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Sample records for basophils

  1. Basophils and skin disorders.

    Science.gov (United States)

    Borriello, Francesco; Granata, Francescopaolo; Marone, Gianni

    2014-05-01

    Since their discovery in 1879, basophils have been viewed as circulating blood granulocytes with limited immune function. New research tools for their functional analysis in vivo have revealed previously unrecognized roles for basophils in several skin disorders. Human basophils infiltrate different skin lesions and have been implicated in the pathogenesis of diseases ranging from chronic idiopathic urticaria to systemic lupus erythematosus. In mouse models, basophils participate in IgE-mediated chronic allergic inflammation of the skin and have a protective role in tick infestation. In this review, we discuss critical advances in our understanding of basophil biology and their roles in the pathophysiology of skin disorders.

  2. Basophil stimulation and signaling pathways.

    Science.gov (United States)

    Knol, Edward F; Gibbs, Bernhard F

    2014-01-01

    Despite growing use of flow cytometry to analyze the functional characteristics of primary basophils the intracellular signaling cascades that control their ability to elaborate various inflammatory mediators and cytokines remain comparatively obscure. Additionally, some studies require the analysis of pro-allergic and inflammatory mediators, such as histamine, LTC4, and various basophil-derived cytokines (e.g., IL-4 and IL-13). Elucidation of intracellular signaling proteins by Western blotting, cytosolic free calcium concentration by spectrofluorophotometry, and detection of mediator releases, as well as analysis of gene expressions by RT-PCR, generally require relatively large numbers of purified basophils. In selected assays, flow cytometry can enable the analysis of relatively low cell numbers and purity for the expression of intracellular signaling proteins or measurement of cytosolic free calcium concentrations by basophil-specific gating strategies. Unfortunately, many aspects of signal transduction relevant to human basophils cannot be readily extrapolated from the use of basophil or mast cell lines. This chapter therefore focuses on how to employ primary human basophils for studying mediator releases and signaling characteristics.

  3. Genetic analysis of basophil function in vivo

    OpenAIRE

    Sullivan, Brandon M.; Liang, Hong-Erh; Bando, Jennifer K.; Wu, Davina; Cheng, Laurence E.; McKerrow, James K.; Allen, Christopher D.C.; Locksley, Richard M.

    2011-01-01

    Contributions by basophils to allergic and helminth immunity remain incompletely defined. Using sensitive IL-4 reporter alleles, we demonstrate that basophil IL-4 production occurs by a CD4+ T cell-dependent process restricted to affected peripheral tissues. We genetically marked and specifically deleted basophils and demonstrate that basophils do not mediate TH2 priming in vivo. Two-photon imaging confirmed that basophils do not interact with antigen-specific T cells in lymph nodes, but can ...

  4. Genetic analysis of basophil function in vivo.

    Science.gov (United States)

    Sullivan, Brandon M; Liang, Hong-Erh; Bando, Jennifer K; Wu, Davina; Cheng, Laurence E; McKerrow, James K; Allen, Christopher D C; Locksley, Richard M

    2011-06-01

    Contributions by basophils to allergic and helminth immunity remain incompletely defined. Using sensitive interleukin 4 (Il4) reporter alleles, we demonstrate here that basophil IL-4 production occurs by a CD4(+) T cell-dependent process restricted to the peripheral tissues affected. We genetically marked and achieved specific deletion of basophils and found that basophils did not mediate T helper type 2 (T(H)2) priming in vivo. Two-photon imaging confirmed that basophils did not interact with antigen-specific T cells in lymph nodes but engaged in prolonged serial interactions with T cells in lung tissues. Although targeted deletion of IL-4 and IL-13 in either CD4(+) T cells or basophils had a minimal effect on worm clearance, deletion from both lineages demonstrated a nonredundant role for basophil cytokines in primary helminth immunity.

  5. Basophil-activation tests in hymenoptera allergy

    NARCIS (Netherlands)

    Dubois, Anthony E. J.; van der Heide, Sicco

    2007-01-01

    The measurement of basophil-activation markers may be useful in detecting IgIE-mediated sensitization but the relevance for application of the basophil-activation test in prediction of clinical reactivity in Hymenoptera allergy is very limited. For this reason, this test currently has no established

  6. Human cord blood derived immature basophils show dual characteristics, expressing both basophil and eosinophil associated proteins

    OpenAIRE

    Jeanette Grundström; Jenny M Reimer; Sofia E Magnusson; Gunnar Nilsson; Sara Wernersson; Lars Hellman

    2012-01-01

    Basophils are blood cells of low abundance associated with allergy, inflammation and parasite infections. To study the transcriptome of mature circulating basophils cells were purified from buffy coats by density gradient centrifugations and two-step magnetic cell sorting. However, after extensive analysis the cells were found to be transcriptionally inactive and almost completely lack functional mRNA. In order to obtain transcriptionally active immature basophils for analysis of their transc...

  7. Human basophils express interleukin-4 receptors

    Energy Technology Data Exchange (ETDEWEB)

    Valent, P.; Besemer, J.; Kishi, K.; Di Padova, F.; Geissler, K.; Lechner, K.; Bettelheim, P. (Univ. of Vienna (Austria))

    1990-11-01

    Interleukin-4 (IL-4), a multipotential lymphokine reputed to play an important role in the regulation of immune responses, interacts with a variety of hemopoietic target cells through specific cell surface membrane receptors. The present study was designed to investigate whether human basophils express IL-4 binding sites. For this purpose, basophils were enriched to homogeneity (93% and 98% purity, respectively) from the peripheral blood of two chronic granulocytic leukemia (CGL) donors using a cocktail of monoclonal antibodies (MoAbs) and complement. Purified basophils bound 125I-radiolabeled recombinant human (rh) IL-4 in a specific manner. Quantitative binding studies and Scatchard plot analysis revealed the presence of a single class of high affinity IL-4 binding sites (280 +/- 40 sites per cell in donor 1 and 640 +/- 45 sites per cell in donor 2) with an apparent dissociation constant, kd, of 7.12 x 10(-11) +/- 2.29 x 10(-11) and 9.55 +/- 3.5 x 10(-11) mol/L, respectively. KU812-F, a human basophil precursor cell line, was found to express a single class of 810 to 1,500 high affinity IL-4 binding sites with a kd of 2.63 to 5.54 x 10(-10) mol/L. No change in the numbers or binding constants of IL-4 receptors was found after exposure of KU812-F cells to rhIL-3 (a potent activator of basophils) for 60 minutes. No effect of rhIL-4 on 3H-thymidine uptake, release or synthesis of histamine, or expression of basophil differentiation antigens (Bsp-1, CD11b, CD25, CD40, CD54) on primary human CGL basophils or KU812-F cells was observed.

  8. Newly appreciated roles for basophils in allergy and protective immunity.

    Science.gov (United States)

    Karasuyama, H; Obata, K; Wada, T; Tsujimura, Y; Mukai, K

    2011-09-01

    Basophils are evolutionarily conserved in many animal species, in spite of the fact that they account for basophils have an indispensable and nonredundant role in vivo, even though they show some phenotypic similarity with tissue-resident mast cells. However, their functional significance remained uncertain long after Paul Ehrlich discovered them as blood-circulating cells with basophilic granules more than 130 years ago. The study of basophils has been far behind that of mast cells, owing to the rarity of basophils and the paucity of tools for their detection and functional analysis. Recent development of novel analytical tools, including basophil-depleting antibodies and genetically engineered mice deficient only in basophils, has greatly advanced basophil research and illuminated previously unrecognized roles of basophils. We now appreciate that basophils and mast cells play distinct roles in immune responses. Basophils have crucial roles in the development of acute and chronic allergic responses, the protective immunity against ecto- and endoparasites, and the regulation of acquired immunity, including the augmentation of humoral memory responses and the initiation of Th2 responses. Thus, basophils are no longer the neglected minority and are key players in the immune system.

  9. Enhancement of basophil apoptosis by olopatadine and theophylline.

    Science.gov (United States)

    Kawakami, Ayako; Suzukawa, Maho; Koketsu, Rikiya; Komiya, Akiko; Ohta, Ken; Yamamoto, Kazuhiko; Yamaguchi, Masao

    2008-01-01

    Regulation of basophil survival is an important aspect in the pathogenesis of allergic inflammation associated with local accumulation of basophils. However, pharmacologic modulation of basophil survival is largely unknown except for the apoptosis-enhancing effect of glucocorticoids. We tested the effects of two anti-allergic and anti-asthmatic drugs, olopatadine and theophylline, on basophil survival. Basophils were highly purified from normal human peripheral blood. Apoptosis was analyzed by flow cytometry using annexin V staining or another staining method that detected alterations in the mitochondrial transmembrane potential. In addition to the conventional method using annexin V, basophil apoptosis was successfully established by analysis of the mitochondrial transmembrane potential. Olopatadine decreased the number of live basophils, and they induced apoptosis of basophils during culture. The decline in live basophils was induced by olopatadine even when low doses of IL-3 were included in the culture medium. Theophylline also affected basophil apoptosis and induced a decrease in the number of live basophils. Basophil apoptosis was enhanced by both olopatadine and theophylline. This effect may partly explain the pharmacologic basis of why these drugs are effective on allergic diseases.

  10. Measuring histamine and cytokine release from basophils and mast cells

    DEFF Research Database (Denmark)

    Jensen, Bettina M; Falkencrone, Sidsel; Skov, Per S

    2014-01-01

    Basophils and mast cells are known for their capability to release both preformed and newly synthesized inflammatory mediators. In this chapter we describe how to stimulate and detect histamine released from basophils in whole blood, purified basophils, in vitro cultured mast cells, and in situ...

  11. Spontaneous and cytokine induced basophil adhesion evaluated by microtiter assay

    DEFF Research Database (Denmark)

    Quan, Sha; Poulsen, Lars K; Reimert, Claus Michael;

    2002-01-01

    We have developed a microtiter assay for evaluating basophil spontaneous adhesion to extracellular matrix (ECM) proteins exemplified by fibronectin and cytokine induced basophil adhesion to bovine serum albumin (BSA). The percentage of basophils adhering to either ECM or BSA was quantified...

  12. Diagnostic tests based on human basophils

    DEFF Research Database (Denmark)

    Kleine-Tebbe, Jörg; Erdmann, Stephan; Knol, Edward F;

    2006-01-01

    -maximal responses, termed 'intrinsic sensitivity'. These variables give rise to shifts in the dose-response curves which, in a diagnostic setting where only a single antigen concentration is employed, may produce false-negative data. Thus, in order to meaningfully utilize the current basophil activation tests...

  13. Human cord blood derived immature basophils show dual characteristics, expressing both basophil and eosinophil associated proteins.

    Directory of Open Access Journals (Sweden)

    Jeanette Grundström

    Full Text Available Basophils are blood cells of low abundance associated with allergy, inflammation and parasite infections. To study the transcriptome of mature circulating basophils cells were purified from buffy coats by density gradient centrifugations and two-step magnetic cell sorting. However, after extensive analysis the cells were found to be transcriptionally inactive and almost completely lack functional mRNA. In order to obtain transcriptionally active immature basophils for analysis of their transcriptome, umbilical cord blood cells were therefore cultured in the presence of interleukin (IL-3 for 9 days and basophils were enriched by removing non-basophils using magnetic cell sorting. The majority of purified cells demonstrated typical metachromatic staining with Alcian blue dye (95% and expression of surface markers FcεRI and CD203c, indicating a pure population of cells with basophil-like phenotype. mRNA was extracted from these cells and used to construct a cDNA library with approximately 600 000 independent clones. This library served as tool to determine the mRNA frequencies for a number of hematopoietic marker proteins. It was shown that these cells express basophil/mast cell-specific transcripts, i.e. β-tryptase, serglycin and FcεRI α-chain, to a relatively low degree. In contrast, the library contained a high number of several eosinophil-associated transcripts such as: major basic protein (MBP, charcot leyden crystal (CLC, eosinophil cationic protein (ECP, eosinophil derived neurotoxin (EDN and eosinophil peroxidase (EPO. Out of these transcripts, MBP and EPO were the most frequently observed, representing 8% and 3.2% of the total mRNA pool, respectively. Moreover, in a proteome analysis of cultured basophils we identified MBP and EPO as the two most prominent protein bands, suggesting a good correlation between protein and mRNA analyses of these cells. The mixed phenotype observed for these cells strengthens the conclusion that

  14. Human cord blood derived immature basophils show dual characteristics, expressing both basophil and eosinophil associated proteins.

    Science.gov (United States)

    Grundström, Jeanette; Reimer, Jenny M; Magnusson, Sofia E; Nilsson, Gunnar; Wernersson, Sara; Hellman, Lars

    2012-01-01

    Basophils are blood cells of low abundance associated with allergy, inflammation and parasite infections. To study the transcriptome of mature circulating basophils cells were purified from buffy coats by density gradient centrifugations and two-step magnetic cell sorting. However, after extensive analysis the cells were found to be transcriptionally inactive and almost completely lack functional mRNA. In order to obtain transcriptionally active immature basophils for analysis of their transcriptome, umbilical cord blood cells were therefore cultured in the presence of interleukin (IL)-3 for 9 days and basophils were enriched by removing non-basophils using magnetic cell sorting. The majority of purified cells demonstrated typical metachromatic staining with Alcian blue dye (95%) and expression of surface markers FcεRI and CD203c, indicating a pure population of cells with basophil-like phenotype. mRNA was extracted from these cells and used to construct a cDNA library with approximately 600 000 independent clones. This library served as tool to determine the mRNA frequencies for a number of hematopoietic marker proteins. It was shown that these cells express basophil/mast cell-specific transcripts, i.e. β-tryptase, serglycin and FcεRI α-chain, to a relatively low degree. In contrast, the library contained a high number of several eosinophil-associated transcripts such as: major basic protein (MBP), charcot leyden crystal (CLC), eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO). Out of these transcripts, MBP and EPO were the most frequently observed, representing 8% and 3.2% of the total mRNA pool, respectively. Moreover, in a proteome analysis of cultured basophils we identified MBP and EPO as the two most prominent protein bands, suggesting a good correlation between protein and mRNA analyses of these cells. The mixed phenotype observed for these cells strengthens the conclusion that eosinophils and

  15. Basophil competence during hookworm (Necator americanus) infection.

    Science.gov (United States)

    Pritchard, David I; Hooi, Doreen S W; Brown, Alan; Bockarie, Moses J; Caddick, Rebecca; Quinnell, Rupert J

    2007-11-01

    A popular hypothesis to explain parasite survival in the presence of a pronounced T helper 2 phenotype in helminth-parasitized populations has been Fc epsilonRI blockade by parasite-induced polyclonal IgE. To begin to test the hypothesis that Fc epsilonRI-bearing cells would be refractory to activation in parasitized populations, we investigated basophil function in 43 individuals from a hookworm endemic area. Study individuals had high levels of total IgE and eosinophilia and a mean hookworm burden of 2,257 epg. Basophils from all members of this parasitized population were shown to release histamine to a number of agonists, including anti IgE and a hookworm allergen, calreticulin. These data would indicate that Fc epsilonRI blockade at the level of the basophil did not occur in this parasitized population despite the presence of possible immunologic blocking agents. This would suggest that this effector arm of the T helper 2 phenotype remains operative in infected populations.

  16. Adherence of human basophils to cultured umbilical vein endothelial cells.

    OpenAIRE

    1988-01-01

    The mechanism by which circulating human basophils adhere to vascular endothelium and migrate to sites of allergic reactions is unknown. Agents have been identified which stimulate the adherence of purified basophils to cultured human umbilical vein vascular endothelial cells (HuVEC). Treatment of HuVEC with interleukin 1, tumor necrosis factor (TNF), bacterial endotoxin, and 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in time and dose-dependent increases of adhesiveness for basophils...

