpropamide and metformin on lymphocyte proliferation in vitro and ex vivo. METHODS: Peripheral blood mononuclear cells were isolated from human blood by gradient centrifugation. T-lymphocytes were stimulated by phytohemagglutinin (PHA and oral hypoglycemic drugs. RESULTS: In both in vitro and ex vivo experiments, there was a reduction in cell proliferation after treatment with oral hypoglycemic drugs. When both drugs were used in combination, a high level of cytotoxicity was observed, which made analysis of immunomodulatory effects unfeasible. DISCUSSION AND CONCLUSION: We demonstrated that diabetes itself may reduce cell proliferation significantly when stimulated by PHA, which may indicate that diabetic patients have difficulties in promoting an efficient inflammatory response. Moreover, the use of oral hypoglycemic drugs may aggravate this situation.INTRODUÇÃO E OBJETIVOS: Tem sido sugerido que o diabetes mellitus tipo 2 (DM2 é uma manifestação da resposta inflamatória. As principais drogas utilizadas no tratamento do DM2 são as sulfonilureias e as biguanidas. O objetivo deste trabalho é demonstrar os efeitos moduladores na proliferação de linfócitos causada pelos hipoglicemiantes orais (clorpropamida e metformina, in vitro e ex vivo. MÉTODOS: Células mononucleares de sangue periférico foram isoladas de seres humanos por gradiente de centrifugação. Os linfócitos T foram estimulados com fito-hemaglutinina (PHA e hipoglicemiantes. RESULTADOS: Nos experimentos in vitro e ex vivo, mostramos a redução da proliferação celular quando do tratamento com drogas hipoglicemiantes orais. Quando as drogas foram utilizadas em combinação, foi

  15. Linkage disequilibrium at the APA insecticidal seed protein locus of common bean (Phaseolus vulgaris L.).

    Science.gov (United States)

    Blair, Matthew W; Prieto, Sergio; Díaz, Lucy M; Buendía, Héctor F; Cardona, César

    2010-04-29

    An interesting seed protein family with a role in preventing insect herbivory is the multi-gene, APA family encoding the alpha-amylase inhibitor, phytohemagglutinin and arcelin proteins of common bean (Phaseolus vulgaris). Variability for this gene family exists and has been exploited to breed for insect resistance. For example, the arcelin locus has been successfully transferred from wild to cultivated common bean genotypes to provide resistance against the bruchid species Zabrotes subfasciatus although the process has been hampered by a lack of genetic tools for and understanding about the locus. In this study, we analyzed linkage disequilibrium (LD) between microsatellite markers at the APA locus and bruchid resistance in a germplasm survey of 105 resistant and susceptible genotypes and compared this with LD in other parts of the genome. Microsatellite allele diversity was found to vary with each of the eight APA-linked markers analyzed, and two markers within the APA locus were found to be diagnostic for bruchid resistance or susceptibility and for the different arcelin alleles inherited from the wild accessions. Arc1 was found to provide higher levels of resistance than Arc5 and the markers in the APA locus were highly associated with resistance showing that introgression of this gene-family from wild beans provides resistance in cultivated beans. LD around the APA locus was found to be intermediate compared to other regions of the genome and the highest LD was found within the APA locus itself for example between the markers PV-atct001 and PV-ag004. We found the APA locus to be an important genetic determinant of bruchid resistance and also found that LD existed mostly within the APA locus but not beyond it. Moderate LD was also found for some other regions of the genome perhaps related to domestication genes. The LD pattern may reflect the introgression of arcelin from the wild into the cultivated background through breeding. LD and association studies for

  16. Linkage disequilibrium at the APA insecticidal seed protein locus of common bean (Phaseolus vulgaris L.

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    Buendía Héctor F

    2010-04-01

    Full Text Available Abstract Background An interesting seed protein family with a role in preventing insect herbivory is the multi-gene, APA family encoding the α-amylase inhibitor, phytohemagglutinin and arcelin proteins of common bean (Phaseolus vulgaris. Variability for this gene family exists and has been exploited to breed for insect resistance. For example, the arcelin locus has been successfully transferred from wild to cultivated common bean genotypes to provide resistance against the bruchid species Zabrotes subfasciatus although the process has been hampered by a lack of genetic tools for and understanding about the locus. In this study, we analyzed linkage disequilibrium (LD between microsatellite markers at the APA locus and bruchid resistance in a germplasm survey of 105 resistant and susceptible genotypes and compared this with LD in other parts of the genome. Results Microsatellite allele diversity was found to vary with each of the eight APA-linked markers analyzed, and two markers within the APA locus were found to be diagnostic for bruchid resistance or susceptibility and for the different arcelin alleles inherited from the wild accessions. Arc1 was found to provide higher levels of resistance than Arc5 and the markers in the APA locus were highly associated with resistance showing that introgression of this gene-family from wild beans provides resistance in cultivated beans. LD around the APA locus was found to be intermediate compared to other regions of the genome and the highest LD was found within the APA locus itself for example between the markers PV-atct001 and PV-ag004. Conclusions We found the APA locus to be an important genetic determinant of bruchid resistance and also found that LD existed mostly within the APA locus but not beyond it. Moderate LD was also found for some other regions of the genome perhaps related to domestication genes. The LD pattern may reflect the introgression of arcelin from the wild into the cultivated

  17. Effect of oral administration of Propionibacterium acnes on growth performance, DTH response and anti-OVA titers in goat kids

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    Luis Miguel Ferrer

    2013-01-01

    Full Text Available Immunostimulants are susbstances that stimuli the response of effector cells to activate the immune response such as antigen uptake, cytokine release or antibody response. These substances can increase resistence to infection by different types of microorganisms, reducing dependence of antibiotics used in livestock animals. Recent reports have demonstrated the positive effect of Propionibacterium acnes (P. acnes to control animal diseases. In this study, we evaluated the effect of the non-specific immunostimulant P. acnes on immunological functions and growth performance in goat kids. Twenty five goat kids served as control group (A and another 25 animals received P. acnes being the experimental group (B. Kids were challenged with ovalbumin (OVA to assess humoral immunity. To assess in vivo cell immunity, delayed type hypersensitivity (DTH test with phytohemagglutinin (PHA was used, clinical signs and body weight were recorded each week until 9 weeks of age when the experiment ended. Blood samples were obtained to analyze serum proteins fractions and anti-OVA specific antibodies. No clinical signs of disease and no differences (p>0.05 on body weight between groups were recorded (7.32±0.81 kg in group A, 7.13±0.65 kg in group B. Goat kids from group B had more total protein (59.8±5g/l and albumin levels (32.8±3.3g/l than goat kids from group A (56.6±5.7 g/l, 29.6±3.9 g/l respectively (p<0.05. DTH response in goat kids from group B on day 42 was higher (p<0.05 than group A. At day 63, goat kids from group receiving P. acnes had higher percentage (85.4 of anti-OVA IgM titers (p<0.05 than control group (57.7. In conclusion, the results showed that oral administration of P. acnes to goat kids improved some aspects of the immune system of the animals and it could be used to control goat diseases.

  18. Genomic analysis of storage protein deficiency in common bean (Phaseolus vulgaris

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    Sudhakar ePandurangan

    2016-03-01

    Full Text Available A series of genetically related lines of common bean (Phaseolus vulgaris L. integrate a progressive deficiency in major storage proteins, the 7S globulin phaseolin and lectins. SARC1 integrates a lectin-like protein, arcelin-1 from a wild common bean accession. SMARC1N-PN1 is deficient in major lectins, including erythroagglutinating phytohemagglutinin (PHA-E but not α-amylase inhibitor, and incorporates also a deficiency in phaseolin. SMARC1-PN1 is intermediate and shares the phaseolin deficiency. Sanilac is the parental background. To understand the genomic basis for variations in protein profiles previously determined by proteomics, the genotypes were submitted to short-fragment genome sequencing using an Illumina HiSeq 2000/2500 platform. Reads were aligned to reference sequences and subjected to de novo assembly. The results of the analyses identified polymorphisms responsible for the lack of specific storage proteins, as well as those associated with large differences in storage protein expression. SMARC1N-PN1 lacks the lectin genes pha-E and lec4-B17, and has the pseudogene pdlec1 in place of the functional pha-L gene. While the α-phaseolin gene appears absent, an approximately 20-fold decrease in β-phaseolin accumulation is associated with a single nucleotide polymorphism converting a G-box to an ACGT motif in the proximal promoter. Among residual lectins compensating for storage protein deficiency, mannose lectin FRIL and α-amylase inhibitor 1 genes are uniquely present in SMARC1N-PN1. An approximately 50-fold increase in α-amylase inhibitor like protein accumulation is associated with multiple polymorphisms introducing up to eight potential positive cis-regulatory elements in the proximal promoter specific to SMARC1N-PN1. An approximately 7-fold increase in accumulation of 11S globulin legumin is not associated with variation in proximal promoter sequence, suggesting that the identity of individual proteins involved in proteome

  19. Demonstration of interleukin 1 activity in apparently homogeneous specimens of the pI 5 form of rabbit endogenous pyrogen.

    Science.gov (United States)

    Hanson, D F; Murphy, P A

    1984-08-01

    Rabbit mononuclear cells from oil-induced peritoneal exudates were purified by centrifugation on Percoll gradients, suspended in tissue culture medium, and stimulated with opsonized Staphylococcus epidermidis. The supernatants from these macrophages caused fever when injected intravenously into rabbits (endogenous pyrogen [EP] activity). The EP activity was contained in two protein fractions, with pIs of 7.3 and ca. 5.0. The same fractions caused mouse thymocytes to incorporate tritiated thymidine when incubated in vitro with small quantities of phytohemagglutinin (interleukin 1 [IL-1] activity). The pI 5.0 form of EP was purified to apparent homogeneity by sequential use of ammonium sulfate precipitation, gel filtration, ion-exchange chromatography, hydrophobic chromatography, and high-resolution isoelectric focusing. EP and IL-1 activities were not separable by any of these procedures. Active fractions from isoelectric focusing were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Only one band was visible as judged by a silver staining method, and IL-1 activity could be recovered by renaturing eluates from the same region of sodium dodecyl sulfate gels run in parallel. An estimate of specific activity was made by comparing the intensity of stained bands of EP with the intensity of bands containing known quantities of lysozyme or RNase. By this criterion, the specific activity of purified pI 5 EP was between 17,000 and 58,000 degrees C U/mg of protein, and the specific activity in terms of IL-1 was between 59 million and 360 million U per mg of protein. These observations suggest that both EP and IL-1 activities can be expressed by a single molecular species. The implications of this coincidence are discussed. It was also shown that highly purified pI 5 EP obtained from macrophages stimulated in the presence of 14C-labeled amino acids contained significant 14C radioactivity. This suggests that the pI 5.0 EP, like the pI 7

  20. 应用Ⅱ型胶原为载体的异种异体移植修复关节软骨缺损%Type Ⅱ collagen as the carrier for xenogeneic chondrocyte transplantation for joint cartilage defect repair