  17. GATA-1 regulates the generation and function of basophils.

    Science.gov (United States)

    Nei, Yuichiro; Obata-Ninomiya, Kazushige; Tsutsui, Hidemitsu; Ishiwata, Kenji; Miyasaka, Masayuki; Matsumoto, Kenji; Nakae, Susumu; Kanuka, Hirotaka; Inase, Naohiko; Karasuyama, Hajime

    2013-11-12

    Developmental processes of hematopoietic cells are orchestrated by transcriptional networks. GATA-1, the founding member of the GATA family of transcription factors, has been demonstrated to play crucial roles in the differentiation of erythroid cells, magakaryocytes, eosinophils, and mast cells. However, the role of GATA-1 in basophils remains elusive. Here we show that basophils abundantly express Gata1 mRNAs, and that siRNA-mediated knockdown of Gata1 resulted in impaired production of IL-4 by basophils in response to the stimulation with IgE plus antigens. ΔdblGATA mice that carry the mutated Gata1 promoter and are widely used for functional analysis of eosinophils owing to their selective loss of eosinophils showed a decreased number of basophils with reduced expression of Gata1 mRNAs. The number of basophil progenitors in bone marrow was reduced in these mice, and the generation of basophils from their bone marrow cells in culture with IL-3 or thymic stromal lymphopoietin was impaired. ΔdblGATA basophils responded poorly ex vivo to stimulation with IgE plus antigens compared with wild-type basophils as assessed by degranulation and production of IL-4 and IL-6. Moreover, ΔdblGATA mice showed impaired responses in basophil-mediated protective immunity against intestinal helminth infection. Thus, ΔdblGATA mice showed numerical and functional aberrancy in basophils in addition to the known deficiency of eosinophils. Our findings demonstrate that GATA-1 plays a key role in the generation and function of basophils and underscore the need for careful distinction of the cell lineage responsible for each phenotype observed in ΔdblGATA mice.

  18. Pros and Cons of Clinical Basophil Testing (BAT)

    NARCIS (Netherlands)

    Hoffmann, Hans Jürgen; Knol, Edward F; Ferrer, Martha; Mayorga, Lina; Sabato, Vito; Santos, Alexandra F; Eberlein, Bernadette; Nopp, Anna; MacGlashan, Donald

    2016-01-01

    PURPOSE OF REVIEW: We review basophil testing by flow cytometry with an emphasis on advantages and disadvantages. RECENT FINDINGS: There are many tools available to assess the presence and severity of allergic diseases in patients. For 50 years, peripheral blood basophils have been used as tools to

  19. Basophil leucocyte response in leprosy with lepromin-A

    Directory of Open Access Journals (Sweden)

    Okhandiar R

    1993-01-01

    Full Text Available Study of ′Basophil Leucocyte Response′ in two polar groups of leprosy patients (tuberculoid and lepromatous was undertaken employing Lepromin-A (Armadillo by the method of Open Window Technique of Rebuck. Significant infiltration of Basophils at the test side is demonstrated both at 48 hours and at 72 hours in lepromatous leprosy as compared to the control groups i.e., in placebo and tuberculin positive healthy volunteers as well as against tuberculoid leprosy group (p value being less than 0.05 and 0.001. The basophils also go into degeneration and degranulation at 72 hours. . The role of basophils in the different types of allergic reactions is discussed in context of above finding. The possible effect of high infiltration of basophils in lepromatous leprosy with its subsequent release of histamine and other factors by degranulation, and its possible role in immuno-regulation through specialised suppressor cells is postulated in the light of available data.

  20. Klinefelter syndrome and acute basophilic leukaemia--case report.

    Science.gov (United States)

    Ljubić, Nives; Lang, Nada; Skelin, Ika Kardum; Lasan, Ruzica; Dominis, Mara; Perković, Leila; Zupanić-Krmek, Dubraka; Grgurević-Batinica, Anita

    2010-06-01

    Patients with 47, XXY karyotype (Klinefelter syndrome) appear to have increased risk of developing cancer, especially male breast cancer, germ cell tumours and non Hodgkin lymphomas, but rarely acute myeloid leukaemia. We report a patient with acute basophilic leukaemia with 47, XXY karyotype in both the tumour and constitutional cells. Acute basophilic leukaemia is very rare disease comprising less than 1% of all acute myeloid leukaemias. Morphological characteristic of leukaemic blast cells is moderately basophilic cytoplasm containing a variable number of coarse basophilic granules. The most characteristic cytochemical reaction is metachromatic positivity with toluidine blue. Blast are myeloperoxidase negative. Also leukemic blasts express myeloid and monocyte markers. There is no consistent chromosomal abnormality identified in this leukaemia. This is the first reported case of acute basophilic leukaemia in patient with Klinefelter syndrome. In this article the medical history of the patient is given and the possible connection between Klinefelter syndrome and acute myeloid leukaemia is discussed.

  1. Mast cells and basophils in cutaneous immune responses.

    Science.gov (United States)

    Otsuka, A; Kabashima, K

    2015-02-01

    Mast cells and basophils share some functions in common and are generally associated with T helper 2 (Th2) immune responses, but taking basophils as surrogate cells for mast cell research or vice versa for several decades is problematic. Thus far, their in vitro functions have been well studied, but their in vivo functions remained poorly understood. New research tools for their functional analysis in vivo have revealed previously unrecognized roles for mast cells and basophils in several skin disorders. Newly developed mast cell-deficient mice provided evidence that mast cells initiate contact hypersensitivity via activating dendritic cells. In addition, studies using basophil-deficient mice have revealed that basophils were responsible for cutaneous Th2 skewing to haptens and peptide antigens but not to protein antigens. Moreover, human basophils infiltrate different skin lesions and have been implicated in the pathogenesis of skin diseases ranging from atopic dermatitis to autoimmune diseases. In this review, we will discuss the recent advances related to mast cells and basophils in human and murine cutaneous immune responses.

  2. Basophilic deposits on the cupula: preliminary findings describing the problems involved in studies regarding the incidence of basophilic deposits on the cupula.

    Science.gov (United States)

    Naganuma, H; Kohut, R I; Ryu, J H; Tokumasu, K; Okamoto, M; Fujino, A; Hoshino, I; Arai, M

    1996-01-01

    In this study, the possibility of whether basophilic deposits adhered to the cupulas in the semicircular canals was investigated histologically. Results indicated that basophilic deposits were present in all three cupulas of the semicircular canals. The overall incidence of basophilic deposits in the superior, lateral and posterior semicircular canal cupulas was 26%, 41% and 37%, respectively. The incidence of basophilic deposits bound to the cupulas increased with age. The possible origin of these basophilic deposits on the cupulas and the increased incidence of basophilic deposits with increasing age are discussed.

  3. Signaling transduction pathways involved in basophil adhesion and histamine release

    DEFF Research Database (Denmark)

    Sha, Quan; Poulsen, Lars K.; Gerwien, Jens

    2006-01-01

    Little is known about basophil with respect to the different signaling transduction pathways involved in spontaneous, cytokine or anti-IgE induced adhesion and how this compares to IgE-dependent and IgE-independent mediator secretion. The purpose of the present study was to investigate the roles ...... of beta1 and beta2 integrins in basophil adhesion as well as hosphatidylinositol 3-kinase (PI3K), src-kinases and extracellular signal regulated kinase (ERK) 1/2 in basophil adhesion and histamine release (HR)....

  4. Signaling transduction pathways involved in basophil adhesion and histamine release

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Background Little is known about basophil with respect to the different signaling transduction pathways involved in spontaneous, cytokine or anti-IgE induced adhesion and how this compares to IgE-dependent and IgE-independent mediator secretion. The purpose of the present study was to investigate the roles of β1 andβ2 integrins in basophil adhesion as well as hosphatidylinositol 3-kinase (PI3K), src-kinases and extracellular signal regulated kinase (ERK)1/2 in basophil adhesion and histamine release (HR). Methods Basophils (purity of 10%-50%) were preincubated with anti-CD29 or anti-CD18 blocking antibodies before used for adhesion study. Basophils were preincubated with the pharmacological inhibitors wortmannin, PP1, PD98059 before used for adhesion and HR study. Cell adherence to bovine serum albumin (BSA) or fibronectin (Fn) was monitored using cell associated histamine as a basophil marker and the histamine was measured by the glass fiber assay.Results Basophil spontaneous adhesion to Fn was inhibited by anti-CD29. Interleukin (IL)-3, granulocyte/macrophage colony stimulating factor (GM-CSF) induced adhesion to BSA was inhibited by anti-CD18. Wortmannin at 1 μmol/L and PP1 at 20 μmol/L strongly interfered with, whereas PD98059 at 50 μmol/L weakly inhibited basophil spontaneous adhesion to Fn. One μmol/L wortmannin strongly inhibited IL-3, IL-5, GM-CSF and anti-IgE induced adhesion to BSA. PP1 at 20 μmol/L partly inhibited anti-IgE induced adhesion. Fifty μmol/L PD98059 marginally inhibited IL-5, weakly inhibited anti-IgE, partly inhibited GM-CSF induced adhesion. Wortmannin, PP1 and PD98059 inhibited anti-IgE (1:100 or 1:1000) induced basophil HR in a dose dependent manner. They inhibited calcium ionophore A23187 (10 μmol/L, 5 μmol/L) induced basophil HR in a dose dependent manner, but to different extend with PP1 being the most efficient.Conclusions Basophil spontaneous adhesion to Fn is mediated by β1-integrins whereas cytokine induced adhesion

  5. The expression of CD123 can decrease with basophil activation: implications for the gating strategy of the basophil activation test

    OpenAIRE

    Santos, Alexandra F.; Bécares, Natalia; Stephens, Alick; Turcanu, Victor; Lack, Gideon

    2016-01-01

    Background Basophil activation test (BAT) reproduces IgE-mediated allergic reactions in vitro and has been used as a diagnostic test. Different markers can be used to identify basophils in whole blood and have implications for the outcome of the test. We aimed to assess changes in the expression of CD123 and HLA-DR following basophil activation and to select the best gating strategy for BAT using these markers. Methods BAT was performed in whole blood from 116 children. Peanut extract, anti-I...

  6. [Human basophil degranulation test. Results of a modified technic (I)].

    Science.gov (United States)

    Román Pérez, F; Berlanga Cortés, J A; Urquiă, M; Guerra Pasadas, F; Peña Martínez, J

    1986-01-01

    The degranulation of human basophils towards antigenic substances, to which they must be previously sensitized as a result of the antigen-antibody union at the membrane level, is a phenomenon that has an important application complementing diagnosis of diseases mediated by the Gell and Coombs classification of Type I Hypersensitivity, on which the principle of Human Basophils Degranulation Test (HBDT) was based. An essential requirement is to be able to collect an adequate quantity of basophils, as well as the visualization by means of a technique of convenient stain. In this work, we show a modified HBDT technique for basophil granulocytes staining. A total of 33 HBDT was carried out on patients sensitive to grass pollen and/or Olea europea pollen and/or D. pteronyssinus, and as controls, HBDT was done on healthy individuals with negative clinical history. Basophil enrichemnt from the circulating blood sample was done by means of centrifugation in Percoll density gradients of 1072, 1076, 1080 and 1082, and we obtained a higher number of basophils with a density of 1080 at 400 Xg for 30 minutes. The cellular bottom was resuspended in 600 microliters of PBS. We employed lyophilized antigenic extracts from grass pollen, Olea europea and D. pteronyssinus, employing weight/volume concentration of 10(-4), 10(-5), 10(-6) and 10(-7). The optimum dilutions were 10(-5) for grass pollen and D. pteronyssinus.

  7. Diagnosis of clavulanic acid allergy using basophil activation and leukotriene release by basophils.

    Science.gov (United States)

    Longo, N; Gamboa, P M; Gastaminza, G; Audícana, M T; Antepara, I; Jaúregui, I; Sanz, M L

    2008-01-01

    Clavulanic acid is a potent inhibitor of B-lactamase that is increasingly prescribed in association with amoxicillin. We report 2 cases of patients who experienced pruritus, wheals, and angioedema after oral intake of amoxicillin/clavulanic acid. Routine skin tests for B-lactam antibiotics and specific immunoglobulin (Ig) E were negative in both patients. Analysis of CD63 expression by the basophil activation test (BAT) using flow cytometry and of sulphidoleukotriene (sLT) release by basophils using the cellular allergen stimulation test (CAST) revealed significant positive responses with amoxicillin/clavulanic acid and with clavulanic acid, and negative responses with amoxicillin and other beta-lactam antibiotics. In addition, cultured CD3+CD4+ cells showed a significant increase in the expression of CD69, CD25, and HLA-DR in the presence of clavulanic acid. Both patients tolerated therapeutic doses of amoxicillin. BAT and CAST are useful ex vivo procedures for the detection of specific IgE-mediated allergy to clavulanic acid, especially for patients with negative skin test results.

  8. Basophil degranulation induced by oral poison ivy antigen.

    Science.gov (United States)

    Shelley, W B; Resnik, S S

    1965-08-01

    Seven subjects shown by patch test to be sensitive to poison ivy oleoresin were challenged with graded oral doses of ivy extract. In each instance the circulating basophil leukocytes showed significant degranulation within one hour of challenge. This finding was interpreted as evidence of the presence of immediate-type circulating antibody to ivy antigen in these subjects. No drop in the absolute basophil count was noted, but with higher oral doses the degranulation persisted for several days. Thirteen control subjects showed no change in the basophil morphology or count, indicating that the resin at these levels was not toxic to this cell. All but one of the sensitive subjects showed objective patch test evidence of hyposensitization following the intensive three-week course of oral poison ivy antigen.

  9. Identification and quantification of basophils in the airways of asthmatics following segmental allergen challenge.

    Science.gov (United States)

    Dijkstra, Dorothea; Hennig, Christian; Hansen, Gesine; Biller, Heike; Krug, Norbert; Hohlfeld, Jens M

    2014-07-01

    During asthma attacks, allergens activate sensitized basophils in the lung, thereby aggravating symptoms. Due to the paucity of basophils in bronchial lavage fluid and the lack of specific basophil detection and quantification methods, basophil-directed research in these samples was hampered in the past. This study aimed to establish and validate a flow cytometry-based basophil detection and quantification method for human basophils from bronchoalveolar lavage (BAL) and blood as a prerequisite for a better understanding of their pathogenic contribution and subtyping of asthma phenotypes. BAL and blood leukocytes from seasonal asthmatics were analyzed by flow cytometry. Chipcytometry, a highly sensitive single-cell analysis method, was used to validate the staining panel for basophils. Cell differentials of May-Grünwald-Giemsa-stained cytospins were used to compare basophil percentages. BAL basophils are identifiable as CD123(+) HLA-DR(-) CD3(-) CD14(-) CD19(-) CD20(-) CD56(-) cells in flow cytometrical analysis. Their identity was validated by Chipcytometry. CD203c was highly expressed by BAL basophils, whereas it was expressed at variable levels on blood basophils. The two quantification methods correlated, although more basophils were detected by flow cytometry. Furthermore, the increase in basophil percentages in the lung correlated with the decrease in the basophil percentages in the blood after allergen challenge. We here validated a reliable basophil quantification method, which is independent of the cell's activation and degranulation state. The results obtained with this method indicate that basophils are directly recruited from the blood circulation to the airway lumen.

  10. Orientation de la réponse immune par les basophiles

    OpenAIRE

    Galeotti, C.; Stephen-Victor, E.; Sharma, M; Kaveri, S.V.; Bayry, J.

    2016-01-01

    International audience; Basophils are infrequent granulocytes which are important for Th2 responses, protection against helminthic parasites and the pathogenesis of allergy and other inflammatory conditions. Recent reports suggest that in mice basophils function as antigen-presenting cells for mediation of Th2 responses. However, human basophils lack features of antigen-presenting cells.; Les basophiles sont de rares granulocytes et ont un rôle important dans les réponses Th2, dans la protect...

  11. Upregulated expression of substance P in basophils of the patients with chronic spontaneous urticaria: induction of histamine release and basophil accumulation by substance P.

    Science.gov (United States)

    Zheng, Wenjiao; Wang, Junling; Zhu, Wei; Xu, Chiyan; He, Shaoheng

    2016-06-01

    Human basophils have been implicated in the pathogenesis of chronic spontaneous urticaria (CSU), and substance P (SP) is a possible candidate as histamine-releasing factor in some patients with CSU. However, little is known of relationship between basophils and SP in CSU. In the present study, we investigated expression of SP and NK1R on basophils from patients with CSU, and influence of SP on basophil functions by using flow cytometry analysis, basophil challenge, and mouse sensitization model techniques. The results showed that plasma SP level and basophil numbers in CSU patients were higher than that in HC subject. The percentages of SP+ and NK1R+ basophils were markedly elevated in CSU blood in comparison with HC blood. Once added, SP induced up to 41.2 % net histamine release from basophils of CSU patients, which was comparable with that provoked by anti-IgE, and fMLP. It appeared that SP induced dramatic increase in blood basophil numbers of mice following peritoneal injection. Ovalbumin (OVA)-sensitized mice had much more SP+ and NK1R+ basophils in blood than non-sensitized mice. In conclusion, the elevated plasma concentration of SP, upregulated expression of SP and NK1R on basophils, and the ability of SP in induction of basophil degranulation and accumulation indicate strongly that SP is most likely a potent proinflammatory mediator, which contributes greatly to the pathogenesis of CSU through basophils. Inhibitors of SP and blockers of NK1R are likely useful agents for treatment of CSU.

  12. Evaluation of Basophil Infiltration into the Skin Lesions of Tick Bites

    Directory of Open Access Journals (Sweden)

    Kyoko Nakahigashi

    2013-02-01

    Full Text Available Recently, it has been described that basophils play an essential role in antibody-mediated acquired immunity against ticks in mice. However, it is still unknown whether basophil infiltration has any significance in the infestation with ticks in humans. In this report, we have evaluated the infiltration of basophils into human skin lesions of tick bites.

  13. Luteolin, a flavonoid, inhibits AP-1 activation by basophils.

    Science.gov (United States)

    Hirano, Toru; Higa, Shinji; Arimitsu, Junsuke; Naka, Tetsuji; Ogata, Atsushi; Shima, Yoshihito; Fujimoto, Minoru; Yamadori, Tomoki; Ohkawara, Tomoharu; Kuwabara, Yusuke; Kawai, Mari; Matsuda, Hisashi; Yoshikawa, Masayuki; Maezaki, Naoyoshi; Tanaka, Tetsuaki; Kawase, Ichiro; Tanaka, Toshio

    2006-02-03

    Flavonoids including luteolin, apigenin, and fisetin are inhibitors of IL-4 synthesis and CD40 ligand expression by basophils. This study was done to search for compounds with greater inhibitory activity of IL-4 expression and to clarify the molecular mechanisms through which flavonoids inhibit their expression. Of the 37 flavonoids and related compounds examined, ayanin, luteolin, and apigenin were the strongest inhibitors of IL-4 production by purified basophils in response to anti-IgE antibody plus IL-3. Luteolin did not suppress Syk or Lyn phosphorylation in basophils, nor did suppress p54/46 SAPK/JNK, p38 MAPK, and p44/42 MAPK activation by a basophilic cell line, KU812 cells, stimulated with A23187 and PMA. However, luteolin did inhibit phosphorylation of c-Jun and DNA binding activity of AP-1 in nuclear lysates from stimulated KU812 cells. These results provide a fundamental structure of flavonoids for IL-4 inhibition and demonstrate a novel action of flavonoids that suppresses the activation of AP-1.

  14. Spontaneous and cytokine induced basophil adhesion evaluated by microtiter assay

    DEFF Research Database (Denmark)

    Quan, Sha; Poulsen, Lars K; Reimert, Claus Michael

    2002-01-01

    by the histamine content of the adhering basophils. The spontaneous adhesion to fibronectin was higher than to laminin and collagen type I. Both spontaneous adhesion to fibronectin and interleukin-3 (IL-3), interleukin-5 (IL-5), granulocyte/macrophage colony stimulating factor (GM-CSF) induced adhesion to BSA...

  15. Importance of basophil activation testing in insect venom allergy

    Directory of Open Access Journals (Sweden)

    Kosnik Mitja

    2009-12-01

    Full Text Available Abstract Background Venom immunotherapy (VIT is the only effective treatment for prevention of serious allergic reactions to bee and wasp stings in sensitized individuals. However, there are still many questions and controversies regarding immunotherapy, like selection of the appropriate allergen, safety and long term efficacy. Methods Literature review was performed to address the role of basophil activation test (BAT in diagnosis of venom allergy. Results In patients with positive skin tests or specific IgE to both honeybee and wasp venom, IgE inhibition test can identify sensitizing allergen only in around 15% and basophil activation test increases the identification rate to around one third of double positive patients. BAT is also diagnostic in majority of patients with systemic reactions after insect stings and no detectable IgE. High basophil sensitivity to allergen is associated with a risk of side effects during VIT. Persistence of high basophil sensitivity also predicts a treatment failure of VIT. Conclusion BAT is a useful tool for better selection of allergen for immunotherapy, for identification of patients prone to side effects and patients who might be treatment failures. However, long term studies are needed to evaluate the accuracy of the test.

  16. Bacterial flagellin induces IL-6 expression in human basophils.

    Science.gov (United States)

    Jeon, Jun Ho; Ahn, Ki Bum; Kim, Sun Kyung; Im, Jintaek; Yun, Cheol-Heui; Han, Seung Hyun

    2015-05-01

    Binding of allergen to IgE on basophils positively affects allergic inflammation by releasing inflammatory mediators. Recently, basophils were shown to express pattern-recognition receptors, such as toll-like receptors (TLRs), for recognizing microbe-associated molecular patterns (MAMPs) that are independent of allergen-IgE binding. In this study, we investigated whether MAMP alone can induce IL-6 production in a human basophil cell line, KU812. Stimulation with flagellin in the absence of allergen-IgE association induced IL-6 expression in KU812 cells, while stimulation with lipoteichoic acid, peptidoglycan, or poly I:C did not under the same condition. Flagellin-induced IL-6 expression was also observed in human primary basophils. Flow cytometric analysis showed that KU812 cells expressed flagellin-recognizing TLR5 both on the cell surface and in the cytoplasm while TLR2 and TLR3 were observed only in the cytoplasm. We further demonstrated that although flagellin augmented the phosphorylation of mitogen-activated protein kinases including p38 kinase, ERK, and JNK, flagellin-induced IL-6 production was attenuated by inhibitors for p38 kinase and ERK, but not by JNK inhibitors. In addition, flagellin enhanced phosphorylation of signaling molecules including CREB, PKCδ, and AKT. The inhibitors for PKA and PKC also showed inhibitory effects. Interestingly, flagellin-induced IL-6 production was further enhanced by pretreatment with inhibitors for PI3K, implying that PI3K negatively affects the flagellin-induced IL-6 production. Furthermore, DNA binding activities of NF-κB, AP-1, and CREB, which play pivotal roles in the induction of IL-6 gene expression, were increased by flagellin. These results suggest that flagellin alone is sufficient to induce IL-6 gene expression via TLR5 signaling pathways in human basophils.

  17. Basophil-derived IL-6 regulates TH17 cell differentiation and CD4 T cell immunity

    Science.gov (United States)

    Yuk, Chae Min; Park, Hyeung Ju; Kwon, Bo-In; Lah, Sang Joon; Chang, Jun; Kim, Ji-Young; Lee, Kyung-Mi; Park, Su-Hyung; Hong, Seokchan; Lee, Seung-Hyo

    2017-01-01

    Basophils are rare, circulating granulocytes proposed to be involved in T helper (TH) type 2 immunity, mainly through secretion of interleukin (IL)-4. In addition to IL-4, basophils produce IL-6 and tumor necrosis factor (TNF)-α in response to immunoglobulin E (IgE) crosslinking. Differentiation of TH17 cells requires IL-6 and transforming growth factor (TGF)-β, but whether basophils play a significant role in TH17 induction is unknown. Here we show a role for basophils in TH17 cell development by using in vitro T cell differentiation and in vivo TH17-mediated inflammation models. Bone marrow derived-basophils (BMBs) and splenic basophils produce significant amounts of IL-6 as well as IL-4 following stimulation with IgE crosslink or cholera toxin (CT). In addition, through IL-6 secretion, BMBs cooperate with dendritic cells to promote TH17 cell differentiation. In the TH17 lung inflammation model, basophils are recruited to the inflamed lungs following CT challenge, and TH17 responses are significantly reduced in the absence of basophils or IL-6. Furthermore, reconstitution with wild-type, but not IL-6-deficient, basophils restored CT-mediated lung inflammation. Lastly, basophil-deficient mice showed reduced phenotypes of TH17-dependent experimental autoimmune encephalomyelitis. Therefore, our results indicate that basophils are an important inducer of TH17 cell differentiation, which is dependent on IL-6 secretion. PMID:28134325

  18. An IL-1 cytokine member, IL-33, induces human basophil activation via its ST2 receptor.

    Science.gov (United States)

    Suzukawa, Maho; Iikura, Motoyasu; Koketsu, Rikiya; Nagase, Hiroyuki; Tamura, Chise; Komiya, Akiko; Nakae, Susumu; Matsushima, Kouji; Ohta, Ken; Yamamoto, Kazuhiko; Yamaguchi, Masao

    2008-11-01

    Basophils are thought to play pivotal roles in allergic inflammation through rapid release of chemical mediators in addition to sustained production of Th2 cytokines, including IL-4. A newly identified cytokine, IL-33, has been recognized as one of the key cytokines enhancing Th2-balanced immune regulation through its receptor, ST2. The present study was conducted to elucidate whether IL-33 acts directly on, and affects the functions of, human basophils. Real-time PCR analysis showed that basophils express transcripts for ST2. The expression levels were significantly higher compared with eosinophils and neutrophils, and treatment with IL-33 significantly up-regulated basophil ST2 mRNA expression. Expressions of IL-4 and IL-13 mRNA were also up-regulated by IL-33, and there was also enhanced secretion of IL-4 protein. IL-33 increased the surface levels of basophil CD11b expression and enhanced basophil adhesiveness. Although IL-33 failed to directly induce degranulation or attract basophils, it exerted priming effects on basophils. It enhanced degranulation in response to IgE-crosslinking stimulus and also enhanced basophil migration toward eotaxin without changing surface CCR3. Also, IL-33 synergistically enhanced IL-4 production and CD11b expression by IL-3-stimulated basophils. Neutralization using Ab specific for ST2 significantly diminished the enhancing effects of IL-33 on both basophil CD11b expression and migration toward eotaxin, indicating that IL-33 signals via ST2 expressed on basophils. This study revealed that IL-33 potently regulates migration and activation of human basophils. IL-33 may be a key cytokine in the pathogenesis of Th2-dominant inflammation by acting not only on lymphocytes but also on effector cells such as basophils.

  19. Comparative electron microscopy of basophils and mast cells, in vivo and in vitro.

    Science.gov (United States)

    Eguchi, M

    1991-01-01

    We compared the fine structure and electron microscopic cytochemical findings of basophils and mast cells from humans, guinea pigs, rabbits, mice and rats. The particulate structure was the most frequently observed and most typical structure of human and rabbit basophil granules and of guinea pig mast cell granules. The most prominent feature of guinea pig basophils and murine mast cells was that the fine structure of the granules was homogeneous. The fine structure of the granules in guinea pig basophils resembled that in murine mast cells, while the fine structure of the granules of guinea pig mast cells resembled those in human and rabbit mast cells. In mouse mast cells in culture, the majority of the granules contained small vesicles, which were also observed in human basophils in culture and in mouse basophils in vivo. The degrees of cytochemical reactivity of acid mucopolysaccharides among the species were different. Peroxidase activity was positive in most basophils and in human mast cells. Among mammals, the granules of basophils and mast cells present heterogeneous fine structure. It is of interest that the basophil granules of some species resemble the mast cell granules rather than the basophil granules of other species.

  20. Role of human mast cells and basophils in bronchial asthma.

    Science.gov (United States)

    Marone, Gianni; Triggiani, Massimo; Genovese, Arturo; De Paulis, Amato

    2005-01-01

    Mast cells and basophils are the only cells expressing the tetrameric (alphabetagamma2) structure of the high affinity receptor for IgE (FcepsilonRI) and synthesizing histamine in humans. Human FcepsilonRI+ cells are conventionally considered primary effector cells of bronchial asthma. There is now compelling evidence that these cells differ immunologically, biochemically, and pharmacologically, which suggests that they might play distinct roles in the appearance and fluctuation of the asthma phenotype. Recent data have revealed the complexity of the involvement of human mast cells and basophils in asthma and have shed light on the control of recruitment and activation of these cells in different lung compartments. Preliminary evidence suggests that these cells might not always be detrimental in asthma but, under some circumstances, they might exert a protective effect by modulating certain aspects of innate and acquired immunity and allergic inflammation.

  1. Development and characterization of a monoclonal antibody specific for human basophils and the identification of a unique secretory product of basophil activation.

    Science.gov (United States)

    McEuen, A R; Buckley, M G; Compton, S J; Walls, A F

    1999-01-01

    Despite increasing evidence that basophils can infiltrate into inflamed tissues during allergic reactions, determination of the extent of infiltration and elucidation of their role in allergic disease has been frustrated by the lack of reliable means for detecting this cell type in tissues. In the present study, we report on a new monoclonal antibody specific for basophils and on the initial characterization of the antigen it recognizes. Basophils were isolated from peripheral blood by Percoll density gradient centrifugation and a positive-selection immunomagnetic procedure and injected into mice to produce monoclonal antibodies. A hybridoma clone, designated BB1, secreted antibody of the IgG2a isotype; this antibody bound selectively to basophils on immunocytochemistry but did not react with any other cell type or tissue structure, although it did stain a proportion of cells from the basophilic cell line KU812F. In sections of mixed populations of peripheral blood cells, similar numbers of cells stained with Alcian blue dye and BB1 over a wide range of basophil purity. BB1 antibody was effective in identifying basophils in sections of mixed cells or in tissues after fixation with ethanol, Carnoy's solution, or formalin. Staining of basophils with BB1 gave a granular appearance, although flow cytometry indicated that some antigen was also present on the surface of the cell. Activation of these cells with anti-IgE antibody or with the calcium ionophore A23187 provoked release of the antigen in parallel with that of histamine. BB1 antibody did not, by itself, stimulate histamine release. The molecular mass of the antigen was determined on Hedrick-Smith gels to be 124+/-11 kd. This new monoclonal antibody will be a valuable experimental tool in future studies, allowing the reliable detection of basophils in tissues of patients with allergic and chronic inflammatory disease; in addition, the antigen it identifies has potential as a unique marker of basophil activation.

  2. Histamine content and secretion in basophils and mast cells.

    Science.gov (United States)

    Dvorak, A M

    1998-01-01

    Biochemical determinations of the histamine content and secretion from basophils and mast cells have been available for some time, and much of the complex anatomy of these cellular populations and their release reactions has been documented using the electron microscope. The ultrastructural analyses led to the description of vesicular transport between secretory granules and the plasma membrane as a mechanism for secretion from basophils and mast cells--a process termed piecemeal degranulation. Proof of concepts incorporated in a general degranulation model put forth in 1975 (DVORAK, H.F. and DVORAK, A.M.) requires high magnification imaging of a granule constituent in trafficking vesicles in the process of a stimulated release reaction in which the constituent release is monitored biochemically. Development and application of a new enzyme-affinity method to detect histamine at high magnifications in well-preserved ultrastructural samples have provided the necessary means to establish proof that appropriate secretagogues can stimulate the vesicular transport of histamine in basophils and mast cells during release reactions monitored biochemically. The background information necessary to the understanding of this result is presented here, as well as the development and verification of the diamine oxidase-gold method to image histamine in human mast cell granules as the test system. Also presented are applications using this technology to examine histamine stores and secretion in vitro, in vivo, and ex vivo in human basophils and mast cells and in mouse mast cells. Specifically examined are histamine stores developing in maturing mast cells induced to develop de novo from cultured human cord blood cells, secretagogue-stimulated release and recovery of histamine stores from isolated, purified human lung mast cells ex vivo, cytokine-stimulated degranulation of human skin mast cells and their histamine stores in vivo, piecemeal degranulation of human gut mast cells and

  3. 89 Is Basophil Specific Response to Hymenoptera Venom Related to T Regulatory Cells?