    Institute of Scientific and Technical Information of China (English)

    沈雁; 唐毅; 钟灿灿; 梁佩红; 黄雪芳; 邹海燕; 陈鸿辉; 梁伟国

    2005-01-01

    背景:已有用Ⅱ型胶原作为载体的软骨细胞移植的实验动物模型,Ⅱ型胶原是否引起实验性的关节炎或者发生由Ⅱ型胶原介导的细胞毒的反应尚未明确.目的:检测用猪Ⅱ型胶原免疫新西兰大白兔的细胞免疫状态.设计:以实验动物为研究对象,观察对比的探索性研究.单位:一所市级医院创伤外科研究所.材料:实验于1999-08/2000-02在广州红十字会医院创伤外科研究所完成.新西兰大白兔6只,雌雄不限,体质量2.0~3.0kg.方法:用Ⅱ型胶原免疫新西兰大白兔60 d,定期抽取血浆检测抗Ⅱ型胶原抗体;第60天取兔的外周血淋巴细胞,取兔脾细胞、淋巴结分离淋巴细胞,进行体外2次Ⅱ型胶原刺激,检测由此引起的反应性的细胞增殖规律.随机分为两组,第1组加入不同浓度植物血凝素(phytohemagglutinin,PHA)作阳性对照,并测定非特异性免疫;第2组加入不同浓度Ⅱ型胶原,检测特异性免疫.主要观察指标:①兔抗Ⅱ型胶原抗体含量检测.②正常兔与免疫兔脾淋巴细胞增殖试验比较.③正常兔和免疫兔淋巴结淋巴细胞增殖试验比较.④正常兔和免疫兔外周血淋巴细胞增殖试验比较.结果:第21天抗体效价出现第1次高峰,再次注射抗原40 d后出现第2次高峰,维持20 d后逐渐下降,正常兔的淋巴细胞在PHA刺激下发生增殖,但对Ⅱ型胶原的第1次刺激不发生增殖,而免疫兔对PHA和Ⅱ型胶原的刺激均能发生显著的增殖,Ⅱ型胶原浓度为25 mg/L已有发生,在50mg/L出现高峰.结论:异种Ⅱ型胶原在一定浓度下,可以引起免疫兔的抗Ⅱ型胶原抗体的升高,并可引起兔脾、外周血淋巴细胞增殖,在体内引起细胞免疫反应,且可引起移植免疫性关节炎.%BACKGROUND: Type Ⅱ collagen has been used as the carrier for chondrocyte transplantation in animal models, but whether type Ⅱ collagen may cause arthritis or mediate cytotoxicity remains

  1. Effects of various inducers on the expression of P2X7 receptor in human peripheral blood mononuclear cells%不同诱导因子对人外周血单个核细胞P2X7受体表达的作用

    Institute of Scientific and Technical Information of China (English)

    张秀军; 郑国光; 马小彤; 林永敏; 宋玉华; 吴克复

    2005-01-01

    Regulation of P2X7 receptor expression isof interest because activation of this receptor by extracellular ATP triggers a wide variety of cell functions in leukocytes. However, its expression and modulation in human peripheral blood mononuclear cells (PBMC)and monocytes remain unclear. RT-PCR was used to detect the constitutive level of P2X7 receptor and the levels upon stimulation with bacteria, bacterial product, mitogen and various cytokines in human PBMC and monocytes. P2X7 receptor mRNA was detected in PBMC and monocytes. P2X7 receptor expression in PBMC was up-regulated by interleukin-2, -4, -6 (IL-2, IL-4, IL-6) tumour necrosis factor-α (TNF-α), lipopolysaccharide (LPS) and heat-inactivated Staphylococcus aureus Cowan strain Ⅰ (SAC). However,interferon-γ (IFN-γ), granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF)and phytohemagglutinin-M (PHA-M) had little effect on the expression of P2X7 receptor. Furthermore, LPS and M-CSF could upregulate P2X7 receptor expression in monocytes, while IFN-γ, TNF-α and GM-CSF had weak effects, but pretreatment with these inducers could not further enhance LPS-stimulated P2X7 receptor expression in monocytes. The results obtained demonstrate that inflammatory stimuli drive P2X7 expression, thus supporting the hypothesis that P2X7 receptor may play a role in the inflammatory responses against bacteria infection, which need further verification.%ATP激活P2X7受体可产生一系列的白细胞功能反应,因此P2X7受体的表达调控引起我们的兴趣.然而P2X7受体在正常人外周血单个核细胞(peripheral blood mononuclear cells,PBMC)、单核细胞中的表达调控机制尚未阐明.本文用半定量RT-PCR方法检测多种细胞因子、细菌抗原、丝裂原对P2X7受体表达的诱导作用,探索P2X7受体的诱导表达模式.结果表明,单个核细胞和单核细胞可检出P2X7受体的表达;白细胞介素2、4、6(interleukin-2、-4、-6

  2. Cell and humoral immunity in endemic pemphigus foliaceus Imunidade humoral e celular no pênfigo foliáceo endêmico

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    Silvia Regina C. Sartori Barravieira

    1995-02-01

    Full Text Available A study was conducted on 16 patients with pemphigus foliaceus, ten of them with the localized form (group G1 and six with the disseminated form (group G2. These patients were submitted to full blood counts, quantitation of mononuclear cell subpopulations by monoclonal antibodies, study of blastic lymphocyte transformation, and quantitation of circulating antibodies by the indirect immunofluorescence test, in order to correlate their clinical signs and symptoms and laboratory data with their immunological profile, and to determine the relationship between circulating autoantibody titers and lesion intensity and course of lesions under treatment. Leucocytosis was observed especially in group G2. All patients showed decreased relative CD3+ and CD4+ values and a tendency to decreased relative values of the CD8+ subpopulation. Blastic lymphocyte transformation indices in the presence of phytohemagglutinin were higher in patients (group G1+G2 than in controls. The indirect immunofluorescence test was positive in 100% of G2 patients and in 80% of G1 patients. The median value for the titers was higher in group G2 than in group G1. Analysis of the results as a whole permits us to conclude that cell immunity was preserved and that there was a relationship between antibody titers detected by the direct immunofluorescence test and extent of skin lesions.Foram avaliados dezesseis doentes portadores de pênfigo foliáceo endêmico, dez com a forma localizada da doença (Grupo G1 e seis com a forma disseminada (Grupo G2, com os objetivos de correlacionar o quadro clínico e laboratorial desses pacientes com o perfil imunológico dos mesmos, e verificar a relação dos títulos dos anticorpos antiepiderme circulantes, identificados pela imunofluorescência indireta, com intensidade da lesão e com a evolução das lesões em tratamento. Foram realizados: hemograma completo, quantificação de subpopulação de células mononucleares por anticorpos monoclonais

  3. Ouabain exacerbates activation-induced cell death in human peripheral blood lymphocytes

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    Mabel B. Esteves

    2005-06-01

    Full Text Available Lymphocytes activated by mitogenic lectins display changes in transmembrane potential, an elevation in the cytoplasmic Ca2+ concentrations, proliferation and/or activation induced cell death. Low concentrations of ouabain (an inhibitor of Na+,K+-ATPase suppress mitogen-induced proliferation and increases cell death. To understand the mechanisms involved, a number of parameters were analyzed using fluorescent probes and flow cytometry. The addition of 100nM ouabain to cultures of peripheral blood lymphocytes activated with 5µg/ml phytohemagglutinin (PHA did not modify the increased expression of the Fas receptor or its ligand FasL induced by the mitogen. However, treatment with ouabain potentiated apoptosis induced by an anti-Fas agonist antibody. A synergy between ouabain and PHA was also observed with regard to plasma membrane depolarization. PHA per se did not induce dissipation of mitochondrial membrane potential but when cells were also exposed to ouabain a marked depolarization could be observed, and this was a late event. It is possible that the inhibitory effect of ouabain on activated peripheral blood lymphocytes involves the potentiation of some of the steps of the apoptotic process and reflects an exacerbation of the mechanism of activation-induced cell death.Quando linfócitos são ativados por lectinas mitogênicas apresentam mudanças do potencial de membrana, elevação das concentrações citoplasmáticas de cálcio, proliferação e/ou morte celular induzida por ativação (AICD. Concentrações baixas de ouabaína (um inibidor da Na,K-ATPase suprimem a proliferação induzida por mitógenos e aumentam a morte celular. Para entender os mecanismos envolvidos, uma série de parâmetros foram avaliados usando sondas fluorescentes e citometria de fluxo. A adição de 100nM de ouabaína para culturas de linfócitos de sangue periférico ativadas por fitohemaglutinina (PHA não modificou o aumento de expressão do receptor Fas ou de

  4. Anti-inflammatory effect of parenteral fish oil lipid emulsion on human activated mononuclear leukocytes Efecto antiinflamatorio de la emulsión parenteral de lípidos con aceite de pescado en leucocitos mononucleares humanos activados

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    T. Manzoni Jacintho

    2009-06-01

    Full Text Available Background & aim: To compare the effect of fish oilbased (FO lipid emulsions (LE for parenteral administration with standard LE and a new FO containing LE composed of four different oils on the antigen presentation and inflammatory variables. Methods: Phytohemagglutinin (PHA activated human mononuclear leukocytes were cultured with different LE - Control: without LE; SO: soybean oil; SO/FO: soybean and FO (4:1; MCT/SO: medium chain triglycerides and SO (1:1; MCT/SO/FO: MCT/SO and FO (4:1 and SMOF: a new LE containing FO. Cytokine production was evaluated by ELISA, the expression of antigen-presenting and co-stimulatory surface molecules were analyzed by flow cytometry and lymphocyte proliferation was assessed by H³-Thymidine incorporation, after tetanus toxoid-induced activation. Results: All LE decreased the HLA-DR and increased CD28 and CD152 expression on monocytes/macrophages and lymphocytes surface (p Antecedentes & objetivo: Comparar el efecto de las emulsiones lipídicas (EL basadas en aceite de pescado (AP para la administración parenteral con las EL estándar y una nueva EL que contiene AP compuesta por cuatro aceites distintos sobre la presentación antigénica y las variables inflamatorias. Métodos: se cultivaron leucocitos mononucleares activados con fitohemaglutinina (PHA con diferentes EL - Control: sin EL; AS: aceite de soja; AS/AP: soja y AP (4:1; TCM/AS: triglicéridos de cadena media y AS (1:1; TCM/AS/AP: TCM/AS y AP (4:1 y SMOF: una nueva EL que contiene AP. Se evaluó la producción de citocinas mediante ELISA, se analizó la expresión de moléculas de superficie de presentación de antígeno y co-estimuladoras mediante citometría de flujo y se evaluó la proliferación linfocitaria mediante la incorporación de timidina-H³ tras la activación inducida por el toxoide tetánico. Resultados: Todas las EL disminuyeron la expresión de HLA-DR y aumentaron la expresión de CD28 y CD152 sobre superficie de monocitos

  5. Aquatic pollution-induced immunotoxicity in wildlife species.

    Science.gov (United States)

    Luebke, R W; Hodson, P V; Faisal, M; Ross, P S; Grasman, K A; Zelikoff, J

    1997-05-01

    determined that rats fed freeze-dried Baltic Sea herring had higher virus titers after challenge with rat cytomegalovirus (RCMV) than rats fed Atlantic Ocean herring; perinatal exposure of rats to oil extracted from Baltic herring also reduced the response to challenge with RCMV. Keith Grassman reported an association between exposure to polyhalogenated aryl hydrocarbons and decreased T cell immunity in the offspring of fish-eating birds (herring gulls and Capsian terns) at highly contaminated sites in the Great Lakes. The greatest suppression of skin test responses to phytohemagglutinin injection (an indicator of T cell immunity) was consistently found at sites with the highest contaminant concentrations. Judith Zelikoff addressed the applicability of immunotoxicity studies developed in laboratory-reared fish for detecting altered immune function in wild populations. She presented data from studies done in her laboratory with environmentally relevant concentrations of metals as examples. Although the necessity of proceeding with caution when extrapolating across species was emphasized, she concluded that published data, and results presented by the other Symposium participants, demonstrate that assays similar to those developed for use in laboratory rodents may be useful for detecting immune system defects in wildlife species directly exposed to toxicants present in the environment.

  6. Early effect of maternal allergic asthma on T-regulatory cells immune response in cord blood of offsprings%母亲过敏性哮喘病史对新生儿调节性T细胞的影响

    Institute of Scientific and Technical Information of China (English)

    刘晶; 张捷; 徐伟; 许溟宇; 任锦; 闫冰迪; 李成玉; 马忠森

    2012-01-01

    Objective: Examined the impairment of regulatory T cells in cord blood from offspring of allergic asthmatic mothers. Methods: Cord blood mononuclear cells from 62 healthy neonates ( 40 healthy mothers and 22 allergic asthmatic mothers ) were isolated , and cultured with stimuli: lipid A ( TLR4 ligands ), peptidoglycan ( Ppg-TLR2 ligands ), mitogen ( PHA, phytohemagglutinin ) and Dermatophagoides pteronyssinusl. And then the amount of CD4 + CD25 + Foxp3 + T cells was acounted with flow cytometry; cytokine concentrations were measured in supernatants by LUMINEX technology; the suppressive function of regulatory T cells were examinated by isolating and culturing of CD4 + CD25 + T cells and CD4 + CD25 ~ T cells in vitro. Results: Cord blood from offspring of allergic asthmatic mothers showed Ppg-induced fewer regulatory T cells ( CD4 + CD25 + Foxp3 + T,P =0. 03 ) and lower IL-10 secretion ( P =0. 03 ). Furthermore, the suppressive capacity of regulatory T cells was impaired in PHA-induced division and proliferation of T effector cells in cord blood of offspring from allergic asthmatic mothers ( P =0.05 ). Meanwhile, the suppressive capacity of regulatory T cells to IL-13 production by effector cells was partially impared ( P = 0.07 ). Conclusion: In offspring of allergic asthmatic mothers, regulatory T cells amount, and suppressive function were impaired at birth, which maybe potentially contribute to the the susceptibility to allergic diseases.%目的:研究母亲过敏性哮喘病史对新生儿调节性T细胞的影响.方法:收集62例胎儿脐带血[40例健康母亲(对照组),22例母亲有过敏性哮喘病史(哮喘组)],分离单个核细胞进行体外培养,每例样品均分别给予以下4种刺激:类脂A(LpA)-Toll样受体4的配体,肽多糖(Ppg)-Toll样受体2的配体,植物血凝素(PHA),屋尘螨提取物.培养3天后,应用流式细胞技术检测CD4+CD25+Foxp3+调节性T细胞数量;Luminex流式荧光仪检测特异性细胞因

  7. Role of cytokines in promoting immune escape of FasL-expressing human colon cancer cells

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    Tong Xu; Bao-Cun Sun; Qiang Li; Xi-Shan Hao

    2005-01-01

    AIM: To investigate the potential role of cytokines in promoting Fas ligand (FasL)-expressing colon cancer cells.METHODS: Immunohistochemical SABC method was used to observe the expression of Fas receptor and ligand in SW620 colon cancer cell line and Jurkat T cells in order to provide the morphological evidence for the functions of Fas receptor and ligand. To examine the cytotoxicity of effector cells, CytoTox96(R) non-radioactive cytotoxicity assay was adopted to measure the lactate dehydrogenase-releasing value after SW620 cells were co-cultured with Jurkat T lymphocytes.RESULTS: The FasL of colon cancer SW620 cells was positive. The positive substances were distributed in the cell membrane and cytoplasm. The Fas receptor of colon cancer SW620 cells was negative. The Fas receptor and ligand of Jurkat T lymphocytes tumed out to be positive. The positive substances were distributed in the cell membrane.After phytohemagglutinin (PHA)-stimulated Jurkat T lymphocytes were co-cultured with phorbol 12-myristate 13-acetate (PMA)-plus-ionomycin-stimulated (for 48 h) SW620ceils or tumor necrosis factor-alpha (TNF-α)-stimulated (for 48 h) SW620 cells or unstimulated SW620 cells for 4 h,the cytotoxicity of SW620 cells to PHA-stimulated Jurkat cells at effector-to-target ratios of 10:1, 5:1, 2.5:1, and 1.25:1was 74.6%, 40.8%, 32.4%, and 10.9% (F= 8.19, P<0.05);or 54.9%, 35.3%, 22.0%, and 10.3% (F= 11.12, P<0.05);or 14.9%, 10.5%, 6.9%, and 5.8% (F= 3.45, P<0.05).After PHA-stimulated Jurkat T lymphocytes were co-cultured with unstimulated SW620 cells for 8 h, the cytotoxicity of SW620 cells to PHA-stimulated Jurkat cells at effector-totarget ratios of 5:1, 2.5:1, and 1.25:1 from the experiment was 83.9%, 74.1%, and 28.5% (F= 137.04, P<0.05)respectively. Non-radioactive cytotoxicity assay showed that the apoptotic rate of Jurkat cells remarkably increased with the increase of planting concentration of SW620 cells and co-culture time after the SW620 cells were co

  8. The Immunoregulation effects of bone marrow mesenchymal stem cells on the lymphocytes of rats with type 1 diabetes mellitus in vitro%骨髓间充质干细胞体外对1型糖尿病大鼠淋巴细胞的免疫调节作用

    Institute of Scientific and Technical Information of China (English)

    李煜环; 宋振顺; 范子扬; 张福琴

    2011-01-01

    Objective To observe the effects of bone marrow mesenchymal stem cells (MSCs) on the lymphocytes of rats with type 1 diabetes mellitus (T1DM) in vitro, and investigate the inhibitory effect of MSCs on lymphocytes proliferation and the underlying mechanism. Methods MSCs were isolated from SD rats, cultured in vitro, purified and then identified by testing the phenotypes with flow cytometry (FCM). The third-generation MSCs were planted in 24-well plates. After treated with mitomycin C, MSCs were co-cultured for 72 h with the T1 DM rat's lymphocytes activated by phytohemagglutinin (PHA). The proliferation of lymphocyte was measured by methyl thiazol tetrazolium (MTT) method. FCM analysis was done to investigate the apoptosis, cell cycle and the proportion of CD4+ CD25+ regulatory T cells of the T1 DM rat's lymphocytes after co-cultivation. Results The phenotypes of MSCs from normal SD rats were CD29 + , CD90 +, CD106 + , CD34-, CD45 -. MSCs obviously inhibited the lymphocyte proliferation stimco-culture system, most of the lymphocytes were arrested at G0/G1 phase. The apoptosis rate of lymphocytes (58.05 ± 0. 89)% in group C was increased significantly as compared with the control group (43.35± 0.86 ) % ( P < 0. 05 ) as well as the proportion of CD4 + CD25 + regulatory T cells (22.76 ± 1.15 ) % vs (5.80 ± 0. 68) %. Conclusion In vitro, MSCs can obviously inhibit the T1 DM rat' s lymphocytes proliferation stimulated with PHA via increasing the proportion of CD4 + CD25 + regulatory T cells.%目的 观察骨髓间充质干细胞(MSCs)体外对1型糖尿病(T1DM)大鼠淋巴细胞表型及增殖能力的影响,探讨其抑制淋巴细胞增殖的机制.方法 分离、培养和鉴定大鼠MSCs,噻唑蓝(MIT)比色法观察该细胞对淋巴细胞增殖能力的影响,应用流式细胞术分析MSCs对植物血凝素(PHA)作用下淋巴细胞凋亡,周期水平和CD4+CD25+调节性T细胞亚群(CD4+CD25+Tregs)比例的影响.结果 大鼠MSCs表型为CD29+、CD90+

  9. N-acetylcysteine and fructose-1,6-bisphosphate: immunomodulatory effects on mononuclear cell culture N-acetilcisteína e frutose-1,6-bisfosfato: efeito imunomodulador em cultura de células mononucleares

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    Ricardo Obalski de Mello

    2012-04-01

    Full Text Available INTRODUCTION: Sepsis is a complex syndrome caused by an uncontrolled systemic inflammatory response. Inflammatory cytokines play a pivotal role in septic shock pathogenesis. Therapeutic strategies have been tested in order to modulate the excessive generation or function of sepsis mediators. OBJECTIVE: The objective of the present study was to investigate the therapeutic effect of N-acetylcysteine (NAC and its association with fructose-1,6-bisphosphate (FBP on T-lymphocytes proliferation, interleukin-1β (IL-1β and monocyte chemotactic protein-1 (MCP-1 levels. MATERIAL AND METHODS: Peripheral blood mononuclear cell samples were isolated from healthy individuals. T-lymphocytes were stimulated with phytohemagglutinin for 96 hours and submitted to different concentrations of NAC or NAC associated with FBP. RESULTS: NAC (10 and 15 mM and NAC (15 mM associated with FBP reduced T-lymphocytes proliferation. IL-1β levels rose in the presence of both NAC (15 mM and NAC with FBP (1.25 mM. MCP-1 levels were reduced only by NAC (15 mM associated with FBP (1.25 mM. CONCLUSION: The results suggest that both NAC itself and NAC associated with FBP inhibit cellular proliferation, acting as potent immunomodulatory agents, which corroborates its use in the treatment of inflammatory diseases.INTRODUÇÃO: A sepse é uma síndrome complexa causada pela resposta inflamatória sistêmica descontrolada. As citocinas inflamatórias representam papel central na patogênese do choque séptico. Têm sido testadas estratégias terapêuticas a fim de modular a geração ou a função excessiva de mediadores na sepse. OBJETIVO: O objetivo deste estudo foi investigar o efeito terapêutico da N-acetilcisteína (NAC e sua associação com a frutose-1,6-bisfosfato (FBP sobre a proliferação de linfócitos T e a geração de interleucina-1β (IL-1β e proteína quimiotática de monócitos 1 (MCP-1 em cultura celular. MATERIAL E MÉTODOS: Foram isoladas células mononucleares de

  10. In Ovo Vaccination with Turkey Herpesvirus Hastens Maturation of Chicken Embryo Immune Responses in Specific-Pathogen-Free Chickens.