    OpenAIRE

    Kucera, Petr; Hulikova, Katarina; Cvackova, Milada; Planska, Daniela; Riegerova, Kamila

    2012-01-01

    Background The exact mechanism of systemic hypersensitivity to venom is not exactly understood. It is suggested T cells with regulatory potential can downregulate other T cell subsets and effector cells, ex. mast cell or basophils. We focused on relationship of specific basophil reactivity in relationship to proportion of regulatory T cells. Methods Forty-five patients with history of systemic symptoms of allergy to Hymenoptera venom were included. Basophil reactivity before the treatment and...

  4. Development of a microarray-based method to detect exposure of human basophils to IL-3.

    Science.gov (United States)

    MacGlashan, Donald

    2012-11-30

    Human basophils are an accessible participant of the human allergic reaction. There is natural variation in various functional endpoints and in signaling molecule expression but there has been only a limited effort to place this information in the context of mRNA expression profiles. The goal of these studies was to develop gene transcript profiles associated with IL-3 exposure for use in probing basophil phenotypes. Highly purified human basophils were cultured under several conditions and the cells subsequently analyzed by microarray analysis of gene transcript expression. Basophils were cultured with or without IL-3 for 24 h and the expression profile, relative to pre-culture, analyzed for transcripts that act in a reciprocal fashion depending on the condition. A 51-gene transcript set was identified that was composed of 34 transcripts that increased with IL-3 and decreased without IL-3 and a further 17 transcripts that did the reverse. This set of transcripts was validated on two microarray chips. The changes induced by IL-3 were stable in time, with 3 day cultures generating a signature concordant with 1 day cultures. The inverted nature of the response to ±IL-3 suggested that the normal circulating basophil is balanced between a state of high and low IL-3 exposure and thus is very sensitive to changes in IL-3. For example, the basophil could detect the early generation of IL-3 that follows IgE-mediated stimulation of basophils. When this signature tool was applied to freshly isolated basophils, it was observed that the signature was similar for the same subjects' basophil sampled weeks apart. It was also shown that the 51-gene transcript was insensitive to the method of preparing purified basophils. Finally, these studies provided an estimate for the normal in vivo exposure of circulating basophils to IL-3 or IL-3-like functionality. These studies identified an IL-3 signature to probe changes in basophils occurring in vivo.

  5. Short-, Intermediate-, and Long-Term Changes in Basophil Reactivity Induced by Venom Immunotherapy

    OpenAIRE

    Rodríguez Trabado, Ana; Cámara Hijón, Carmen; Ramos Cantariño, Alfonso; Romero-Chala, Silvia; García-Trujillo, José Antonio; Fernández Pereira, Luis Miguel

    2016-01-01

    Purpose The basophil activation test (BAT) has been used to monitor venom immunotherapy (VIT) due to its high specificity. A previous study has reported a good correlation between a significant decrease in basophil activation during 5 years of VIT and clinical protection assessed by sting challenge. The following prospective study was performed to examine changes in basophil reactivity over a complete VIT period of 5 years. Methods BAT in a dose-response curve was studied prospectively in 10 ...

  6. Contribution of basophils to cutaneous immune reactions and Th2-mediated allergic responses

    Directory of Open Access Journals (Sweden)

    Atsushi eOtsuka

    2015-08-01

    Full Text Available Basophils are potent effector cells of innate immunity and also play a role in T helper 2 (Th2-mediated allergic responses. But, although their in vitro functions are well studied, their in vivo functions remain largely unknown. However, several mouse models of basophil depletion have recently been developed and used to investigate basophil functions. For example, in a croton oil-induced model of irritant contact dermatitis in conditionally basophil-depleted transgenic mice, we found that basophils rapidly infiltrate inflamed skin and subsequently induce infiltration of eosinophils. We also showed that basophils induce Th2 skewing upon epicutaneous sensitization with various haptens and peptide antigens. Intriguingly, basophils also promoted Th2 polarization upon protein antigen exposure in the presence of dendritic cells (DCs. The dermal DC subset associated with Th2 skewing was recently identified as CD301b+ DC. Such studies with basophil-deficient mouse models have significantly improved our understanding of the mechanisms involved in human immune-related diseases. In this review, we will focus on the relative contribution of basophils and DCs to Th2-mediated allergic responses.

  7. Evaluation of Allergy Effector Cell Function: Suppression of Basophils in Chronic Helminth Infections

    Science.gov (United States)

    2011-01-01

    basophils are probably best known for their participation in the effector phase of immediate hypersensitivity reactions, some studies have suggested...had Schistosoma mansoni infections were then sensitized to Penicillin V. When mice in these groups were challenged with Pen V, body temperature was... hypersensitivity reactions, reduced basophil functionality could be one way helminth infection protects against allergic disease. 2. Determine the

  8. Increased level of basophil CD203c expression predicts severe chronic urticaria.

    Science.gov (United States)

    Ye, Young-Min; Yang, Eun-Mi; Yoo, Hye-Soo; Shin, Yoo-Seob; Kim, Seung-Hyun; Park, Hae-Sim

    2014-01-01

    Increased FcεR1α expression with upregulated CD203c expression on peripheral basophils is seen in patients with chronic urticaria (CU). However, there has been no published report on the association between CD203c expression level and clinical disease activity in CU patients. To investigate whether the increase of basophil activation is associated with the disease activity of CU, we measured basophil CD203c expression using a tricolor flow cytometric method in 82 CU patients and 21 normal controls. The relationship between the percentage of CD203c-expressing basophils and clinical parameters was analyzed. The mean basophil CD203c expression was significantly higher in CU patients than in healthy controls (57.5% vs 11.6%, P basophil CD203c expression in severe CU patients was significantly higher than in non-severe CU (66.5% ± 23.3% vs 54.0% ± 23.3%, P = 0.033). Multiple logistic regression analysis indicated that both ≥ 72% basophil CD203c expression and urticaria activity score (UAS)≥ 13 were significant predictors of severe CU (P = 0.005 and P = 0.032, respectively). These findings suggest that the quantification of basophil activation with CD203c at baseline may be used as a potential predictor of severe CU requiring another treatment option beyond antihistamines.

  9. Unusual cause of sudden cardiac death : Basophilic degeneration of coronary arteries

    NARCIS (Netherlands)

    Jessurun, GAJ; Tio, RA; Ribbert, LSM; Willemse, F; Boonstra, PW; Crijns, HJGM

    1996-01-01

    Basophilic or mucoid degeneration of the intima in the coronary arteries is an extremely rare cause of premature atherosclerosis. An unusual case of fatal basophilic degeneration of the coronary arteries 142 days after delivery is reported. (C) 1996 Wiley-Liss, Inc.

  10. Cysteine protease antigens cleave CD123, the α subunit of murine IL-3 receptor, on basophils and suppress IL-3-mediated basophil expansion

    Energy Technology Data Exchange (ETDEWEB)

    Nishikado, Hideto [Atopy (Allergy) Research Center, Juntendo University Graduate School of Medicine, Tokyo (Japan); Fujimura, Tsutomu; Taka, Hikari; Mineki, Reiko [Laboratory of Proteomics and Biomolecular Science, BioMedical Research Center, Juntendo University Graduate School of Medicine, Tokyo (Japan); Ogawa, Hideoki; Okumura, Ko [Atopy (Allergy) Research Center, Juntendo University Graduate School of Medicine, Tokyo (Japan); Takai, Toshiro, E-mail: t-takai@juntendo.ac.jp [Atopy (Allergy) Research Center, Juntendo University Graduate School of Medicine, Tokyo (Japan)

    2015-05-01

    Th2 type immune responses are essential for protective immunity against parasites and play crucial roles in allergic disorders. Helminth parasites secrete a variety of proteases for their infectious cycles including for host entry, tissue migration, and suppression of host immune effector cell function. Furthermore, a number of pathogen-derived antigens, as well as allergens such as papain, belong to the family of cysteine proteases. Although the link between protease activity and Th2 type immunity is well documented, the mechanisms by which proteases regulate host immune responses are largely unknown. Here, we demonstrate that the cysteine proteases papain and bromelain selectively cleave the α subunit of the IL-3 receptor (IL-3Rα/CD123) on the surface of murine basophils. The decrease in CD123 expression on the cell surface, and the degradation of the extracellular domain of recombinant CD123 were dependent on the protease activity of papain and bromelain. Pre-treatment of murine basophils with papain resulted in inhibition of IL-3-IL-3R signaling and suppressed IL-3- but not thymic stromal lymphopoietin-induced expansion of basophils in vitro. Our unexpected findings illuminate a novel mechanism for the regulation of basophil functions by protease antigens. Because IL-3 plays pivotal roles in the activation and proliferation of basophils and in protective immunity against helminth parasites, pathogen-derived proteases might contribute to the pathogenesis of infections by regulating IL-3-mediated functions in basophils. - Highlights: • We identified the murine IL3R as a novel target of papain-family cysteine proteases. • Papain-family cysteine proteases cleaved IL3Rα/CD123 on murine basophils. • Papain suppressed IL3- but not TSLP-induced expansion of murine basophils. • The inactivation of IL3R might be a strategy for pathogens to suppress host immunity.

  11. Basophil activation test with food additives in chronic urticaria patients.

    Science.gov (United States)

    Kang, Min-Gyu; Song, Woo-Jung; Park, Han-Ki; Lim, Kyung-Hwan; Kim, Su-Jung; Lee, Suh-Young; Kim, Sae-Hoon; Cho, Sang-Heon; Min, Kyung-Up; Chang, Yoon-Seok

    2014-01-01

    The role of food additives in chronic urticaria (CU) is still under investigation. In this study, we aimed to explore the association between food additives and CU by using the basophil activation test (BAT). The BAT using 15 common food additives was performed for 15 patients with CU who had a history of recurrent urticarial aggravation following intake of various foods without a definite food-specific IgE. Of the 15 patients studied, two (13.3%) showed positive BAT results for one of the tested food additives. One patient responded to monosodium glutamate, showing 18.7% of CD203c-positive basophils. Another patient showed a positive BAT result to sodium benzoate. Both patients had clinical correlations with the agents, which were partly determined by elimination diets. The present study suggested that at least a small proportion of patients with CU had symptoms associated with food additives. The results may suggest the potential utility of the BAT to identity the role of food additives in CU.

  12. The STAT5-GATA2 pathway is critical in basophil and mast cell differentiation and maintenance.

    Science.gov (United States)

    Li, Yapeng; Qi, Xiaopeng; Liu, Bing; Huang, Hua

    2015-05-01

    Transcription factor GATA binding protein 2 (GATA2) plays critical roles in hematopoietic stem cell survival and proliferation, granulocyte-monocyte progenitor differentiation, and basophil and mast cell differentiation. However, precise roles of GATA2 in basophil and mast cell differentiation and maintenance have not been delineated. We have identified GATA2 as an essential transcription factor in differentiation of newly identified common basophil and mast cell progenitors into basophils and mast cells. We observed Gata2 haploinsufficiency for mast cell differentiation, but not for basophil differentiation. We examined the precise role of GATA2 in maintaining the expression of a wide range of genes that are important for performing basophil or mast cell functions. The effects of GATA2 on gene expression were broadly based. We demonstrated that GATA2 was required for maintaining Fcer1a mRNA and FcεRIα protein expression on both basophils and mast cells, as well as for maintaining Kit mRNA and c-Kit protein expression on mast cells. GATA2 was required for histamine synthesis and was also critical for Il4 mRNA expression in basophils and Il13 mRNA expression in mast cells. We demonstrate a STAT5-GATA2 connection, showing that the STAT5 transcription factor directly bound to the promoter and an intronic region of the Gata2 gene. Overexpression of the Gata2 gene was sufficient to direct basophil and mast cell differentiation in the absence of the Stat5 gene. Our study reveals that the STAT5-GATA2 pathway is critical for basophil and mast cell differentiation and maintenance.

  13. Mouse basophils reside in extracellular matrix-enriched bone marrow niches which control their motility.

    Science.gov (United States)

    Smaniotto, Salete; Schneider, Elke; Goudin, Nicolas; Bricard-Rignault, Rachel; Machavoine, François; Dardenne, Mireille; Dy, Michel; Savino, Wilson

    2013-01-01

    Basophils co-express FcεRIα and CD49b, the α-2 chain of integrin-type receptor VLA-2 (α2β1), which recognizes type-1 collagen as a major natural ligand. The physiological relevance of this integrin for interactions with extracellular bone marrow matrix remains unknown. Herein, we examined the expression of several receptors of this family by bone marrow-derived basophils sorted either ex-vivo or after culture with IL-3. Having established that both populations display CD49d, CD49e and CD49f (α-4, α-5 and α-6 integrins subunits, respectively), we addressed receptor functions by measuring migration, adhesion, proliferation and survival after interacting with matched natural ligands. Type I collagen, laminin and fibronectin promoted basophil migration/adhesion, the former being the most effective. None of these ligands affected basophil viability and expansion. Interactions between basophils and extracellular matrix are likely to play a role in situ, as supported by confocal 3D cell imaging of femoral bone marrow sections, which revealed basophils exclusively in type-1 collagen-enriched niches that contained likewise laminin and fibronectin. This is the first evidence for a structure/function relationship between basophils and extracellular matrix proteins inside the mouse bone marrow.

  14. Mouse basophils reside in extracellular matrix-enriched bone marrow niches which control their motility.

    Directory of Open Access Journals (Sweden)

    Salete Smaniotto

    Full Text Available Basophils co-express FcεRIα and CD49b, the α-2 chain of integrin-type receptor VLA-2 (α2β1, which recognizes type-1 collagen as a major natural ligand. The physiological relevance of this integrin for interactions with extracellular bone marrow matrix remains unknown. Herein, we examined the expression of several receptors of this family by bone marrow-derived basophils sorted either ex-vivo or after culture with IL-3. Having established that both populations display CD49d, CD49e and CD49f (α-4, α-5 and α-6 integrins subunits, respectively, we addressed receptor functions by measuring migration, adhesion, proliferation and survival after interacting with matched natural ligands. Type I collagen, laminin and fibronectin promoted basophil migration/adhesion, the former being the most effective. None of these ligands affected basophil viability and expansion. Interactions between basophils and extracellular matrix are likely to play a role in situ, as supported by confocal 3D cell imaging of femoral bone marrow sections, which revealed basophils exclusively in type-1 collagen-enriched niches that contained likewise laminin and fibronectin. This is the first evidence for a structure/function relationship between basophils and extracellular matrix proteins inside the mouse bone marrow.

  15. The basophil activation test by flow cytometry: recent developments in clinical studies, standardization and emerging perspectives

    Directory of Open Access Journals (Sweden)

    Debard Anne-Lise

    2005-06-01

    Full Text Available Abstract The diagnosis of immediate allergy is mainly based upon an evocative clinical history, positive skin tests (gold standard and, if available, detection of specific IgE. In some complicated cases, functional in vitro tests are necessary. The general concept of those tests is to mimic in vitro the contact between allergens and circulating basophils. The first approach to basophil functional responses was the histamine release test but this has remained controversial due to insufficient sensitivity and specificity. During recent years an increasing number of studies have demonstrated that flow cytometry is a reliable tool for monitoring basophil activation upon allergen challenge by detecting surface expression of degranulation/activation markers (CD63 or CD203c. This article reviews the recent improvements to the basophil activation test made possible by flow cytometry, focusing on the use of anti-CRTH2/DP2 antibodies for basophil recognition. On the basis of a new triple staining protocol, the basophil activation test has become a standardized tool for in vitro diagnosis of immediate allergy. It is also suitable for pharmacological studies on non-purified human basophils. Multicenter studies are now required for its clinical assessment in large patient populations and to define the cut-off values for clinical decision-making.

  16. The basophil activation test by flow cytometry: recent developments in clinical studies, standardization and emerging perspectives.

    Science.gov (United States)

    Boumiza, Radhia; Debard, Anne-Lise; Monneret, Guillaume

    2005-06-30

    The diagnosis of immediate allergy is mainly based upon an evocative clinical history, positive skin tests (gold standard) and, if available, detection of specific IgE. In some complicated cases, functional in vitro tests are necessary. The general concept of those tests is to mimic in vitro the contact between allergens and circulating basophils. The first approach to basophil functional responses was the histamine release test but this has remained controversial due to insufficient sensitivity and specificity. During recent years an increasing number of studies have demonstrated that flow cytometry is a reliable tool for monitoring basophil activation upon allergen challenge by detecting surface expression of degranulation/activation markers (CD63 or CD203c). This article reviews the recent improvements to the basophil activation test made possible by flow cytometry, focusing on the use of anti-CRTH2/DP2 antibodies for basophil recognition. On the basis of a new triple staining protocol, the basophil activation test has become a standardized tool for in vitro diagnosis of immediate allergy. It is also suitable for pharmacological studies on non-purified human basophils. Multicenter studies are now required for its clinical assessment in large patient populations and to define the cut-off values for clinical decision-making.