    Science.gov (United States)

    Gimeno, Isabel M; Faiz, Nik M; Cortes, Aneg L; Barbosa, Taylor; Villalobos, Tarsicio; Pandiri, Arun R

    2015-09-01

    Administration of Marek's disease (MD) vaccines in ovo has become a common practice for the poultry industry. Efficacy of MD vaccines is very high, even though they are administered to chicken embryos that are immunologically immature. We have recently demonstrated that in ovo vaccination with turkey herpesvirus (HVT) results in increased activation of T cells at hatch. Our previous results suggested that in ovo vaccination with HVT might have a positive impact not only on MD protection but also on the overall maturity of the developing immune system of the chicken (Gallus gallus domesticus). The objective of this study was to evaluate the effect of administration of HVT at 18 days of embryonation (ED) on the maturation of the embryo immune system. Four experiments were conducted in Specific-Pathogen-Free Avian Supplies (SPAFAS) chickens to evaluate the effect of administration of HVT at 18 ED on the splenic cell phenotypes at day of age (experiment 1) and on the ability of 1-day-old chickens to respond to various antigens compared with older birds (experiments 2 and 3). In addition, a fourth experiment was conducted to elucidate whether administration of other serotype's MD vaccines (CVI988 and SB-1) at 18 ED had the same effect as HVT on the spleen cell phenotypes at day of age. Our results demonstrated that 1-day-old chickens that had received HVT in ovo (1-day HVT) had higher percentages of CD45+, MHC-I+, CD45+MHC-I+, CD3+, MHC-II+, CD3+MHC-II+, CD4+, CD8+, and CD4+CD8+ cells in the spleen than 1-day-old sham-inoculated chickens (1-day sham). Moreover, spleens of 1-day HVT chickens had greater percentages of CD45+MHC-I+ cells and equal or greater numbers of CD4+CD8- and CD4-CD8+ cells than older unvaccinated chickens. In addition, administration of HVT at 18 ED rendered chicks at hatch more responsive to unrelated antigens such as concavalin A, phytohemagglutinin-L, and keyhole limpet hemocyanin. Administration of MD vaccines of other serotypes had an effect

  11. TBX21 and HLX1 polymorphisms influence cytokine secretion at birth.

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    Vera Isabel Casaca

    Full Text Available BACKGROUND: TBX21 (T cell specific T-box transcription factor and HLX1 (H.20-like homeobox 1 are crucial transcription factors of T(H1-cells, inducing their differentiation and suppressing T(H2 commitment, particularly important for early life immune development. This study investigated the influence of TBX21 and HLX1 single nucleotide polymorphisms (SNPs, which have previously been shown to be associated with asthma, on T(H1/T(H2 lineage cytokines at birth. METHODS AND FINDINGS: Cord blood mononuclear cells (CBMCs of 200 neonates were genotyped for two TBX21 and three HLX1 SNPs. CBMCs were stimulated with innate (Lipid A, LpA; Peptidoglycan, Ppg, adaptive stimuli (house dust mite Dermatophagoides pteronyssinus 1, Derp1 or mitogen (phytohemagglutinin, PHA. Cytokines, T-cells and mRNA expression of T(H1/T(H2-related genes were assessed. Atopic diseases during the first 3 years of life were assessed by questionnaire answered by the parents. Carriers of TBX21 promoter SNP rs17250932 and HLX1 promoter SNP rs2738751 showed reduced or trendwise reduced (p≤0.07 IL-5, IL-13 and TNF-α secretion after LpA-stimulation. Carriers of HLX1 SNP rs2738751 had lower IL-13 levels following Ppg-stimulation (p = 0.08. Carriers of HLX1 exon 1 SNP rs12141189 showed increased IL-5 (LpA, p = 0.007; Ppg, p = 0.10, trendwise increased IL-13 (LpA, higher GM-CSF (LpA/Ppg, p≤0.05 and trendwise decreased IFN-γ secretion (Derp1+LpA-stimulation, p = 0.1. Homozygous carriers of HLX1 promoter SNP rs3806325 showed increased IL-13 and IL-6 (unstimulated, p≤0.03. In carriers of TBX21 intron 3 SNP rs11079788 no differences in cytokine secretion were observed. mRNA expression of T(H1/T(H2-related genes partly correlated with cytokines at protein level. TBX21 SNP rs11079788 carriers developed less symptoms of atopic dermatitis at 3 years of age (p = 0.03. CONCLUSIONS: Polymorphisms in TBX21 and HLX1 influenced primarily IL-5 and IL-13 secretion after Lp

  12. The diagnostic value of IFN-γ reaction of peripheral blood mononuclear cells induced by ESAT-6 in tuberculous meningitis%ESAT-6诱导的外周血单个核细胞γ-干扰素反应对结核性脑膜炎的诊断价值

    Institute of Scientific and Technical Information of China (English)

    程言博; 周延龙; 董艳; 李新华; 葛巍; 汪莉萍; 董瑞国

    2012-01-01

    Objective To investigate the diagnostic value of interferon-7 (INF-7) reaction of peripheral blood mononuclear cells (PBMC) induced by 6-kDa early secreted antigen target (ESAT-6) in tuberculous meningitis. Methods 62 cases of patients with meningitis and 20 cases of healthy control were recruited in this study. There were 20 cases of tuberculous meningitis (TBM group) , 25 cases of viral meningitis (VM group) and 17 cases of bacterial meningitis (BM group). PBMC was separated by Ficoll, and was cultured. Each group of PBMC was treated by EAST-6, purfied protein derivatives tuberculin (PPD), phytohemagglutinin (PHA) and normal saline(NS) respectively. The level of IFN-γ in culture supernatant was measured by ELBA kits . Results There were significant increase of IFN-γ in TBM group treated by ESAT-6 (compared to NS treatment, P <0.01). There were also increased reaction of IFN-7 in TBM group treated by PPD (compared to NS treatment, P<0.01) , however, the level was lower than ESAT-6 treatment (P<0.01) . By receiver operating characteristic (ROC) curve analysis , area under ROC curve of ESAT-6 treatment was 0.918 (P<0.01), 95% CI 0.831-1.0, and PPD treatment was 0.725 (P<0.05), 95% CI 0.56-0.89. Conclusion There are excellent sensitivity and specificity of ESAT-6 induced IFN-7 reaction of PBMC in diagnoseof tuberculous meningitis , and it is relatively more feasible in clinical practice.%目的 探讨6-kDa早期分泌靶抗原(ESAT-6)刺激的外周血单个核细胞(PBMC) γ-干扰素(IFN-γ)反应对结核性脑膜炎的诊断价值.方法 研究对象共82例,其中结核性脑膜炎组20例,病毒性脑膜炎组25例,一般细菌性脑膜炎组17例,并设健康对照组20例.Ficoll法分离PBMC并培养,分别以ESAT-6、纯化结核菌素(PPD)、植物血凝素(PHA)和生理盐水处理.ELISA法测定各组上清IFN-γ浓度.结果 ESAT-6处理引起结核性脑膜炎组显著的IFN-γ反应(与生理盐水处理相比P<0.01).PPD刺激在结核性脑膜炎

  13. 阿奇霉素对支气管哮喘患儿外周血辅助性T淋巴细胞9和辅助性T淋巴细胞17功能的影响%Influence of Azithromycin on peripheral blood helper T lymphocyte 9 cells and helper T lymphocyte 17 cells in ;children with bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    刘伟; 周娟; 孙莹; 张秋业

    2015-01-01

    Objective To observe the influence of Azithromycin on helper T lymphocyte 9 cells ( Th9 ) and Th17 in peripheral blood of children with bronchial asthma,and to investigate the immunomodulating effect of Azithro-mycin. Methods Twenty-six asthmatic children were selected as the experimental group,and 17 healthy children as a control group. Peripheral blood mononuclear cells(PBMC)were isolated from venous blood by density gradient cen-tri-fugation under aseptic conditions. PBMC were split by phytohemagglutinin( PHA) in vitro. Different concentrations of Azithromycin (0,0. 1,1. 0,10. 0 mg/L)were added into the cultures in the experimental group. The control group was not interfered with azithromycin. The supematant was collected after 72 h. The levels of interleukin ( IL)-4, IL-9,IL-17 and IL-23 in the supematant were determined by enzyme-linked immunosorbent assay(ELISA). SPSS 17. 0 software was used to analyze data. Results (1) The levels of IL-4,IL-9,IL-17 and IL-23 produced from PBMC of the experimental group were significantly higher than those in the control group(t=9. 210,3. 040,8. 965,2. 796,P0. 05). The levels of IL-17 and IL-23 of Azithromycin 10 mg/L were lower than those in Azithromycin 0 mg/L and 0. 1 mg/L(P0. 05). (3)IL-9 level was negatively correlated with IL-4 level in the experimental group(r=-0. 255,P>0. 05). The levels of IL-23 and IL-17 secreted by Th17 in asthmatic chil-dren had correlation. The secretion of IL-17 levels rose with the secretion of IL-23 rise(r=0. 64,P0.05)。阿奇霉素10.0 mg/L 组 IL-17、IL-23的分泌水平低于0 mg/L 组和0.1 mg/L 组(P0.05);3.哮喘组患儿PBMC经不同浓度阿奇霉素干预后分泌IL-9与IL-4水平间无相关性(r=-0.255,P>0.05),哮喘组PBMC分泌IL-17与IL-23水平呈正相关(r=0.64,P<0.05)。结论支气管哮喘患儿Th9和Th17细胞功能增强;阿奇霉素在较高组织浓度(10.0 mg/L)条件下可抑制Th9和 Th17细胞功能,前者与IL-4水平变化无关联,后者与IL-23水平降

  14. 喘可治对慢性阻塞性肺疾病患者外周血单个核细胞分泌白介素-17A的影响%Effects of Chuankezhi injection on interleukin-17A secretion of peripheral blood mononuclear cells in patients with chronic obstructive pulmonary disease

    Institute of Scientific and Technical Information of China (English)