  17. The effect of stimuli on basophil-mediated atopic responses during asthmatic lying-in women and in newborns.

    Science.gov (United States)

    Yang, Ling; Guo, Yin-Shi; Jiang, Jin-Qi; Guo, Xue-Jun; Xu, Yi-Ping; Tian, Ye; Xiong, Ying; Han, Li

    2012-08-01

    Morbidity from allergic diseases is increasing. Basophils play a critical role in systemic anaphylaxis and chronic allergic inflammation. The prenatal environment must be regarded as a possible early risk factor for allergic diseases in children. Our objective was to determine if basophils harvested from neonates genetically predisposed to atopic disease had different levels of CD63 expression and IL-4 release properties in response to various stimuli (peptidoglycan, Dermatophagoides farinae, hyperosmotic mannitol). Blood samples were collected from 16 asthmatic and 18 healthy women and their newborns. Peripheral blood basophil histamine was measured using the human basophil degranulation test (HBDT), whereas activation was assessed by flow cytometric measurement of CD63 expression on the cord blood basophil surface. IL-4 levels were quantified by ELISA following allergen stimulation. The basophil degranulation index (DI) in granulocytes harvested from the peripheral blood of asthmatic women was assessed following stimulation with peptidoglycan (PGN), Dermatophagoides farinae (Df ) extract, or hyperosmotic mannitol. The DI was significantly higher in atopic women than in healthy controls. Upregulation of CD63 on the cord blood basophil surface was also detected in response to these stimuli. Basophils purified from the cord blood of neonates born to atopic mothers produced more IL-4 compared to basophils purified from the controls. These data suggested that various stimuli play a role in augmenting allergic reactions via modulation of activated basophil cytokine secretion. It may require new methods to stabilize the basophils in allergic diseases.

  18. Basophil sensitivity through CD63 or CD203c is a functional measure for specific immunotherapy

    Directory of Open Access Journals (Sweden)

    Dahl Ronald

    2010-02-01

    Full Text Available Abstract Background Subcutaneous Immunotherapy (SCIT modifies the allergic response and relieves allergic symptoms. SCIT is the only and a very effective treatment for insect venom allergy. We hypothesized that basophil sensitivity, measured through the basophil activation test, would decrease during SCIT up dosing. Expression of CD203c was compared to CD63 as marker for basophil activation, using a Bland Altman plot and ROC curves. Methods Patients (n = 18 starting subcutaneous SCIT for wasp allergy with an up dosing scheme of 7 to 11 weeks were enrolled. Heparinised blood samples were drawn at weeks 1-4, 7 and at the first maintenance visit. Basophils were stimulated at 7 log dilutions of V. vespula allergen for 15 min, and were stained with CD203c and CD63. Basophils were identified as CD203c+ leukocytes, and the proportion of CD63+ and CD203c+ cells were plotted against allergen concentration. A sigmoid curve was fitted to the points, and the allergen concentration at which half of the maximal activation was achieved, LC50, was calculated. In another series of experiments, LC50 calculated in whole blood (AP was subtracted from LC50 calculated with basophils suspended in plasma from a nonatopic donor (HS to determine the protective effect of soluble factors in blood of patients treated with SCIT. Results Heparin blood basophil activation was similar through CD63 and CD203c. Basophils were significantly more sensitized three weeks after initiation of SCIT compared to baseline (p Conclusion Basophil activation is a versatile and sensitive tool that measures changes in the humoral immune response to allergen during SCIT.

  19. Increased Level of Basophil CD203c Expression Predicts Severe Chronic Urticaria

    OpenAIRE

    Ye, Young-Min; Yang, Eun-Mi; Yoo, Hye-Soo; Shin, Yoo-Seob; Kim, Seung-Hyun; Park, Hae-Sim

    2013-01-01

    Increased FcεR1α expression with upregulated CD203c expression on peripheral basophils is seen in patients with chronic urticaria (CU). However, there has been no published report on the association between CD203c expression level and clinical disease activity in CU patients. To investigate whether the increase of basophil activation is associated with the disease activity of CU, we measured basophil CD203c expression using a tricolor flow cytometric method in 82 CU patients and 21 normal con...

  20. Difference in CD22 molecules in human B cells and basophils

    OpenAIRE

    Toba, Ken; Hanawa, Haruo; Fuse, Ichiro; Sakaue, Minori; Watanabe, Kenichi; Uesugi, Yumiko; Higuchi, Wataru; Takahashi, Wataru; Aizawa, Yoshifusa; 鳥羽, 健

    2002-01-01

    Objective. CD22 is believed to be restricted to normal and neoplastic B cells. Human basophils were found to express CD22 molecules. Among the antibodies against CD22, Leu14, which recognized the ligand binding domain reacted to basophils, and B3 and 4KB128, which recognized the amino terminus side and carboxy terminus side of the ligand binding epitope, respectively, did not. To clarify the difference of CD22 antigenicity in human B cells and basophils, we investigated RNA sequence and struc...

  1. Trisomy 19 and T(9;22 In a Patient with Acute Basophilic Leukemia

    Directory of Open Access Journals (Sweden)

    Alicia Rojas-Atencio

    2011-01-01

    Full Text Available We report a case of acute basophilic leukemia with two coexisting clonal abnormalities, t(9;22 and trisomy 19. The blast showed positive reaction with myeloperoxidase but negative reaction with chloroacetate esterase and acid phosphatase. Metachromatic features of the blast were observed with toluidine blue stain. Ultrastructure study showed the presence of azurophilic granules in basophils and blast mast cells. Conventional and molecular cytogenetic studies revealed, t(9;22 with BCR/ABL positive and trisomy 19 in all metaphase cells. To our knowledge, this paper here is the first to present acute basophilic leukemia with trisomy 19 and t(9;22.

  2. The Evolution of Human Basophil Biology from Neglect towards Understanding of Their Immune Functions

    Science.gov (United States)

    Steiner, Markus; Huber, Sara; Harrer, Andrea

    2016-01-01

    Being discovered long ago basophils have been neglected for more than a century. During the past decade evidence emerged that basophils share features of innate and adaptive immunity. Nowadays, basophils are best known for their striking effector role in the allergic reaction. They hence have been used for establishing new diagnostic tests and therapeutic approaches and for characterizing natural and recombinant allergens as well as hypoallergens, which display lower or diminished IgE-binding activity. However, it was a long way from discovery in 1879 until identification of their function in hypersensitivity reactions, including adverse drug reactions. Starting with a historical background, this review highlights the modern view on basophil biology. PMID:28078302

  3. Increasing time interval and decreasing allergen dose interval improves ex vivo desensitization of human blood basophils

    DEFF Research Database (Denmark)

    Witting Christensen, Sara K; Krohn, Inge Kortekaas; Thuraiaiyah, Jani;

    2016-01-01

    BACKGROUND: Desensitization is a method for inducing temporary tolerance to allergen. The mechanism underlying desensitization is yet to be established. METHODS: Basophil granulocytes in whole blood from grass pollen allergic subjects were desensitized ex vivo by sequential addition of increasing...... allergen concentrations. At each step basophil activation (CD193(+) CD63(+) ) was monitored with and without (background activation) allergen challenge at optimal concentration. The sequential desensitization protocol was compared to a single-dose desensitization protocols with threshold and subthreshold...... allergen concentrations. Incubation intervals and allergen concentrations were varied in order to optimise the protocol. RESULTS: Sequential desensitization effectively reduced basophil response. The single-dose subthreshold protocol and single-dose threshold protocols did not reduce basophil activation...

  4. Surface profiling of normally responding and nonreleasing basophils by flow cytometry

    DEFF Research Database (Denmark)

    Kistrup, Kasper; Poulsen, Lars Kærgaard; Jensen, Bettina Margrethe

    Background Human basophils are granulocytes with the capacity to play important roles in allergy for example by releasing histamine when activated by cross-linking of their high affinity IgE receptors (Fc¿RI). However, not all individuals have basophils responding with a histamine release after......c, C3aR, C5aR CCR3, FPR1, ST2, CRTH2 on anti-IgE respondsive and nonreleasing basophils by flow cytometry, thereby generating a surface profile of the two phenotypes. Methods Fresh buffy coat blood (histamine release and nonreleases were defined as having...... a maximum release basophils, defined as FceRIa+CD3-CD14-CD19-CD56-,were analysed for surface expression of relevant markers. All samples were compensated and analysed in logicle display. All gates...

  5. Aspirin Augments IgE-Mediated Histamine Release from Human Peripheral Basophils via Syk Kinase Activation

    Directory of Open Access Journals (Sweden)

    Hiroaki Matsuo

    2013-01-01

    Conclusions: Aspirin enhanced histamine release from basophils via increased Syk kinase activation, and that the augmentation of histamine release by NSAIDs or FAs may be one possible cause of worsening symptoms in patients with chronic urticaria and FDEIA.

  6. The basophil activation test by flow cytometry: recent developments in clinical studies, standardization and emerging perspectives

    OpenAIRE

    Debard Anne-Lise; Boumiza Radhia; Monneret Guillaume

    2005-01-01

    Abstract The diagnosis of immediate allergy is mainly based upon an evocative clinical history, positive skin tests (gold standard) and, if available, detection of specific IgE. In some complicated cases, functional in vitro tests are necessary. The general concept of those tests is to mimic in vitro the contact between allergens and circulating basophils. The first approach to basophil functional responses was the histamine release test but this has remained controversial due to insufficient...

  7. Responsiveness of basophil granulocytes of horses suffering from chronic obstructive pulmonary disease to various allergens.

    Science.gov (United States)

    Dirscherl, P; Grabner, A; Buschmann, H

    1993-10-01

    As basophils are the major effector cells of allergic reactions, confirmation of the allergic etiology of chronic obstructive pulmonary disease (COPD) was sought by the demonstration of a specific in vitro response of equine basophilic blood cells to some potential allergens (Aspergillus, Cladosporidium, Mucor, Penicillium, extracts of dust particles of hay and straw). The allergen induced degranulation of basophils and the histamine and protease release from basophils during incubation with the allergens were tested. By evaluating the results obtained from 14 COPD horses and eight controls it could be shown that the sensitivity of the basophils of affected horses was increased, particularly against the allergen extract of Mucor mucedo and Mucor spinosus. Further a greater percentage of COPD horses reacted positively with the Mucor allergen extract. The mitogenic stimulation of lymphocytes by PHA and by the allergen extracts used gave comparable results in affected and control horses. Thus the in vitro stimulation of basophils may be an easily to perform testing device for the identification of potential allergens involved in the pathogenesis of equine COPD.

  8. Mutual interaction of Basophils and T cells in chronic inflammatory diseases

    Directory of Open Access Journals (Sweden)

    Marika eSarfati

    2015-08-01

    Full Text Available Basophils are, together with mast cells, typical innate effector cells of allergen-induced IgE-dependent allergic diseases. Both cell types express the high affinity receptor for IgE (FcεR1, release histamine, inflammatory mediators and cytokines following FcεR1 cross-linking. Basophils are rare granulocytes in blood, lymphoid and non-lymphoid tissues and the difficulties to detect and isolate these cells has hampered the study of their biology and the understanding of their possible role in pathology. Furthermore, the existence of other FcεR1-expressing cells, including professional Ag-presenting dendritic cells, generated some controversy regarding the ability of basophils to express MHC Class II molecules, present Ag and drive naïve T cell differentiation into Th2 cells. The focus of this review is to present the recent advances on the interactions between basophils and peripheral blood and tissue memory Th1, Th2 and Th17 cells, as well as their potential role in IgE-independent non allergic chronic inflammatory disorders, including human inflammatory bowel diseases. Basophils interactions with the innate players of IgE-dependent allergic inflammation, particularly innate lymphoid cells, will also be considered. The previously unrecognized function for basophils in skewing adaptive immune responses opens novel perspectives for the understanding of their contribution to the pathogenesis of inflammatory diseases.

  9. IgE-mediated 15-hydroxyeicosatetraenoic acid (15-HETE) generation by peripheral blood leukocytes: its association with basophil activation

    OpenAIRE

    Michalak, Anna; Lewandowska-Polak, Anna; Moskwa, Sylwia; Kowalski, Marek L; Grzegorczyk, Janina Ł.

    2015-01-01

    Introduction Allergen-induced basophil activation has been associated with the release of several mediators and with an increased expression of CD203c molecules on basophils. Aim To assess the influence of specific allergens on the generation of 15-hydroxyeicosatetraenoic (15-HETE) from peripheral blood leukocytes in relation to basophil activation, on the basis of CD203c molecule expression and histamine release. Material and methods The study included 15 patients with clinical symptoms of b...

  10. Basophil count, a marker for disease activity in systemic lupus erythematosus.

    Science.gov (United States)

    Liang, Peifen; Tang, Ying; Fu, Sha; Lv, Jun; Liu, Bo; Feng, Min; Li, Jinggao; Lai, Deyuan; Wan, Xia; Xu, Anping

    2015-05-01

    Systemic lupus erythematosus (SLE) is a chronic multisystem autoimmune disease, with frequent flares amid remissions. Basophils contribute to the immunopathogenesis of SLE. This retrospective clinical study evaluated blood basophil count as a potential marker of SLE activity. This study included 213 patients with SLE, 70 with non-SLE chronic kidney disease (CKD), and 100 healthy volunteers. SLE disease activity was scored using the SLE Disease Activity Index (SLEDAI). Baseline and post-immunosuppressant bioparameters were compared in patients with active SLE, with second samples taken at total SLEDAI ≤4. Blood basophil counts and other conventional biomarkers were compared among the groups. Among the 213 SLE patients (192 women, 21 men; mean age 33.0 ± 12.0 years), 149 had active disease. Basophil counts were significantly lower in patients with SLE than in patients with non-SLE CKD and healthy controls (0.009 ± 0.010 vs. 0.025 ± 0.015 vs. 0.022 ± 0.010 × 10(9)/L, p Basophil counts in SLE patients were significantly higher after than before immunosuppressive treatment (0.021 ± 0.017 vs. 0.008 ± 0.008 × 10(9)/L, p analysis showed that basophil counts were similar to conventional markers (leukocytes, platelets, and double-stranded (ds) DNA IgG) in differentiating active from inactive SLE. These findings indicate that blood basophil counts may be a useful biomarker in evaluating SLE activity.

  11. Evaluation of the correlation and reproducibility between histamine, IL-4, and IL-13 release from human basophils.

    OpenAIRE

    Nahid Eskandari; Reza Bastan; Maryam Ahmadi; Peachell, Peter T

    2014-01-01

    Human basophils play a key role in allergic diseases such as asthma and in a variety of immunological disorders. The generation of IL-4 and IL-13 can be induced from basophil by IgE-mediated and non-IgE-mediated mechanisms. Time and stimulus-dependent differences in the regulation of these cytokines could have relevance to their biological effects. The aim of the present study was activation of basophils in order to evaluate the extent of histamine, IL-4, and IL-13 generations. Basophil-enric...