    洪春霖; 洪敏俐; 陈慧暖; 李德秀; 宋小玲; 陈文喜; 高凌云; 柯庚申

    2013-01-01

    Objective To explore the effects of Chuankezhi (CKZ) injection on interleukin-17A (IL-17A) production of peripheral blood mononuclear cells (PBMCs) in patients with chronic obstructive pulmonary disease(COPD).Methods PBMCs from patients with stable COPD and health controls were cultured in RPML1640 media with phytohemagglutinin (PHA) plus different doses of CKZ (2%,4% and 6%).The concentration of IL 17A in the supernatants was determined by ELISA after 48 hours of incubation.Results Compared with baseline,the level of IL-17A stimulated by PHA was increased significantly in COPD group [(153.89±45.07)ng/L vs.(10.21±2.27)ng/L,t=12.265,P<0.001].The level of IL-17A stimulated by PHA was higher in COPD group than in health controls [(153.89 ± 45.07) ng/L vs.(110.23 ± 53.99) ng/L,t =2.404,P =0.023].The level of IL-17A was significantly decreased in COPD group after CKZ intervention,which showed a negative correlation with CKZ dose (r=-0.599,P<0.001).Conclusions CKZ can inhibit IL-17A production of PBMCs in patients with stable COPD,which may be one of the mechanisms of CKZ in COPD treatment.%目的 探讨喘可治注射液对慢性阻塞性肺疾病(COPD)稳定期患者外周血单个核细胞(PBMCs)分泌白介素-17A(IL-17A)的影响. 方法 分离稳定期COPD患者和健康对照的PBMCs,加入植物凝集素(PHA),并在PHA基础上加入不同浓度(2%、4%、6%)的喘可治,经培养48 h后,用酶联免疫吸附(ELISA)法测定培养物上清液中IL-17A浓度. 结果 经PHA诱导活化后,COPD组PBMCs培养物上清液IL-17A浓度明显升高,并高于健康对照组;COPD组活化后与活化前比较,[(153.89±45.07) ng/L比(10.21±2.27)ng/L;t=12.265,P<0.001],两组活化后比较[(153.89±45.07)ng/L比(110.23±53.99) ng/L;t=2.404,P=0.023];加入喘可治后,COPD组PBMCs培养物上清液IL-17A浓度降低,并呈剂量依赖负相关(r=-0.599,P<0.001). 结论 喘可治可以抑制COPD稳定期患者PBMCs分泌IL-17A,这可能是喘

  15. Estudo da proliferação linfocitária em pacientes sensibilizados ao níquel Study on lymphocyte proliferation in nickel sensitive patients

    Directory of Open Access Journals (Sweden)

    Ana Paula Galli Sanchez

    2005-04-01

    concentrations of Candida albicans antigen as well as pokeweed, phytohemagglutinin A and anti-CD3 antibody (OKT3 mitogens. Tritiated thymidine was added to plates, radioactivity incorporated by cells was measured and the results expressed by the stimulation index (SI. RESULTS: The lymphocyte proliferative response was higher in cases than in controls in all nickel concentrations tested. Considering positive test reactions when SI > 3, none of the controls and 16 (84.21% cases were positive in at least one of five concentrations used. The proliferative responses to Candida albicans and mitogens were similar in cases and controls, demonstrating normal cellular immunity in both groups. CONCLUSION: The lymphocyte proliferation test is useful in diagnosis of nickel sensitivity.

  16. CHANGING METABOLIC FUNCTIONS IN EXPERIMENTAL ANIMALS AFTER INTRODUCTION OF THE XENOBIOTIC, IMMUNOTROPIC DRUG AND PROBIOTIC

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    Zvyagintseva O.V.

    2015-05-01

    Saccharomyces cerevisiae. The peripheral blood leukocytes were cultured according to the method of Hereford in medium 199 with the addition of fetal calf serum in the absence and in the presence of T-cell mitogen – phytohemagglutinin. Results and discussion. In all studied groups (introduction of the xenobiotic, "Fungidol", probiotic experimental animals revealed a significant increase in the concentrations of ceruloplasmin and haptoglobin on the average in 1,5 times in comparison with the control, indicating the development of the inflammatory process after the toxic action of copper sulphate. During administration of sulphate of copper, the experimental animals showed a reduction in the index of completion of phagocytosis, indicating a failure of the process of endocytosis of bacterial antigens and reduced stimulation index due to the low activity of NADPoxidase system of phagocytes. The introduction of xenobiotic animals was increased 1,2 times compared with the control (23,33±1,38 % the number of transformed cells in the background of mitogenic inducer of cell proliferation. The proliferative activity of hemolytic after the joint action of the xenobiotic and immunotropic drug in cell culture with the mitogen was the highest and exceeded 1,5 times control (23,33±1,38%. After the introduction of copper sulfate and probiotic proliferative activity of hemolytic was also significantly higher spontaneous. Introduction biologic response modifier substance to a greater extent than probiotics stimulate a protective immune processes aimed at combating the negative effect of the xenobiotic. Conclusion. Thus, the introduction of copper sulfate launches in animals a cascade of reactions aimed at the disruption of homeostasis. It is a violation of various physiological processes of digestion, respiration, cell differentiation, water-salt metabolism, metabolism of carbohydrates, proteins, lipids, detoxification of exogenous substrates and metabolites, production of biologically active

  17. 复方甘草酸苷对斑秃患者外周血单一核细胞中y干扰素、肿瘤坏死因子β表达的调节作用%Regulatory effect of glycyrrhizin on the expression of interferon (IFN)-γ and tumor necrosis factor (TNF)-β in peripheral blood mononuclear cells of patients with alopecia areata

    Institute of Scientific and Technical Information of China (English)

    秦小卫; 陈丽芳

    2011-01-01

    目的 探讨甘草酸苷对斑秃患者外周血单一核细胞(PBMC)γ干扰素(IFN-γ)、肿瘤坏死因子β(TNF-β)表达的调节作用.方法 轻度斑秃患者18例、重症斑秃患者24例及正常人20例为检测对象,用密度梯度离心法常规分离PBMC,用逆转录-聚合酶链反应检测PBMC经植物血凝素和复方甘草酸苷共同刺激后IFN-γ和TNF-β的表达水平.结果 重症斑秃组患者IFN-γ及TNF-β水平均明显高于轻度斑秃组及正常人对照组(P值均< 0.05),轻度斑秃组高于正常人对照组(P值均<0.05).甘草酸苷与植物血凝素共同刺激斑秃患者PBMC后可下调IFN--γ及TNF-β表达水平(P值均<0.05).结论 甘草酸苷可抑制斑秃患者Th1型细胞因子表达,逆转Th1型反应.%Objective To observe the regulatory effect of glycyrrhizin on the expression of IFN-γ and TNF-β in peripheral blood mononuclear cells (PBMCs) of patients with alopecia areata.Methods PBMCs were obtained from 18 patients with mild alopecia areata,24 patients with severe alopecia areata and 20 normal human controls,and cultured with phytohemagglutinin (PHA) or the combination of PHA and glycyrrhizin for 24 hours.Then,reverse transcription (RT)-PCR was conducted to detect the mRNA expression of IFN-γand TNF-β in these cells.Results The mRNA expression levels of IFN-γand TNF-β in PBMCs were significantly higher in patients with severe alopecia areata than in those with mild alopecia areata and normal human controls (all P < 0.05),and higher in patients with mild alopecia areata than in normal human controls (both P < 0.05).A significant decrease was observed in the mRNA expressions of IFN-γ and TNF-β in the PBMCs from patients with alopecia areata after stimulation with the combination of PHA and glycyrrhizin (both P <0.05).Conclusion Glycyrrhizin can inhibit the expression of Th1-type cytokines and reverse Th1-type immune response.

  18. 罗格列酮上调成人隐匿性自身免疫糖尿病患者CD4+T细胞Foxp3 mRNA的表达%Rosiglitazone upregulates Foxp3 mRNA expression of CD4+T cells in adults with latent autoimmune diabetes

    Institute of Scientific and Technical Information of China (English)

    杨治芳; 周智广; 黄干; 彭健; 颜湘

    2008-01-01

    目的 观察罗格列酮对成人隐匿性自身免疫糖尿病(LADA)患者CD4+调节性T细胞的影响,旨在探讨罗格列酮的免疫调节机制.方法 采用磁珠分离LADA患者CD4+T细胞,1、10和100 μmol/L罗格列酮干预CIM+T细胞.MTT法检测细胞活性,3H-TdR掺人法检测增殖抑制率.流式细胞术检测CD4+CD25+T细胞比值.RT-PCR检测过氧化物酶体增殖激活受体-γ(PPARγ)mRNA、TGF-β1 mRNA表达,实时荧光定量PCR检测Foxp3 mRNA表达.结果 CD4+T细胞表达PPARγmRNA.罗格列酮抑制植物凝集素(PHA)刺激的CD4+T细胞增殖.1μmol/L和10μmol/L组罗格列酮作用的CD4+CD25+T细胞比例无明显变化,100 μmol/L组CD4+CD25+T细胞比例降低.10μmol/L罗格列酮上调CD4+T细胞的Foxp3 mRNA表达,但TGF-β1 mRNA表达无显著变化.小剂量IL-2参与下,1μmol/L罗格列酮(药理浓度范围内)升高CD4+T细胞Foxp3mRNA表达.结论 罗格列酮上调LADA患者CD4+T细胞Foxp3 mRNA表达,改善自身免疫耐受缺陷.%Objective To investigate the effect of rosiglitazone on the CD4+regulatory T cells in the patients with latent autoimmune diabetes in adults(LADA).Methods The CIM+T cells from IADA patients were isolated with anti-CD4-dynal magnetic beads.The expression of Foxp3 mRNA,along with peroxisome proliferators activator receptors gamma(PPARγ)mRNA and TGF-131 mRNA was determined.The effect of rosiglitazone on CD4+T cells was measured,after treated with rosiglitazone for 48 h.Cell viability was assessed by Mtit assay.The proliferation was assayed with 3 H-TdR.Two-color staining(anti-CD4,anti-CD25)flow cytometric analysis was employed to measure the percentage of CD4+CD25+T cells of Deriph eral blood.Resuits PPARγmRNA was expressed in peripheral CD4+T lymphocytes.RosiglitazoBe inhibited phytohemagglutinin(PHA)-induced human CD4+T cell proliferation in dose dependence.The percentage of CD4+CD25+T cells showed no significant change after the peripheral blood culture with 1 μmol/Land 10

  19. Multiparameter fluorescence imaging for quantification of TH-1 and TH-2 cytokines at the single-cell level

    Science.gov (United States)

    Fekkar, Hakim; Benbernou, N.; Esnault, S.; Shin, H. C.; Guenounou, Moncef

    1998-04-01

    Immune responses are strongly influenced by the cytokines following antigenic stimulation. Distinct cytokine-producing T cell subsets are well known to play a major role in immune responses and to be differentially regulated during immunological disorders, although the characterization and quantification of the TH-1/TH-2 cytokine pattern in T cells remained not clearly defined. Expression of cytokines by T lymphocytes is a highly balanced process, involving stimulatory and inhibitory intracellular signaling pathways. The aim of this study was (1) to quantify the cytokine expression in T cells at the single cell level using optical imaging, (2) and to analyze the influence of cyclic AMP- dependent signal transduction pathway in the balance between the TH-1 and TH-2 cytokine profile. We attempted to study several cytokines (IL-2, IFN-(gamma) , IL-4, IL-10 and IL-13) in peripheral blood mononuclear cells. Cells were prestimulated in vitro using phytohemagglutinin and phorbol ester for 36h, and then further cultured for 8h in the presence of monensin. Cells were permeabilized and then simple-, double- or triple-labeled with the corresponding specific fluorescent monoclonal antibodies. The cell phenotype was also determined by analyzing the expression of each of CD4, CD8, CD45RO and CD45RA with the cytokine expression. Conventional images of cells were recorded with a Peltier- cooled CCD camera (B/W C5985, Hamamatsu photonics) through an inverted microscope equipped with epi-fluorescence (Diaphot 300, Nikon). Images were digitalized using an acquisition video interface (Oculus TCX Coreco) in 762 by 570 pixels coded in 8 bits (256 gray levels), and analyzed thereafter in an IBM PC computer based on an intel pentium processor with an adequate software (Visilog 4, Noesis). The first image processing step is the extraction of cell areas using an edge detection and a binary thresholding method. In order to reduce the background noise of fluorescence, we performed an opening

  20. Crude propolis as an immunostimulating agent in broiler feed during the first stagePrópolis bruta como agente imunoestimulante na alimentação de frangos de corte na fase inicial

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    Cinthia Eyng

    2013-10-01

    Full Text Available The experiment herein was conducted to evaluate the efficacy of supplementing a broiler diet with crude propolis on the animals’ immune response (humeral and cellular, lymphoid organ weight and hematological profile. One hundred sixty-eight male broilers raised in metabolic cages until 21 days of age were used in the experiments. The birds were randomly distributed in an experimental design with six treatments that consisted of different crude propolis doses (0, 100, 200, 300, 400 and 500 ppm; the experimental unit was repeated seven times with four birds each. Including crude propolis in the food did not affect (P0.05. The interdigital reaction to the phytohemagglutinin displayed quadratic behavior as a function of time and crude propolis dose. Using the adjusted equation, the 275.45 ppm crude propolis dose and 39.35 hours produced the lowest and highest reaction values. Inclusion of 100 ppm crude propolis in the broiler feed was an effective immunostimulatory agent for cell-mediated responses. Este experimento foi realizado para avaliar a eficácia da suplementação de própolis bruta nas dietas de frangos de corte sobre as respostas imunes (humoral e celular, peso dos órgãos linfóides e perfil hematológico. Foram utilizados 168 pintos de corte, machos, criados em gaiolas de metabolismo até os 21 dias de idade. As aves foram distribuídas em um delineamento experimental inteiramente casualizado, com seis tratamentos, que consistiram em diferentes níveis de inclusão da própolis (0, 100, 200, 300, 400 e 500 ppm, com sete repetições e quatro aves por unidade experimental. A inclusão de própolis bruta nas rações não afetou (P>0,05 o peso relativo do timo, baço e bolsa cloacal e a produção dos anticorpos sricos contra a doença de Newcastle. Observou-se comportamento quadrático (P0,05. A reação interdigital a fitohemaglutinina apresentou comportamento quadrático em função do tempo e dos níveis de inclusão. De acordo com

  1. Evaluation of the proliferative activity of methanol extracts from six medicinal plants in murine spleen cells

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    Rodrigo Hermes Zandonai

    2010-06-01

    Full Text Available A number of natural compounds have been used as immunomodulatory agents, enabling the function of the immune system to be modified by stimulating or suppressing it. There has been increasing interest in the study of therapeutic action of plant extracts regarding their immunomodulatory activity. The aim of this study was to identify and evaluate the action of extracts of the medicinal plants Calophyllum brasiliense, Ipomoea pes-caprae, Matayba elaeagnoides, Maytenus robusta, Rubus imperialis and Vernonia scorpioides on the development of spleen cells from mice, using the in vitro cellular proliferation assay. The cells, obtained by mechanical rupture of mice spleen (5x10(4 cells/mL, were incubated with methanol extracts (10, 50, 100 and 200 µg/mL and phytohemagglutinin (PHA, 5 µg/mL. The basal control for proliferation consisted of cells alone, while the positive control consisted of cells and PHA. The cell culture was kept at 37 ºC in 5% CO2 for 72 hours, and cell proliferation was revealed by the blue tetrazolium reduction assay (MTT. The results were expressed as percentage of growth and were analyzed using the Kruskal-Wallis and Mann-Whitney tests. The C. brasiliense, I. pes-caprae and M. elaeagnoides extracts showed dose-dependent induction of cell proliferation, with a significant increase in cell proliferation (pVárias substâncias de origem natural têm sido utilizadas como agentes imunomoduladores, permitindo modificar a função do sistema imune e propiciando o estudo de atividades terapêuticas de extratos de plantas. Este trabalho objetivou identificar a atividade imunomodulatória dos extratos de seis plantas medicinais da flora brasileira, Calophyllum brasiliense, Ipomoea pes-caprae, Matayba elaeagnoides, Maytenus robusta, Rubus imperialis e Vernonia scorpioides, sobre a proliferação de células esplênicas de camundongos. As células esplênicas murinas obtidas por ruptura mecânica do baço (5x14³ células/mL foram

  2. Effect of Nutrient Dilution and Glutamine Supplementation on Growth Performance, Small Intestine Morphology and Immune Response of Broilers

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    majid gheshlagh olyayee

    2016-11-01

    vivo cutaneous basophilic hypersensitivity response lectin phytohaemagglutinin (PHA-P and humoral immune response was evaluated by injection of 1 ml of 10 % suspension of sheep red blood cell (SRBC on day 18. Primary immune response was measured after 6 (24 –day-old chick and 12 (30 –day-old chick days of the injection and secondary immune response was assessed on day 36 and 42 experiment. Results and Discussion The results indicated that nutrient dilution and Gln supplementation significantly improved feed conversion ratio (FCR in grower and finisher periods. Gln supplementation increased relative weights of jejunum, small intestine, thymus and bursa of fabricius. The nutrient dilution and Gln significantly affected villi height and crypt depth of jejunum. Gln is an important oxidative fuel for rapidly proliferating cells such as those of the gastrointestinal tract and immune system, reticulocytes, fibroblast. To study humoral immunity, the highest primary and secondary antibody response against Sheep red blood cell (SRBC was seen in diets containing 1.5% Gln and the lowest was seen in control (without Gln supplementation. In cellular immunity determination, 24 h after subcutaneous injection of Phytohemagglutinin-P (PHA-P revealed that Gln supplementation increased toe web thickness. Gln is known to modulate immune function. Glutamine is utilized at a high rate by cells of the immune system in culture and is required to support optimal lymphocyte proliferation and production of cytokines by lymphocytes and macrophages. More recently, Gln has also been shown to have anti-inflammatory effects, modulating cytokine production, both in vitro and in vivo, possibly through decreasing a major transcription factor regulating immune and inflammatory responses. In addition, it has been demonstrated that glutamine can modulate immune response by T cell activation. Therefore the increased toe web thickness after PHA-P injection can be explained by increasing T cell

  3. Isolation and Biological Characteristics of Mesenchymal Stem Cells Derived from Human Placenta Decidua Basalis%人胎盘底蜕膜间充质干细胞的分离及其生物学特性研究

    Institute of Scientific and Technical Information of China (English)

    韩之波; 王有为; 王涛; 池颖; 杨舟鑫; 及月茹; 孟磊; 杨萍; 韩忠朝

    2013-01-01

    Comparing to bone marrow mesenchymal stem cells (MSCs),placenta-derived MSCs have the advantages of adequate sources,low immunogenicity,little risk of viral contamination,and no ethical controversy,and thus possess a better prospect for clinical application.Placental tissue not only includes chorionic and amniotic,but also contains decidua basalis which locate in the maternal placenta surface.The biological characteristics of MSCs isolated from decidua basalis have not been well studied.This study was aimed to investigate the biologic characteristics of placeuta decidua basalis-derived MSC from placenta decidua basalis(DB) by enzymatic digestion.Short tandem repeats (STR) test was used to identify the cells derived from the maternal placenta surface.Growth rate of decidua basalis mesenchymal stem cells (DB-MSC) was measured by MTT.Cell cycle and cell phenotype were detected by flow cytometry.Inducing differentiation was used to evaluate multipotency of DB-MSC.For testing the immunesuppression of DB-MSC,they were co-cultured with peripheral blood mononuclear cells (PBMNC) stimulated by phytohemagglutinin (PHA) and then IFN-γ in the co-cultured media was quantified by ELISA.The results showed that the cells were derived from the maternal placenta by STR analysis.DB-MSC showed typical fibroblast morphology in the culture and were positive for the MSC surface markers:CD90,CD73,CD105,CD44 and negative for CD45,CD11b,and CD34.DB-MSC underwent osteogenic,adipogenic and chondrogenic differentiation in inducing medium.DB-MSC could inhibit the secretion of IFN-γby PBMNC.It is concluded that the cells are isolated from placenta decidua basalis and possess the basic characteristics of MSC.DB-MSC can be an important maternal autologous MSC and may be a safe and effective treatment for immune system diseases,which makes the DB-MSC as an important source of autologous MSC from mother.DB-MSC can be safely for the treatment of the mother's immune system diseases.%胎

  4. Omenn综合征一例临床表型和基因诊断分析%Clinical phenotype and gene diagnostic analysis of Omenn syndrome

    Institute of Scientific and Technical Information of China (English)

    王艳琼; 崔玉霞; 冯杰

    2013-01-01

    Objective Omenn syndrome is a rare autosomal recessive hereditary severe combined immunodeficiency.The purpose of this study was to understand clinical characteristics and genetic mutation type of Omenn syndrome and to improve the recognition of Omenn syndrome among pediatric clinicians.Method One suspected case of severe combined immunodeficiency was found to have pneumonia repeatedly,intractable diarrhea,poor antibiotic treatment effect,lymphadenopathy,hepatosplenomegaly and erythroderma.The patient was diagnosed as having Omenn syndrome by RT-PCR,and the expression of RAG1/RAG2 and gene analysis of RAG1/RAG2 were performed.Result The classification of lymphocyte was CD3 + cells (35.3%),CD19 + cells (0.4%),CD16 + cells (57.6%).After stimulation with phytohemagglutinin (PHA),lymphocyte proliferation of the child was extremely low.Genetic studies showed RAG1 homozygous deletion mutation (2302 del T).He had detectable activated T-lymphocytes with low circulating B-lymphocytes and no evidence of maternal T-cell engrafment as indicated by the short tandem repeat (STR) analysis.Conclusion Omenn syndrome is a severe combined immunodeficiency disease caused by mutations in the RAG1/RAG2 gene.The disease has been reported rarely in China.The clinical manifestations of the disease is early postnatal repeated infections and erythroderma.Mutation analysis of RAG1/RAG2 gene may help to confirm the diagnosis and may be useful in early immune reconstitution and genetic counseling.%目的 Omenn综合征是罕见的常染色体隐性遗传严重联合免疫缺陷病.探讨1例Omenn综合征患儿的临床特征和基因突变类型,以提高临床医师对该病的认识.方法 对1例疑诊严重联合免疫缺陷病患儿进行详细的病史采集和归纳,并采用PCR方法扩增患儿、父母及哥哥RAG1/RAG2基因,PCR产物进行双向序列测定,RT-PCR扩增25个T淋巴细胞受体β链V区(TCRVβ)亚家族并进行克隆谱型分析,短