  12. Analysis of basophil activation by flow cytometry in pediatric house dust mite allergy.

    Science.gov (United States)

    González-Muñoz, Miguel; Villota, Julian; Moneo, Ignacio

    2008-06-01

    Detection of allergen-induced basophil activation by flow cytometry has been shown to be a useful tool for allergy diagnosis. The aim of this study was to assess the potential of this technique for the diagnosis of pediatric house dust mite allergy. Quantification of total and specific IgE and basophil activation test were performed to evaluate mite allergic (n = 24), atopic (n = 23), and non-allergic children (n = 9). Allergen-induced basophil activation was detected as a CD63-upregulation. Receiver operating characteristics (ROC) curve analysis was performed to calculate the optimal cut-off value of activated basophils discriminating mite allergic and non-allergic children. ROC curve analysis yielded a threshold value of 18% activated basophils when mite-sensitized and atopic children were studied [area under the curve (AUC) = 0.99, 95% confidence interval (CI) = 0.97-1.01, p Analysis of the data obtained with 1.6 microg/ml mite extract defined a cut-off value of 8% activated basophils (AUC = 0.96, 95% CI = 0.91-1.01; p basophils (AUC = 1.0) with 16 microg/ml allergen extract and a sensitivity and specificity of 100%. The same threshold and specificity values were obtained with 1.6 microg/ml extract (AUC = 97%, 95% CI = 0.92-1.02; p basophil activation tests and high specific IgE (>43 kU/l) values for Dermatophagoides pteronyssinus allergen. They also showed positive prick (wheal diameter >1.0 cm) and basophil activation (>87%) tests and high specific IgE (>100 kU/l) with shrimp allergen. Shrimp sensitization was demonstrated by high levels of Pen a 1-specific IgE (>100 kU/l). Cross-reactivity between mite and shrimp was confirmed by fluorescence enzyme immunoassay (FEIA-CAP) inhibition study in these two cases. This study demonstrated that the analysis of allergen-induced CD63 upregulation by flow cytometry is a reliable tool for diagnosis of mite allergy in pediatric patients, with sensitivity similar to routine diagnostic tests and a higher specificity

  13. Impact of thermal processing and the Maillard reaction on the basophil activation of hazelnut allergic patients.

    Science.gov (United States)

    Cucu, Tatiana; De Meulenaer, Bruno; Bridts, Chris; Devreese, Bart; Ebo, Didier

    2012-05-01

    Food allergy, an abnormal immunological response due to sensitization to a food component, has become an important health problem, especially in industrialized countries. The aim of this study was to investigate the impact of thermal processing and glycation on the basophil activation by hazelnut proteins using a basophil activation test. Patients with systemic allergic reactions (SR; n=6) to hazelnut as well as patients with an isolated oral allergy syndrome (OAS; n=4) were investigated. Thermal processing of hazelnut proteins either in the presence or absence of wheat proteins did not result in major changes in the stimulatory activity of the basophils for patients with SR or OAS. For the patients with OAS, incubation of hazelnut proteins with glucose led to complete depletion of the stimulatory activity of the basophils. An increase in stimulatory activity of the basophils for two out of six patients with SR was observed. For the other four patients slight or complete abolition of the stimulatory activity was observed. These results indicate that some patients with SR to hazelnut are at risk when exposed to hazelnut proteins, even in processed foods.

  14. Surface membrane traffic in guinea pig basophils exposed to cationic ferritin.

    Science.gov (United States)

    Dvorak, A M; Dvorak, H F; Galli, S J

    1985-01-01

    Surface membrane traffic patterns can be influenced by a number of factors, including the functional state of the cell. We used transmission electron microscopy to investigate the fate of surface membrane in guinea pig basophils exposed to cationized ferritin (CF) in vitro. CF bound to the plasma membrane and was internalized on the membranes of vesicles and vacuoles, a process that was particularly prominent at the uropod of basophils exhibiting a polarized ('motile') configuration. The vesicles/vacuoles moved to the Golgi area, or, in the case of degranulating basophils, were observed in continuity with the degranulation sac, a structure formed largely by the fusion of individual cytoplasmic granule membranes. However, CF-positive vesicles were never observed to fuse directly with the membranes of intact cytoplasmic granules.

  15. Expression of the Transcription Factor E4BP4 in Human Basophils

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Gohr, Maria; Poulsen, Lars Kærgaard

    2014-01-01

    by ECL stain and X-ray film exposure. Protein band intensities were correlated to -actin expression. Results We analyzed basophils from 14 donors and found E4BP4 mRNA expression in all donors (2.33 ± 2.42) despite a low basophil RNA level. Seven donors were also tested for E4BP4 protein expression...... by Alcian blue. RNA was extracted (0.005-0.02 µg RNA from 0.5 - 1 x 106 cells), and the corresponding cDNA analyzed by real-time PCR where E4BP4 expression was calculated as 2-(CT(E4BP4) - CT(β-actin)). E4BP4 protein expression was visualized in basophil lysates (107 cells/ml) by Western blot followed...

  16. Novel basophil- or eosinophil-depleted mouse models for functional analyses of allergic inflammation.

    Science.gov (United States)

    Matsuoka, Kunie; Shitara, Hiroshi; Taya, Choji; Kohno, Kenji; Kikkawa, Yoshiaki; Yonekawa, Hiromichi

    2013-01-01

    Basophils and eosinophils play important roles in various host defense mechanisms but also act as harmful effectors in allergic disorders. We generated novel basophil- and eosinophil-depletion mouse models by introducing the human diphtheria toxin (DT) receptor gene under the control of the mouse CD203c and the eosinophil peroxidase promoter, respectively, to study the critical roles of these cells in the immunological response. These mice exhibited selective depletion of the target cells upon DT administration. In the basophil-depletion model, DT administration attenuated a drop in body temperature in IgG-mediated systemic anaphylaxis in a dose-dependent manner and almost completely abolished the development of ear swelling in IgE-mediated chronic allergic inflammation (IgE-CAI), a typical skin swelling reaction with massive eosinophil infiltration. In contrast, in the eosinophil-depletion model, DT administration ameliorated the ear swelling in IgE-CAI whether DT was administered before, simultaneously, or after, antigen challenge, with significantly lower numbers of eosinophils infiltrating into the swelling site. These results confirm that basophils and eosinophils act as the initiator and the effector, respectively, in IgE-CAI. In addition, antibody array analysis suggested that eotaxin-2 is a principal chemokine that attracts proinflammatory cells, leading to chronic allergic inflammation. Thus, the two mouse models established in this study are potentially useful and powerful tools for studying the in vivo roles of basophils and eosinophils. The combination of basophil- and eosinophil-depletion mouse models provides a new approach to understanding the complicated mechanism of allergic inflammation in conditions such as atopic dermatitis and asthma.

  17. Novel basophil- or eosinophil-depleted mouse models for functional analyses of allergic inflammation.

    Directory of Open Access Journals (Sweden)

    Kunie Matsuoka

    Full Text Available Basophils and eosinophils play important roles in various host defense mechanisms but also act as harmful effectors in allergic disorders. We generated novel basophil- and eosinophil-depletion mouse models by introducing the human diphtheria toxin (DT receptor gene under the control of the mouse CD203c and the eosinophil peroxidase promoter, respectively, to study the critical roles of these cells in the immunological response. These mice exhibited selective depletion of the target cells upon DT administration. In the basophil-depletion model, DT administration attenuated a drop in body temperature in IgG-mediated systemic anaphylaxis in a dose-dependent manner and almost completely abolished the development of ear swelling in IgE-mediated chronic allergic inflammation (IgE-CAI, a typical skin swelling reaction with massive eosinophil infiltration. In contrast, in the eosinophil-depletion model, DT administration ameliorated the ear swelling in IgE-CAI whether DT was administered before, simultaneously, or after, antigen challenge, with significantly lower numbers of eosinophils infiltrating into the swelling site. These results confirm that basophils and eosinophils act as the initiator and the effector, respectively, in IgE-CAI. In addition, antibody array analysis suggested that eotaxin-2 is a principal chemokine that attracts proinflammatory cells, leading to chronic allergic inflammation. Thus, the two mouse models established in this study are potentially useful and powerful tools for studying the in vivo roles of basophils and eosinophils. The combination of basophil- and eosinophil-depletion mouse models provides a new approach to understanding the complicated mechanism of allergic inflammation in conditions such as atopic dermatitis and asthma.

  18. A comparative study of the FcepsilonRI molecule on human mast cell and basophil cell lines

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Dissing, S; Skov, P S;

    2005-01-01

    Mast cells and basophils express the high-affinity IgE receptor FcepsilonRI. We have analysed the human mast cell line LAD2 and four subclones of the basophil cell line KU812 in order to reveal possible differences concerning the FcepsilonRI surface regulation, anti-IgE-triggered activation...

  19. L3L4ES antigen and secretagogues induce histamine release from porcine peripheral blood basophils after Ascaris suum infection

    Science.gov (United States)

    The aim of this paper was to investigate the role of porcine basophils in protective immunity. Experimental pigs were infected with 1,000 Ascaris suum eggs daily for 21 days. Control pigs were maintained helminth-free. Circulating porcine basophils were isolated from the anti-coagulated whole blood ...

  20. Human blood basophils display a unique phenotype including activation linked membrane structures.

    Science.gov (United States)

    Stain, C; Stockinger, H; Scharf, M; Jäger, U; Gössinger, H; Lechner, K; Bettelheim, P

    1987-12-01

    To evaluate the membrane marker profile of human basophils a panel of well-established monoclonal antibodies (MoAbs, n = 60) was used for a combined toluidine/immunofluorescence staining procedure. Myeloid-associated MoAbs (particularly MoAbs against the LFA-1 family (CD11, CDw18), MoAbs directed against lactosylceramide (CDw17), anti-glycoprotein (gp) 150 MoAbs MCS 2 and MY 7 (CDw13), anti-gp 67 MoAb MY 9, anti Fc gamma-receptor (mol wt 40 kd) MoAb CIKM5, anti-CR 1 MoAb E 11, and the antiglycolipid MoAb VIM-2) were reactive with basophils, indicating a close relationship to other mature myeloid cells. Under normal conditions, basophils surprisingly express at least three activation-linked structures not detectable on mature neutrophils, ie, the p45 structure defined by MoAbs OKT-10 and VIP-2b, the p24 structure identified by the CD9 MoAb BA-2, and the receptor for interleukin 2 (IL 2) recognized by three different MoAbs (anti-TAC, IL2RI, anti-IL 2). Moreover, under short-term culture conditions basophils both in mononuclear cell (MNC) suspension and as purified fractions display the HLA-DR and T4 antigens. The neutrophilic/eosinophilic structure 3-fucosyl-N-acetyllactosamine is expressed on basophils only after neuraminidase treatment. Basophils were not stained at all by CD 16 MoAbs directed against the Fc gamma-receptor (mol wt 50 to 70 kd) of neutrophils, by the MoAb 63D3 (CDw12) recognizing the monocyte/granulocyte-associated p 200 antigen, and by the CDw 14 antibodies (VIM-13, Mo 2) defining the monocyte-specific structure p 55. Enriched basophils freshly obtained from chronic granulocytic leukemia (CGL) patients yielded identical results in FACS analyses. In summary, these data indicate that basophils generate a unique combination of surface determinants and possibly represent an activated cell population.

  1. Mouse Basophils Reside in Extracellular Matrix-Enriched Bone Marrow Niches Which Control Their Motility

    OpenAIRE

    Salete Smaniotto; Elke Schneider; Nicolas Goudin; Rachel Bricard-Rignault; François Machavoine; Mireille Dardenne; Michel Dy; Wilson Savino

    2013-01-01

    Basophils co-express FcεRIα and CD49b, the α-2 chain of integrin-type receptor VLA-2 (α2β1), which recognizes type-1 collagen as a major natural ligand. The physiological relevance of this integrin for interactions with extracellular bone marrow matrix remains unknown. Herein, we examined the expression of several receptors of this family by bone marrow-derived basophils sorted either ex-vivo or after culture with IL-3. Having established that both populations display CD49d, CD49e and CD49f (...

  2. Membrane sialic acid influences basophil histamine release by interfering with calcium dependence

    DEFF Research Database (Denmark)

    Jensen, C; Norn, S; Skov, P S

    1987-01-01

    The influence of the cell membrane content of sialic acid on basophil histamine release was examined in vitro in allergic patients and normal controls. Enzymatical removal of sialic acid enhanced histamine release induced by allergen and anti-IgE, whereas an increase in membrane sialic acid content....... This difference, together with the previous finding that alterations in membrane sialic acid content is reflected in the cell sensitivity to extracellular calcium, suggest an interaction between membrane sialic acid and the calcium channels involved in basophil histamine release....

  3. IN SUBJECTS ALLERGIC TO GRASS POLLEN, BASOPHIL SENSITIVITY DECREASES DURING SUBCUTANEOUS IMMUNOTHERAPY DUE TO BOTH HUMORAL FACTORS AND CELLULAR DESENSITIZATION

    DEFF Research Database (Denmark)

    Schmid, Johannes Martin; Dahl, Ronald; Hoffmann, Hans Jürgen

    2011-01-01

    with allergic rhino-conjunctivitis with basophil activation tests (BAT). Methods: We have randomized 24 patients to a treatment (18) and an open control group (6). Repeated BAT were performed at baseline and througout treatment. Heparinized blood was centrifuged, plasma was removed, the cells washed twice......, then reconstituted with plasma from the present and baseline visit, and medium. The cells were 406 ESCCA AND ICyS ABSTRACTS Cytometry Part B: Clinical Cytometry incubated with log dilutions of grass pollen extract and washed again. Basophil activation was measured as the percentage of CD63 expression on CD193 þ...... basophils. The LC50 was calculated as the log of the allergen concentration leading to half maximum basophil activation. Specific IgE and IgG4 was measured using ISAC panel. Results: We found a 15-fold reduction in basophil sensitivity in the treated group during updosing, compared to their baseline...

  4. Some invariant properties of IgE-mediated basophil activation and desensitization

    Energy Technology Data Exchange (ETDEWEB)

    Goldstein, B.; Dembo, M.; Sobotka, A.K.; Lichtenstein, L.M.

    1979-10-01

    We investigate certain general properties of antigen induced degranulation of sensitized basophils by analyzing two types of experiments: experiments in which we expose basophils to two antigens seuquentially and then determine the fraction of histamine released; and experiments in which we obtain time-dependent release and desensitization curves. To analyze the latter type of experiments we introduce a new way to plot release and desensitization data that depends on the nature of the interactions of histamine-containing units (histamine quanta) with themselves or the cells degranulation apparatus, but not on any specific properties of the antigen. From our analysis we conclude that: (1) a fraction of histamine within a population of basophils is nonreleasable by antigenic stimulation. (2) When a basophil degranulates the initial release of histamine appears to inhibit subsequent release. (3) The rate of histamine release is proportional to the amount of releasable histamine remaining in the cells when the amount remaining is small, as expected if release of histamine granules is a stochastic process. (4) There is no dependence of desensitization on the extracellular calcium concentration.

  5. Biochemical and functional characterization of proteoglycans isolated from basophils of patients with chronic myelogenous leukemia.

    Science.gov (United States)

    Metcalfe, D D; Bland, C E; Wasserman, S I

    1984-04-01

    Human basophils were obtained from three donors with myelogenous leukemia. Proteoglycans were labeled by using [35S]sulfate as precursor and were extracted in 1 M NaCl with protease inhibitors to preserve their native structure. [35S]proteoglycans filtered on Sepharose 4B with an average m.w. similar to that of a rat heparin proteoglycan that has an estimated m.w. of 750,000. The [35S]glycosaminoglycan side chains filtered with an average m.w. slightly smaller than a 60,000-m.w. glycosaminoglycan marker. The [35S]glycosaminoglycans were resistant to heparinase and susceptible to degradation by chondroitin AC lyase and chondroitin ABC lyase. The intact [35S]glycosaminoglycans chromatographed on DEAE Sepharose as a single peak eluting just before an internal heparin marker. These findings indicate that the [35S]glycosaminoglycans were made up only of chondroitin sulfates. No heparin was identified. The chondroitin sulfate disaccharides that resulted from the action of chondroitin ABC lyase on the basophil glycosaminoglycans consisted of 92% delta Di-4S, 6% delta Di-6S, and 2% disulfated disaccharides. The [35S]chondroitin sulfate proteoglycans were susceptible to cleavage with proteases and could be shown to be released intact from basophils during degranulation initiated by the calcium ionophore A23187. The basophil proteoglycans and glycosaminoglycans were capable of binding histamine in water, but not in phosphate-buffered saline, and had no anticoagulant activity.