  5. Effect of intrauterine hepatitis B virus infection on peripheral blood mononuclear cells interferon-γ and interleukin-4 in newborns%乙型肝炎病毒宫内感染对新生儿外周血细胞因子干扰素γ和白介素4的影响

    Institute of Scientific and Technical Information of China (English)

    苏雪松; 彭勋; 陈妍华; 王瑞华; 马洁; 佟艳

    2008-01-01

    Objegtive To observe the effect of intrauterine hepatitis B virus(HBV)infection on peripheral blood mononuclear cells function of secreting interferon-γ and interleukin-4.Methotis Pregnant women were systematically screened for HBsAg and HBeAg when attending the antenatal clinic at the Qinhuangdao Maternal and Child HeMth HospitaL Totally 67 pairs of mothers and infanta were enrolled into this study after obtaining the women's consent.Venous blood samples were collected from the infants within 6 hours after birth and before HBIG injection and HBVac immunization.Blood sample was taken from the mother at or afterthe time when the infant was born.HBV DNA in plasma and PBMC from mothers andtheir newborns were examined using polymerase chmn reaction(PCR).According to HBV DNA in PBMC of newborns.they were divided into two groups.The PBMCs isolated from newborn were cultured with purified HBsAg or phytohemagglutinin(PHA).The supernatant intedeukin-4 and interferon-γ level was measured by using enzyme linked immunosorbent assay(ELISA).Results In 19 newborns PBMC was pesifive for HBV DNA.Maternal PBMCHBVDNApositivitywas associatedwith high rate ofintrauterine HBV infection in the infants(X2=7.58,P<0.01).Compared with the infants whose PBMC HBV DNA was negative,the infanta with PBMC positive for HBV DNA expressed a lower level interferon-γ secretion after purified HBsAg stimulation(t=4.71,P<0.01),however,no significant difference was geen after PHA stimulation (t=1.21,P>0.05).The supematant IL-4 level detected after stimulation with purified HBsAg was higher in the newborns whose PBMC HBV DNA was positive as compared with those negative for PBMC HBV DNA (t=-8.51.P<0.05).The level of IL-4 did not show any significant difference after stimulation with PHA between the PBMC HBV DNA negative and positive groups(t=-2.40,P>0.05).Conclusion Infection with HBV of maternal PBMC is responsible for pefinatM newborn's PBMC HBV infection and it may be an important route of

  6. α-促黑素对银屑病患者外周血单一核细胞产生TNF-α和IL-10的影响%Effects of α-melanocyte-stimulating hormone on the production of TNF-α and IL-10 by peripheral blood mononuclear cells from patients with psoriasis

    Institute of Scientific and Technical Information of China (English)

    毕新岭; 顾军; 张莉; 齐晓虹

    2009-01-01

    目的 探讨α-促黑素(α-MSH)对银屑病患者外周血单一核细胞(PBMC)产生TNF-α、IL-10的影响.方法 标本取自20例寻常性银屑病患者和10例健康献血者,用不同浓度α-MSH处理经过植物血凝素(PHA)刺激的PBMC,用ELISA法检测PBMC培养上清中TNF-α、IL-10水平,应用荧光实时定量PCR分析PBMC中TNF-α mRNA、IL-10 mRNA的表达.结果 银屑病患者PBMC培养上清中TNF-α水平高于正常对照(P<0.01),IL-10水平低于正常对照(P<0.01);各浓度α-MSH均可抑制TNF-α的分泌(P<0.01),浓度为10-13mol/L时作用最强(P<0.01);各浓度α-MSH均可诱导IL-10的分泌(P<0.01,P<0.05),浓度为10-13mol/L时作用最强(P<0.01).浓度为10-13 mol/L的α-MSH下调TNF-α mRNA的表达(P<0.01)、上调IL-10 mRNA的表达(P<0.01).结论 α-MSH可调节银屑病患者TNF-α和IL-10产生.%Objective To investigate the effects of alpha-melanocyte-stimulating hormone (alpha-MSH) on the production of tumor necrosis factor-alpha (TNF-alpha) and intedeukin-10 (IL-10) by peri-pheral blood monohuclear cells (PBMCs) from patients with psoriasis vulgaris. Methods Heparinized peri-pheral blood was obtained from 20 patients with psoriasis vulgaris and 10 healthy human controls. PBMCs were isolated, cultured in complete medium, and stimulated with phytohemagglutinin (PHA) alone, the com-bination of PHA and various concentrations of alpha-MSH, or nothing. After another 48-hour culture, ELISA and real-time PCR were performed to measure the secretion levels of TNF-alpha and IL-10 in the super-natants of cultured PBMCs as well as the mRNA expression levels of TNF-alpha and IL-10 in PBMCs. Results The secretion level of TNF-alpha in the supematants of patient-derived PBMCs stimulated by nothing or PHA alone was significantly higher than that from normal control-derived PBMCs (329.87 ± 99.33 ng/L vs 116.95 ± 37.15 ng/L, 1756.01 ± 183.60 ng/L vs 1287.30 ± 152.36 ng/L, both P<0.01). alpha-MSH of all tested concentrations (10

  7. 间质干细胞对SLE患者外周血T细胞的免疫抑制作用%Immunosuppressive effects of mesenchymal stem cells on peripheral blood T lymphocytes from patients with systemic lupus erythematosus

    Institute of Scientific and Technical Information of China (English)

    殷玉俊; 李晶; 裘影影; 汤郁; 尤海燕; 费晓明; 许文荣

    2009-01-01

    Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs)on peripheral blood T lymphocytes from patients with systemic lupus erythematosus(SLE)in vitro and their potential mechanism.Methods MSCs were isolated from the bone marrow of 3 healthy human volunteers,cultivated and identified.Under phytohemagglutinin(PHA)stimulating,peripheral blood T lymphocytes from 8 patients with SLE were treated with MSCs with the T lymphocyte/MSC ratio being 50:1 in group B and 5:1 in group C or without MSCs(group A).MTT assay was used to detect the proliferation of T lymphocytes.flow cytometry to analyze the expressions of surface markers CD152 and CD28 on T lymphocytes.and real time PCR to measure the mRNA expressions of interleukin-6 and interferon-γ,in the T lymphocytes.Results MSCs could markedly inhibit the proliferation of T lymphocytcs.The proliferation of T lymphocytes expressed as absorbance value at 570 nm was 0.484±0.032 in group B.0.308±0.025 in group C,significantly lower than that in group A(0.765±0.036,both P<0.05),and significant difference was also observed between group C and B(P<0.05).In the case of the percentage of CD28 positive T lymphocytes.group B and C were significantly lower than group A(60.39%±3.92%and 45.05%±3.46%vs 74.73%±3.74%,both P<0.05),and group B significantly differed from group C(P<0.05).MSCs had no obvious effect on the expression of CD152 on T lymphocytes,but significantly suppressed the mRNA expression of interleukin-6 and interferon-γ(both P<0.05).and the suppressive effect was enhanced with the incrgase in MSC count.ConclusionsMSCs exert an immunosuppressive effect on T lymphocytes from patients with SLE,likely through inhibiting the proliferation,CD28 expression,interleukin-6 and interferon-γ mRNA expression of T lymphocytes.%目的 探讨体外骨髓间质干细胞(MSC)对SLE患者外周血T淋巴细胞(T细胞)的免疫调节作用及可能机制.方法 分离培养健康人骨髓MSC并鉴

  8. Effect of Dentritic Cells from Malignant Pleural Effusions on Tumor Infiltration Lymphocytes%恶性胸腔积液来源树突状细胞对自体肿瘤浸润性淋巴细胞的作用

    Institute of Scientific and Technical Information of China (English)

    曾波航; 陈静琦; 黄慧

    2011-01-01

    Objective Dentritic cells (DCs) from malignant pleural effusions of patients with lung cancer were isolated and induced.Effect of DCs on proliferation and cytotoxicity of tumor infiltration lymphocytes (TILs) from origin malignant pleural effusions was studied.Methods Pleural effusion mononuclear cells were obtained from 16 patients with lung cancer.Cells were separated by density gradient and magnetic cell sorting system.DCs were induced by interleukin-4 (IL-4), tumor necrosis factor-α(TNF-α), and granulocyte-macrophage colony stimulating.DCs were observed by optical microscope, electronic microscope and flow cytometry respectively.TILs from origin malignant pleural effusions were induced by IL-2 and phytohemagglutinin.The cancer cells were separated with HEAl25 magnetic cell sorting system.The ability of stimulating the proliferation of TILs was detected with 3H-thymidine.The anti-tumor effect of TILs was measured with MTT method.Results The mature DCs from malignant pleural effusions of patients with lung cancer could be induced.The typical morphology in DCs was observed with optical and electronic microscopes.DCs expressed high-level surface phenotypes, including HLA-ABC, HLA-DR, CD86, CD54 and CD83, CDla.DCs can increase about 1.7 times proliferation of TILs.The cytotoxicity of TILs stimulated with DCs was increased from (31.80 ± 14.05) % to (51.89 ± 13.27) %.Conclusion The mature DCs can be induced from malignant pleural effusions from patients with lung cancer.The DCs can stimulate TILs from the origin malignant pleural effusion and increase its proliferation ability and cytotoxicity to cancer cells.%目的 探讨恶性胸腔积液来源 DCs(dentritic cells,DCs)对自体肿瘤浸润性淋巴细胞(tumorinfiltration lymphocytes,TILs)增殖及杀伤肿瘤细胞能力的影响.方法 用体外培养方法从16例肺癌患者恶性胸腔积液来源分离单个核细胞,再用密度梯度离心辅以免疫磁珠分选细胞,用白细胞介素4(IL-4)、

  9. Inhibitory Effect of Human Umbilical Cord-derived Mesenchymal Stem Cells on Interleukin-17 Production in Peripheral Blood T Cells from Spondyloarthritis Patients%人脐带间充质干细胞对脊柱关节炎患者外周血T细胞产生IL-17的抑制作用

    Institute of Scientific and Technical Information of China (English)

    黄志芳; 朱剑; 吕双红; 张江林; 陈显达; 杜丽欣; 杨志岗; 宋亚昆; 吴东颖

    2013-01-01

    .82)% vs (0.75 ±0.25)% , P<0.01; CD3 + γ8TCR + IL-17 + cells (0.30 ±0. 10) % vs (0.06 ±0.02) % , P<0.01]. After co-culture of PBMNC in patients with hUCMSC, the increased proportions of CD3+ CD4 + IL-17+ cells and CD3 + 78TCR+ IL-17+ cells in SpA patients were inhibited significantly by hUCMSC[CD3+CD4+ IL-17+ cells (3.42±0.82)% vs (1.81 ±0.59)% (P<0.01); CD3+ 78TCR+ IL-17+ cells (0.30±0. 10)% vs (0.16 ±0.06)% (P<0.01]. In response to phytohemagglutinin (PHA, μg/ml), PBMNC from SpA patients secreted more IL-17 than that from healthy control [ (573. 95 ± 171. 68) pg/ml vs (115.53 ±40.41) pg/ml (P<0.01)]. In the presence of hUCMSC, PBMNC of SpA patients produced less amount of IL-17 [ (573.95 ± 171.68)pg/ml vs (443.20 ±147.94) pg/ml, (P<0.01)]. It is concluded that the IL-17 production in peripheral blood T cells from SpA patients can be inhibited by hUCMSC, which have therapeutic potential for SpA.