  6. Ultrastructural localization of Charcot-Leyden crystal protein in human eosinophils and basophils.

    Science.gov (United States)

    Calafat, J; Janssen, H; Knol, E F; Weller, P F; Egesten, A

    1997-01-01

    The Charcot-Leyden crystal (CLC) protein with lysophospholipase activity and carbohydrate-binding properties is a characteristic constituent of eosinophils and basophils. We investigated its subcellular distribution using immunoelectron microscopy. Eosinophil progenitors, mature eosinophils and basophils all contained CLC protein in their cytosol and in the euchromatin of the nucleus. A minor population of granules in eosinophils, increasing in number with maturation, and a more abundant granule-population in basophils, were found to contain CLC protein. Double-labeling experiments showed, in eosinophils, that CLC protein-containing granules contain also eosinophil peroxidase, a characteristic specific granule protein. This suggests a relationship between the CLC protein-containing organelle and the specific granule. In basophils both the CLC protein positive and the negative granules showed the same characteristic particulate-like structure of the granular matrix and both share the same membrane marker CD63. In nasal polyps, macrophages were observed phagocytosing necrotic eosinophils. In these macrophages CLC protein-containing vesicles were observed, probably representing late endosomes. The dual (cytosolic/nuclear and granular) localization of CLC protein suggests that this protein enters both a secretory and a nonsecretory pathway during its biosynthesis, indicating functional roles for this protein both within the cell and extracellularly.

  7. Chemokine stromal cell-derived factor 1alpha activates basophils by means of CXCR4

    DEFF Research Database (Denmark)

    Jinquan, T; Jacobi, H H; Jing, C

    2000-01-01

    The CXC chemokine receptor 4 (CXCR4) is predominantly expressed on inactivated naive T lymphocytes, B lymphocytes, dendritic cells, and endothelial cells. CXC chemokine stromal cell-derived factor 1alpha (SDF-1alpha) is the only known ligand for CXCR4. To date, the CXCR4 expression and function...... of SDF-1alpha in basophils are unknown....

  8. A Case of Anaphylactic Reaction Following Matsutake Mushroom Ingestion: Demonstration of Histamine Release Reaction of Basophils

    Directory of Open Access Journals (Sweden)

    Takako Toda

    2010-01-01

    Discussion: Matsutake mushroom can, on rare occasions, cause anaphylaxis in sensitized people, a reaction so far observed only in Japan. Not ony the in vivo prick-to-prick test but also the in vitro basophil activation test utilizing the patient's blood represent useful methods for allergen identification and also for identification of sensitized subjects.

  9. Allergy to formaldehyde: basophil histamine-release test is useful for diagnosis.

    Science.gov (United States)

    Tanaka, Yusuke; Nakase, Yuko; Yamaguchi, Masao; Sugimoto, Naoya; Ohara, Kenshin; Nagase, Hiroyuki; Ohta, Ken

    2014-01-01

    We describe a case of formaldehyde-induced urticaria with a positive test result for serum IgE antibody against this substance. Formaldehyde's slow protein-binding property may explain why basophil histamine-release tests using fresh formaldehyde solutions are not diagnostic, whereas the tests are useful if formaldehyde that had been stored with albumin is used.

  10. Bimodal action of the flavonoid quercetin on basophil function: an investigation of the putative biochemical targets

    Directory of Open Access Journals (Sweden)

    Chirumbolo Salvatore

    2010-09-01

    Full Text Available Abstract Background Flavonoids, a large group of polyphenolic metabolites derived from plants have received a great deal of attention over the last several decades for their properties in inflammation and allergy. Quercetin, the most abundant of plant flavonoids, exerts a modulatory action at nanomolar concentrations on human basophils. As this mechanism needs to be elucidated, in this study we focused the possible signal transduction pathways which may be affected by this compound. Methods: K2-EDTA derived leukocyte buffy coats enriched in basophil granulocytes were treated with different concentrations of quercetin and triggered with anti-IgE, fMLP, the calcium ionophore A23187 and the phorbol ester PMA in different experimental conditions. Basophils were captured in a flow cytometry analysis as CD123bright/HLADRnon expressing cells and fluorescence values of the activation markers CD63-FITC or CD203c-PE were used to produce dose response curves. The same population was assayed for histamine release. Results Quercetin inhibited the expression of CD63 and CD203c and the histamine release in basophils activated with anti-IgE or with the ionophore: the IC50 in the anti-IgE model was higher than in the ionophore model and the effects were more pronounced for CD63 than for CD203c. Nanomolar concentrations of quercetin were able to prime both markers expression and histamine release in the fMLP activation model while no effect of quercetin was observed when basophils were activated with PMA. The specific phosphoinositide-3 kinase (PI3K inhibitor wortmannin exhibited the same behavior of quercetin in anti-IgE and fMLP activation, thus suggesting a role for PI3K involvement in the priming mechanism. Conclusions These results rule out a possible role of protein kinase C in the complex response of basophil to quercetin, while indirectly suggest PI3K as the major intracellular target of this compound also in human basophils.

  11. The Effect of Stimuli on Basophil-Mediated Atopic Responses During Asthmatic Lying-In Women and in Newborns

    OpenAIRE

    Yang, Ling; Guo, Yin-Shi; Jiang, Jin-Qi; Guo, Xue-jun; Yi-ping XU; Tian, Ye; Xiong, Ying; Han, Li

    2012-01-01

    Morbidity from allergic diseases is increasing. Basophils play a critical role in systemic anaphylaxis and chronic allergic inflammation. The prenatal environment must be regarded as a possible early risk factor for allergic diseases in children. Our objective was to determine if basophils harvested from neonates genetically predisposed to atopic disease had different levels of CD63 expression and IL-4 release properties in response to various stimuli (peptidoglycan, Dermatophagoides farinae,...

  12. Chronic myeloid leukemia presenting with absence of basophils and marked dyspoiesis

    Directory of Open Access Journals (Sweden)

    Anand M

    2003-01-01

    Full Text Available A 61-year old woman presented to us with fever, weakness and ecchymotic patches for one year. She had leucocytosis, anemia and thrombocytopenia. Peripheral blood smear showed 62% neutrophils, 32% myelocytes and metamyelocytes, 2% promyelocytes, 1% blasts, 2% monocytes, 1% lymphocytes but no basophils and marked dyspoiesis. Bone marrow picture was essentially the same. A diagnosis of atypical chronic myeloid leukemia was suggested. The correct diagnosis of chronic myeloid leukemia - accelerated phase was, however, made on cytogenetic analysis which showed Philadelphia chromosome (Ph and isochromosome 17q [i(17q]. This case describes a rare and diagnostically difficult presentation of CML arising out of a combination of prominent dyspoiesis and near absence of peripheral blood basophils.

  13. Basophil activation test by flow cytometry: present and future applications in allergology.

    Science.gov (United States)

    Ebo, D G; Bridts, C H; Hagendorens, M M; Aerts, N E; De Clerck, L S; Stevens, W J

    2008-07-01

    The diagnosis of allergic reactions in clinical practice rests upon both clinical history and the demonstration of specific immunoglobulin E (sIgE), either in the serum or via skin tests. However, for various reasons, identification of the offending allergen(s) is not always possible. Moreover, not all allergies are IgE-mediated. In an attempt to find reliable methods to investigate hypersensitivity reactions, histamine and sulfidoleukotriene release tests have long been introduced. However, relatively few comprehensive quality reports have been published so far. Upon challenge with a specific allergen, basophils not only secrete quantifiable bioactive mediators but also upregulate the expression of different markers which can be detected efficiently by flow cytometry using specific monoclonal antibodies. This review addresses the principals, particular technical aspects and pitfalls as well as the clinical and research applications of flow-assisted analysis of in vitro activated basophils.

  14. Regulation of the Il4 gene is independently controlled by proximal and distal 3' enhancers in mast cells and basophils.

    Science.gov (United States)

    Yagi, Ryouji; Tanaka, Shinya; Motomura, Yasutaka; Kubo, Masato

    2007-12-01

    Mast cells and basophils are known to be a critical interleukin 4 (IL-4) source for establishing Th2 protective responses to parasitic infections. Chromatin structure and histone modification patterns in the Il13/Il4 locus of mast cells were similar to those of IL-4-producing type 2 helper T cells. However, using a transgenic approach, we found that Il4 gene expression was distinctly regulated by individual cis regulatory elements in cell types of different lineages. The distal 3' element contained conserved noncoding sequence 2 (CNS-2), which was a common enhancer for memory phenotype T cells, NKT cells, mast cells, and basophils. Targeted deletion of CNS-2 compromised production of IL-4 and several Th2 cytokines in connective-tissue-type and immature-type mast cells but not in basophils. Interestingly, the proximal 3' element containing DNase I-hypersensitive site 4 (HS4), which controls Il4 gene silencing in T-lineage cells, exhibited selective enhancer activity in basophils. These results indicate that CNS-2 is an essential enhancer for Il4 gene transcription in mast cell but not in basophils. The transcription of the Il4 gene in mast cells and basophils is independently regulated by CNS-2 and HS4 elements that may be critical for lineage-specific Il4 gene regulation in these cell types.

  15. Complexity of the influence of gangliosides on histamine release from human basophils and rat mast cells

    DEFF Research Database (Denmark)

    Jensen, C; Svendsen, U G; Thastrup, Ole;

    1987-01-01

    The influence of exogenous addition of gangliosides on histamine release from human basophils and rat mast cells was examined in vitro. Gangliosides dose-dependently inhibited histamine release, and this inhibition was dependent on the ganglioside sialic acid content, since GT1b, having 3 sialic...... was reflected in the sensitivity of the cells to extracellular calcium, since inhibition of the release could be counteracted by increasing the extracellular concentration of calcium....

  16. Basophil activation test in the study of food and drug hypersensitivity reactions

    OpenAIRE

    Carrapatoso, I; Cadinha, S; Sanz, ML

    2005-01-01

    The increase in the prevalence of adverse reactions to foods and drugs represents a constant challenge to the development of new methods of diagnosis. A meta-analysis on published studies concerning the clinical usefulness of the Basophil activation test (BAT) in these reactions was performed. High sensibilities and specificities can be achieved if certain technical requirements are observed. BAT results have a positive and high significant correlations with other routine diagnostic ...

  17. Infiltration of hypertrophic esophageal smooth muscle by mast cells and basophils.

    Science.gov (United States)

    Tung, H N; Schulze-Delrieu, K; Shirazi, S

    1993-01-01

    Partial obstruction leads to chronic distension and muscular hypertrophy of the opossum esophagus. The smooth muscle cells of the circular muscle layer enlarge, become pleomorphic and are surrounded by an amorphous ground substance in the extracellular space. Here we describe the histological and ultrastructural features of a peculiar cellular infiltrate in the hypertrophic smooth muscle. The infiltrate consisted uniquely of mast cells and basophils. In per unit area, the number of mast cells increased from 0.9 +/- 0.1 cells in controls to 3.7 +/- 0.2 in hypertrophic smooth muscle; the corresponding numbers for basophils were 2.5 +/- 0.2 and 7.2 +/- 0.3 cells. Cells were seen primarily in the septal spaces of the circular muscle layer and at the interface of the circular and longitudinal muscle layer. The cytoplasm of basophils is normally packed with round and oval granules. The granules stain metachromatically and with varying intensity on Wright-Giemsa stains. On transmission electronmicroscopy, granules display a membrane and a great diversity in the structure of their luminal contents. In hypertrophic muscle, most granules were discharging their contents into the cytoplasm or extracellular space. The membranes of adjacent empty granules then fused to form a chain of vacuoles. Similar changes occurred also in the mast cells which differed from the basophil by their lack of nuclear lobulation and by the greater homogeneity of their cytoplasmic granules. It is possible that these inflammatory cells are involved in the reconstruction of the smooth muscle and its connective tissue which occur during esophageal distension and hypertrophy.

  18. Inhibition by adenosine of histamine and leukotriene release from human basophils.

    Science.gov (United States)

    Peachell, P T; Lichtenstein, L M; Schleimer, R P

    1989-06-01

    Adenosine inhibited the release of histamine and leukotriene C4 (LTC4) from immunologically-activated basophils in a dose-dependent manner. Structural congeners of adenosine also attenuated the elaboration of these two mediators from stimulated basophils and a rank order of potency for the inhibition was observed following the sequence 2-chloroadenosine greater than or equal to N-ethylcarboxamidoadenosine (NECA) greater than adenosine greater than or equal to R-phenylisopropyladenosine (R-PIA) greater than or equal to S-PIA. These same nucleosides modulated the generation of LTC4 more potently than the release of histamine. A number of methylxanthines, which are antagonists of cell surface adenosine receptors, reversed the inhibition by adenosine and its congeners of the release of both histamine and LTC4 to varying extents. Dipyridamole and nitrobenzylthioinosine (NBTI), agents that block the intracellular uptake of adenosine, antagonized the inhibition of histamine release by adenosine (and 2-chloroadenosine) but failed to reverse the attenuation of LTC4 generation by the nucleoside. These same uptake blockers were unable to antagonize the inhibitory effects of NECA on either histamine or LTC4 release. In purified basophils, NECA and R-PIA, and in that order of decreasing reactivity, increased total cell cyclic adenosine monophosphate (cAMP) levels and inhibited the stimulated release of mediators. In total, these results suggest that the basophil possesses a cell surface adenosine receptor which, on the basis of both pharmacological and biochemical criteria, most closely conforms to an A2/Ra-like receptor. However, in addition to an interaction at the cell surface, studies with agents that block the intracellular uptake of adenosine suggest that the nucleoside may also exert intracellular effects when countering the release of histamine (but not LTC4).

  19. High-Cholesterol Diet Disrupts the Levels of Hormones Derived from Anterior Pituitary Basophilic Cells.

    Science.gov (United States)

    Yang, J; Zhang, X; Liu, Z; Yuan, Z; Song, Y; Shao, S; Zhou, X; Yan, H; Guan, Q; Gao, L; Zhang, H; Zhao, J

    2016-03-01

    Emerging evidence shows that elevated cholesterol levels are detrimental to health. However, it is unclear whether there is an association between cholesterol and the pituitary. We investigated the effects of a high-cholesterol diet on pituitary hormones using in vivo animal studies and an epidemiological study. In the animal experiments, rats were fed a high-cholesterol or control diet for 28 weeks. In rats fed the high-cholesterol diet, serum levels of thyroid-stimulating hormone (TSH; also known as thyrotrophin), luteinising hormone (LH) and follicle-stimulating hormone (FSH) produced by the basophilic cells of the anterior pituitary were elevated in a time-dependent manner. Among these hormones, TSH was the first to undergo a significant change, whereas adrenocorticotrophic hormone (ACTH), another hormone produced by basophilic cells, was not changed significantly. As the duration of cholesterol feeding increased, cholesterol deposition increased gradually in the pituitary. Histologically, basophilic cells, and especially thyrotrophs and gonadotrophs, showed an obvious increase in cell area, as well as a potential increase in their proportion of total pituitary cells. Expression of the β-subunit of TSH, FSH and LH, which controls hormone specificity and activity, exhibited a corresponding increase. In the epidemiological study, we found a similar elevation of serum TSH, LH and FSH and a decrease in ACTH in patients with hypercholesterolaemia. Significant positive correlations existed between serum total cholesterol and TSH, FSH or LH, even after adjusting for confounding factors. Taken together, the results of the present study suggest that the high-cholesterol diet affected the levels of hormones derived from anterior pituitary basophilic cells. This phenomenon might contribute to the pituitary functional disturbances described in hypercholesterolaemia.

  20. Acute myeloid leukemia with basophilic differentiation in a 3-year-old Standardbred gelding

    Science.gov (United States)

    Furness, Mary Catherine; Setlakwe, Emile; Sallaway, John; Wood, Darren; Fromstein, Jordan; Arroyo, Luis G.

    2016-01-01

    A 3-year-old Standardbred gelding with a history of pyrexia, persistent hemorrhage from the oral cavity, and a large, soft swelling at the junction of the caudal aspect of the mandibular rami and proximal neck was evaluated. The horse had neutropenia and anemia, with atypical granulated cells in a blood smear. Additional tests confirmed acute myeloid leukemia with basophilic differentiation, which has been reported in humans, cats, dogs, and cattle but not horses. PMID:27708445

  1. A Case of Anaphylactic Reaction Following Matsutake Mushroom Ingestion: Demonstration of Histamine Release Reaction of Basophils

    OpenAIRE

    Takako Toda; Masao Yamaguchi; Yuko Nakase; Naoya Sugimoto; Maho Suzukawa; Hiroyuki Nagase; Ken Ohta

    2010-01-01

    Background: Matsutake mushroom is not recognized as a common food allergen. However, several case reports have suggested that this mushroom can induce anaphylaxis on rare occasions. Case Summary: We report a woman with bronchial asthma, who experienced two episodes of Matsutake-induced anaphylaxis. Both the prick-to-prick test and basophil histamine release test showed positive reactions to this mushroom in this patient, but not in control subjects. Discussion: Matsutake mushroom can, o...