  10. Inhibitory effects of human umbilical cord-derived mesenchymal stem cells on proliferation of peripheral blood mononuclear cells from spondyloarthritis patients%人脐带间充质干细胞对脊柱关节炎患者外周血单个核细胞体外增殖的抑制作用

    Institute of Scientific and Technical Information of China (English)

    黄志芳; 吕双红; 朱剑; 杨志岗; 宋亚昆; 杜丽欣; 陈显达; 胡海旭; 吴东颖

    2013-01-01

    目的 探讨人脐带间充质干细胞(hUCMSC)对脊柱关节炎(SpA)患者外周血单个核细胞(PBMC)体外增殖的抑制作用.方法 采用随机区组设计或配对设计,将12例SpA患者的PBMC与hUCMSC共培养或单独培养,CCK-8法检测PBMC增殖,并以流式细胞术检测其细胞周期分布;同时将hUCMSC的作用与SpA患者临床资料进行相关分析.结果 hUCMSC抑制SpA患者PBMC体外增殖,比例越大抑制作用越强(P<0.01),直接接触共培养的抑制作用强于Transwell小室培养(57%±17%比32%±12%),两组比较差异有统计学意义(P<0.01);hUCMSC使处于G1期的PBMC增多(86%±3%比68%±5%),处于(S+G2)期的PBMC减少(8%±3%比26%±5%),两组比较差异有统计学意义(P<0.01);hUCMSC的抑制作用与SpA患者的临床资料无相关性.结论 hUCMSC能够抑制SpA患者PBMC的体外增殖,在SpA的临床治疗中具有潜在的应用前景.%Objective To explore the inhibitory effects of human umbilical cord-derived mesenchymal stem cells (hUCMSC) on the proliferation of peripheral blood mononuclear cells (PBMC) from spondyloarthritis (SpA) patients.Methods A total of 12 SpA patients at Chinese PLA General Hospital were recruited from May 2012 to October 2012.Information on demographic characteristics,disease and functional activity was collected.Isolated PBMC were stimulated by phytohemagglutinin (PHA,1 μg/ml) in the presence or absence of hUCMSC.The proliferation of hUCMSC was suppressed by irradiation with Co60(30 Gy) before co-culturing with PBMC.The proliferation of PBMC was determined by Cell Counting Kit-8(CCK-8).Cell cycle profiles of PBMC were analyzed by flow cytometry.The association of inhibitory effect of hUCMSC with the disease and functional activity of SpA patients was examined.Results After coculturing with hUCMSC by cell-to-cell contact for 5 days,the proliferation of PBMC stimulated by PHA (1 μg/ml)was significantly inhibited by hUCMSC in a dose

  11. 酵解乳化饲料对哺乳母猪生产性能、免疫力及饲粮养分消化的影响%Effect of Glycolysis and Emulsification Feed on Performance, Immunity and Dietary Nutrient Digestion of Lactating Sows

    Institute of Scientific and Technical Information of China (English)

    朱元召; 孟秀丽; 程金龙; 尹龙

    2015-01-01

    本试验旨在研究酵解乳化饲料对哺乳母猪生产性能、免疫力及饲粮养分消化的影响。选取胎次、预产期相近的长×大母猪160头,随机分成5个组(对照组、麦芽糊精和葡萄糖组、发酵豆粕组、乳化脂肪组、酵解乳化饲料组),每组4个重复,每重复8头。对照组母猪饲喂基础饲粮;麦芽糊精和葡萄糖组饲粮中添加1.5%麦芽糊精+1.5%葡萄糖;发酵豆粕组、乳化脂肪组、酵解乳化饲料组饲粮中分别添加5.0%发酵豆粕、3.0%乳化脂肪、10.0%酵解乳化饲料。试验从妊娠108 d开始,妊娠114 d分娩,至仔猪28日龄断奶时结束,共34 d。结果表明:1)与对照组、麦芽糊精和葡萄糖组、发酵豆粕组、乳化脂肪组相比,酵解乳化饲料组母猪的平均日采食量分别提高了7.08%(P0.05)、5.78%(P>0.05)、4.83%(P>0.05),母猪断奶后发情间隔分别缩短了26.58%(P0.05)、8.20%( P0.05),28日龄仔猪成活率分别提高了6.14%(P0.05)、2.09%(P>0.05)、2.83%( P>0.05)。2)与对照组相比,酵解乳化饲料组母猪血清免疫球蛋白( Ig) A、IgG、IgM含量,植物血凝素淋巴细胞转化率分别提高了23.93%( P0.05), 5.78% (P>0.05) and 4.83% (P>0.05), after weaning to estrus interval of lactating sows was short by 26.58% (P0.05) , 8.20% ( P0.05), and survival rate of 28-day-old piglets was increased by 6.14% (P0.05), 2.09% (P>0.05) and 2.83% (P>0.05), respectively. 2) The content of serum immunoglobulin ( Ig) A, IgG, IgM and phytohemagglutinin ( PHA) lymphocyte transformation rate of lactating sows in glycol-ysis and emulsification feed group were increased by 23.93% (P<0.01), 39.17% (P<0.05), 19.28% (P<0.05) and 17.64% ( P<0.01) , respectively, while the content of serum cortisol was decreased by 5.97% ( P<0.05) than those in the control group. 3) Compared with the control group, the apparent digestibility of ether extract, organic matter and gross energy in glycolysis and emulsification feed group was

  12. T cell proliferative response and antibody formation to citrullinated collagen type Ⅱ in patients with rheumatoid arthritis%类风湿关节炎患者瓜氨酸化Ⅱ型胶原的T细胞增殖反应和抗体生成

    Institute of Scientific and Technical Information of China (English)

    田昕; 赵义; 栗占国

    2009-01-01

    fetal bovine serum RPMI 1640 fluid, phytohemagglutinin (PHA), or citrullinated buffer as controls. Cellular reactivity levels against Cit-C Ⅱ and C Ⅱ were investigated by measuring the proliferation of PBMCs to calculate the stimulation index (SI). ELISA was used to detect the presence of antibodies against Cit-C Ⅱ and C Ⅱ. Results The positive rate of T cell proliferative response to Cit-C Ⅱ of the RA group was 32.4% (11/34), significantly higher than that of the control group (0,P<0.05). The positive rate of T cell proliferative response to C Ⅱ of the RA group was 35.3% (12/34), significantly higher than that of the control group (11.1%, P<0.05). Interestingly, Cit-C Ⅱ could not elicit a better T cell proliferative response than C Ⅱ did. In the RA patients, the anti-Cit-C Ⅱ antibody positive rate was 52.9% (18/34), significantly higher than that of the anti-C Ⅱ antibody positive rate [32.4% (11/34), P<0.05). The serum IgG anti-C Ⅱ antibody positive rate of the patients with positive T cell responses to C Ⅱ was 58.3% (7/12), significantly higher than that of the patients with negative T cell responses to C Ⅱ [18.2% (4/22), P <0.05]. The serum lgG anti-C Ⅱ antibody positive rate of the patients with positive T cell responses to Cit-C Ⅱ was 72.7% (8/11), significantly higher than that of the patients with negative T cell responses to Cit-C Ⅱ [43.5% (10/23), P<0.05]. Conclusion The recognition of Cit-C Ⅱ by circulating IgG antibodies is a RA-specific serological phenomenon. The formation of C Ⅱ antibody in the RA patients may be related to B cell activation mediated by C Ⅱ specific T cells. The role of Cit-C Ⅱ in RA cellular immune need be further studied.

  13. Shock waves co-stimulate T-cell proliferation and interleukin-2 expression through ATP release, P2 receptor and p38 mitogen activated protein kinase activation%冲击波通过ATP释放、P2受体及激活p38MAPK激酶促进T细胞增殖和分泌白细胞介素2

    Institute of Scientific and Technical Information of China (English)

    于铁成; 赵毅; 陈玮伦; 金安; 刘建国

    2007-01-01

    Source Inc., Camarillo, CA); ATP Bioluminescence Assay Kit CLS Ⅱ (Roche Diagnostics GmbH,Mannheim, Germany).METHODS: The experiment was carried out in the Orthopedic Laboratory of the First Clinical Hospital in Jilin University from January 2005 to December 2006. ①An Extracorporeal Shockwave Lithotripter (at 7 kV generator voltage, 0.3 μF capacitance, 23 MPa positive pressure, 0.18 mJ/mm2 energy flux density) was applied for LDSWs treatment ranging from 50 to 400 impulses. ②ATP release into the culture supernatant from Jurkat T-cells or human peripheral blood mononuclear cells (PBMCs) was determined with a specific ATP Bioluminescence Assay Kit. ③Negative control group excluded antagonist or inhibitor. Human PBMCs were used to determine the effect of LDSWs on activated T-lymphocyte proliferation. Human Jurkat cells were used to study the effects of LDSWs on IL-2 expression. Expression and phosphorylation of p38 MAPK in Jurkat T-cell were measured by Western Immunoblotting with anti-p38 MAPK antibodies and anti-p38 MAPK phospho-specific antibodies that recognized the phosphorylation (on Thr180/Tyr182).MAIN OUTCOME MEASURES: extra-cellular ATP release, IL-2 expression in cell suspension, cellular proliferation and the phosphorylation of p38 MAPK.RESULTS: ①ATP release under the condition without LDSWs was obviously lower than that with LDSWs of 100, 150,200, 250, 300, 360 and 400 impulses (P < 0.01), and ATP release increased with the LDSWs impulse.②Compared with negative control group, the additions of apyrase, KN-62 or suramin attenuated the 3H-TdR incorporation of the phytohemagglutinin-stimulated PBMCs or CD3/CD28-stimulated Jurkat T-cells, which were effected with LDSWs of 100,150, 200, 250, 300, 330 impulses at 0.18 mJ/mm2 (P< 0.01). IL-2 expression in the cellular supernatant was also significant increased (P < 0.01). ATPase, KN-62 or suramin all decreased the effect of LDSWs on p38 MAPK of Jurkat T-cells.CONCLUSION: ①LDSWs deform cellular membranes