  2. Protein L. A bacterial Ig-binding protein that activates human basophils and mast cells.

    Science.gov (United States)

    Patella, V; Casolaro, V; Björck, L; Marone, G

    1990-11-01

    Peptostreptococcus magnus strain 312 (10(6) to 10(8)/ml), which synthesizes a protein capable of binding to kappa L chains of human Ig (protein L), stimulated the release of histamine from human basophils in vitro. P. magnus strain 644, which does not synthesize protein L, did not induce histamine secretion. Soluble protein L (3 x 10(-2) to 3 micrograms/ml) induced histamine release from human basophils. The characteristics of the release reaction were similar to those of rabbit IgG anti-Fc fragment of human IgE (anti-IgE): it was Ca2(+)- and temperature-dependent, optimal release occurring at 37 degrees C in the presence of 1.0 mM extracellular Ca2+. There was an excellent correlation (r = 0.82; p less than 0.001) between the maximal percent histamine release induced by protein L and that induced by anti-IgE, as well as between protein L and protein A from Staphylococcus aureus (r = 0.52; p less than 0.01). Preincubation of basophils with either protein L or anti-IgE resulted in complete cross-desensitization to a subsequent challenge with the heterologous stimulus. IgE purified from myeloma patients PS and PP (lambda-chains) blocked anti-IgE-induced histamine release but failed to block the histamine releasing activity of protein L. In contrast, IgE purified from myeloma patient ADZ (kappa-chains) blocked both anti-IgE- and protein L-induced releases, whereas human polyclonal IgG selectively blocked protein L-induced secretion. Protein L acted as a complete secretagogue, i.e., it activated basophils to release sulfidopeptide leukotriene C4 as well as histamine. Protein L (10(-1) to 3 micrograms/ml) also induced the release of preformed (histamine) and de novo synthesized mediators (leukotriene C4 and/or PGD2) from mast cells isolated from lung parenchyma and skin tissues. Intradermal injections of protein L (0.01 to 10 micrograms/ml) in nonallergic subjects caused a dose-dependent wheal-and-flare reaction. Protein L activates human basophils and mast cells in

  3. 578 Comparison of the Modified Autologous Serum Skin Test and the Cd63 Basophil Activation Test in Chronic Urticaria

    OpenAIRE

    Irinyi, B; Gyimesi, E.; Garaczi, E.; Bata, Zs.; Hodosi, K.; Zeher, M; Remenyik, É.; Szegedi, A.

    2012-01-01

    Background The modified CD63 basophil activation test in the diagnosis of chronic autoimmun urticaria was first described in 2004 by Szegedi et al. We demonstrated that the strongly sensitized basophils of atopic donors can be successfully used without the addition of IL-3 for the in vitro evaluation of autoimmun urticaria. Positive correlation was found between the basophil CD63 expression test and the autolog serum skin test (ASST), and between the CD63 test and the gold standard histamine ...

  4. Influence of nimodipine, verapamil and lanthanum on histamine release from human basophils

    DEFF Research Database (Denmark)

    Jensen, C B; Thastrup, Ole; Norn, S;

    1987-01-01

    Our previous studies suggest that the membrane content of sialic acid influences histamine release from human basophils by interfering with the transmembraneous calcium fluxes preceding histamine release. In this study we investigated a possible interaction between membrane sialic acid...... and the calcium channels, using the calcium antagonists nimodipine, verapamil and lanthanum. Anti-IgE-induced histamine release was inhibited by verapamil, nimodipine and lanthanum. When cells were pretreated with sialidase in order to remove sialic acid from the cell membrane, the inhibitory action of nimodipine...

  5. Effect of allergen-specific immunotherapy on recombinant human interleukin 3-mediated amplification of allergen-induced basophil histamine release.

    Science.gov (United States)

    Kowal, Krzysztof; Nolte, Hendrik; Skov, Per Stahl; DuBuske, Lawrence M

    2005-01-01

    Decreased allergen-induced histamine release from peripheral blood basophils in allergic rhinitis patients treated with specific immunotherapy (SIT) correlates with clinical outcomes of SIT. The aim of this study was to investigate if decreased histamine release is a permanent effect of SIT. Fifty-one patients (mean age, 35.3 years) with allergic rhinitis, diagnosed based on clinical history and positive skin-prick test results to common aeroallergens, were studied. Twenty-three patients had never received SIT (group A), and 28 patients had been treated with inhalant allergen extracts (group B). Eleven patients from group A participated in a prospective part of this study. Basophil histamine release in these patients was evaluated before (TO) and after-1 year (TI) of SIT. Histamine release from peripheral blood with and without interleukin (IL)-3 pretreatment was performed using the glass-fiber-based histamine release test. Brief pretreatment of whole blood basophils with one of the four concentrations (0.01, 0.1, 1, or 10 ng/mL) of recombinant human IL(rhIL)-3, rhIL-5, or rh-granulocyte-macrophage colony-stimulating factor resulted in a significant amplification of allergen-induced basophil histamine release. The amplification using cytokines at the optimal concentrations was the greatest with rhIL-3 and the lowest with rhIL-5; therefore, for further studies rhIL-3 was used. Prospective analysis showed no significant difference in allergen-induced basophil histamine release on rhIL-3 pretreatment after 1 year of SIT (192.7 +/- 75.3 ng and 176.1 +/- 76.4 ng for T0 and T1, respectively; p = 0.18). Short-term SIT does not decrease rhIL-3-mediated amplification of allergen-induced histamine release from peripheral blood basophils.

  6. Monomeric immunoglobulin E stabilizes FcepsilonRIalpha from the human basophil cell line KU812 by protecting it from natural turnover

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Hansen, Jens Bo; Dissing, S;

    2003-01-01

    The high affinity IgE receptor (FcepsilonRI) on mast cells and basophils is up-regulated by its own ligand IgE; however, the mechanism is unknown.......The high affinity IgE receptor (FcepsilonRI) on mast cells and basophils is up-regulated by its own ligand IgE; however, the mechanism is unknown....

  7. Role of protein kinase C in histamine release from human basophils.

    Science.gov (United States)

    Morita, Y; Takaishi, T; Honda, Z; Miyamoto, T

    1988-02-01

    In this study, we investigated the role of calcium and phospholipid-dependent protein kinase (protein kinase C, PKC) in the modulation of histamine release from human basophils. A novel and potent inhibitor of PKC, K-252a, inhibited the release of histamine induced by anti-IgE in a dose-dependent manner with ID50 (the dose required for 50% inhibition of histamine release) of 2.2 x 10(-8) M. Histamine release stimulated with 12-0-tetradecanoyl-phorbol-13-acetate(TPA) was also suppressed by K-252a with maximal inhibition of 48.0 +/- 9.3% at 10(-7) M. In contrast, K-252a did not inhibit the release of histamine in response to FMLP and ionophore A23187. Another inhibitor of PKC, H-7, exhibited a dose-dependent inhibition of anti-IgE-induced histamine release with ID50 of 8.6 x 10(-4) M. H-8 and HA1004, which closely resemble H-7 in chemical structure but are less potent in inhibiting PKC, did not inhibit histamine release stimulated with anti-IgE, but rather enhanced the release at higher concentrations. These results strongly suggest that PKC activation plays a crucial role in the mediation of IgE-mediated histamine release from human basophils.

  8. Diagnostic tests based on human basophils: more potentials and perspectives than pitfalls. II. Technical issues.

    Science.gov (United States)

    De Week, A L; Sanz, M L; Gamboa, P M; Aberer, W; Bienvenu, J; Blanca, M; Demoly, P; Ebo, D G; Mayorga, L; Monneret, G; Sainte Laudy, J

    2008-01-01

    Cellular basophil activation tests (BAT) such as histamine or sulfidoleukotriene-release tests for allergy diagnosis have been available for some time, but expression of basophil-activation markers such as CD63 and CD203c detected by flow cytometry has attracted particular attention in recent years. Not only the potential but also the possible pitfalls of flow-cytometric BAT have been stressed recently. Some authors have suggested that the technical problems are still such that BAT should only be performed in specialist laboratories. In an earlier review based on our clinical experience obtained over several years, we showed that, even using different protocols, reproducible and meaningful clinical results can be obtained. In this paper, we review the current knowledge in relation to several technical issues and show that flow-cytometric BAT already represents a major advance in the field of in vitro allergy diagnosis. We conclude that there are no serious technical justifications for depriving allergic patients of clinically indicated BAT tests, which can be performed reliably by any laboratory with the appropriate experience in allergy diagnosis and flow cytometry.

  9. Diagnostic tests based on human basophils: more potentials and perspectives than pitfalls.

    Science.gov (United States)

    de Weck, A L; Sanz, M L; Gamboa, P M; Aberer, W; Bienvenu, J; Blanca, M; Demoly, P; Ebo, D G; Mayorga, L; Monneret, G; Sainte-Laudy, J

    2008-01-01

    For the diagnosis of allergy, cellular basophil activation tests (BAT), e.g. histamine or sulfidoleukotriene release tests, have long been introduced, but the expression of basophil activation markers such as CD63 and CD203c detected by flow cytometry has attracted more recent attention. A recent opinion paper in this Journal has stressed not only the potential but also the possible pitfalls of flow-cytometric BAT. We have applied clinical validation of various BAT in various ways for several years, and our experience shows that these new technologies have more potentials and perspectives than pitfalls. A comprehensive review of clinically validated studies on allergy to aeroallergens, insect venoms, latex, food allergens and drugs, e.g. myorelaxants, beta-lactams, pyrazolones and non-steroidal anti-inflammatory drugs, as well as chronic urticaria shows clearly that even with different protocols, reproducible and meaningful results can be obtained. Although the available technologies may still be optimized and better standardized, there are no serious reasons to deprive allergic patients of clinically indicated BAT, which can be performed reliably by any laboratory with allergy and flow-cytometric capacity and expertise.

  10. [Basophilic line of the articular cartilage in normal and various pathological states].

    Science.gov (United States)

    Gongadze, L R

    1987-04-01

    Epiphyses of long tubular bones in the man and animals of various age, as well as experimental material of the adjuvant arthritis, with special reference to the basal part of the articular cartilage have been studied by means of histological, histochemical and histometrical methods. The structural-chemical organization of the basophilic line (tidemark) of the articular cartilage ensures its barrier role and participation in regulating selective permeability. Reconstruction of the tidemark in the process of physiological ageing and in cases of the articular pathology is aimed to preserve its integrity and in this way a complete differentiation of the noncalcified and calcified structures is secured. Disturbance of the basophilic line results in changes of the articular selective permeability, in invasion of vessels and structural elements of the bone marrow, and in development of profound distrophic and destructive changes of the cartilage--in deforming artrosis. Deflations in the structural-chemical organization of the tidemark indicate certain disturbances in the state of the system articular cartilage--subchondral bone. These data can be of prognostic importance.

  11. Elevated Toll-Like Receptor-Induced CXCL8 Secretion in Human Blood Basophils from Allergic Donors Is Independent of Toll-Like Receptor Expression Levels.

    Directory of Open Access Journals (Sweden)

    Markus Steiner

    Full Text Available Human blood basophils have recently gained interest in addition to their function as allergic effector cells. Previous work suggests the involvement of innate immune mechanisms in the development and exacerbation of allergic responses, which might be mediated by basophils. We assayed the expression levels of Toll-like receptor (TLR 1, 2, 4 and 6 on purified basophils from birch pollen-, house dust mite-, and non-allergic individuals. Additionally, we compared cytokine and chemokine secretion upon TLR stimulation in these basophil donor groups. Expression of TLR4 on the basophils of the allergic donor groups was decreased and CXCL8 secretion was elevated upon stimulation of TLR1/2 and TLR2/6 compared to the non-allergic donors. Decreased TLR expression and elevated CXCL8 secretion may represent possible mechanisms for aggravation of allergic symptoms in case of parasitic infection.

  12. iNKT Cells Are Responsible for the Apoptotic Reduction of Basophils That Mediate Th2 Immune Responses Elicited by Papain in Mice Following γPGA Stimulation.

    Science.gov (United States)

    Park, Hyun Jung; Lee, Sung Won; Park, Se-Ho; Hong, Seokmann

    2016-01-01

    Recent studies have demonstrated that Bacillus subtilis-derived poly-gamma glutamic acid (γPGA) treatment suppresses the development of allergic diseases such as atopic dermatitis (AD). Although basophils, an innate immune cell, are known to play critical roles in allergic immune responses and repeated long-term administration of γPGA results in decreased splenic basophils in an AD murine model, the underlying mechanisms by which γPGA regulates basophil frequency remain unclear. To investigate how γPGA modulates basophils, we employed basophil-mediated Th2 induction in vivo model elicited by the allergen papain protease. Repeated injection of γPGA reduced the abundance of basophils and their production of IL4 in mice, consistent with our previous study using NC/Nga AD model mice. The depletion of basophils by a single injection of γPGA was dependent on the TLR4/DC/IL12 axis. CD1d-dependent Vα14 TCR invariant natural killer T (iNKT) cells are known to regulate a variety of immune responses, such as allergy. Because iNKT cell activation is highly sensitive to IL12 produced by DCs, we evaluated whether the effect of γPGA on basophils is mediated by iNKT cell activation. We found that in vivo γPGA treatment did not induce the reduction of basophils in iNKT cell-deficient CD1d KO mice, suggesting the critical role of iNKT cells in γPGA-mediated basophil depletion at the early time points. Furthermore, increased apoptotic basophil reduction triggered by iNKT cells upon γPGA stimulation was mainly attributed to Th1 cytokines such as IFNγ and TNFα, consequently resulting in inhibition of papain-induced Th2 differentiation via diminishing basophil-derived IL4. Taken together, our results clearly demonstrate that γPGA-induced iNKT cell polarization toward the Th1 phenotype induces apoptotic basophil depletion, leading to the suppression of Th2 immune responses. Thus, elucidation of the crosstalk between innate immune cells will contribute to the design and

  13. Are basophil histamine release and high affinity IgE receptor expression involved in asymptomatic skin sensitization?

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Assing, K; Jensen, Lone Hummelshøj

    2006-01-01

    . However, a relationship between the AS status and FcepsilonRI has not been investigated. We aimed to characterize basophils from AS by looking at histamine release (HR) (sensitivity and reactivity) and the FcepsilonRI molecule, and compare it with nonatopic (NA) or allergic (A) persons....

  14. Role of IgG4 in histamine release from human basophil leucocytes. I. Sensitization of cells from normal donors

    DEFF Research Database (Denmark)

    Poulsen, L K; Stahl Skov, P; Mosbech, H

    1988-01-01

    Several conflicting reports on the ability of IgG4 to mediate type I allergic reactions have appeared lately. We have developed a model system for testing this possibility, using passive sensitization of basophil leucocytes from normal individuals. At first, the system was optimized with regard...

  15. Are basophil histamine release and high affinity IgE receptor expression involved in asymptomatic skin sensitization?

    DEFF Research Database (Denmark)

    Jensen, Bettina Margrethe; Assing, K; Jensen, Lone Hummelshøj;

    2006-01-01

    Immunoglobulin (Ig)E-sensitized persons with positive skin prick test, but no allergy symptoms, are classified as being asymptomatic skin sensitized (AS). The allergic type 1 disease is dependant on IgE binding to the high affinity IgE-receptor (FcepsilonRI) expressed on basophils and mast cells...

  16. Suppression of basophil histamine release and other IgE-dependent responses in childhood Schistosoma mansoni/hookworm coinfection

    DEFF Research Database (Denmark)

    Pinot de Moira, Angela; Fitzsimmons, Colin M; Jones, Frances M;

    2014-01-01

    in Ugandan schoolchildren coinfected with Schistosoma mansoni and hookworm. METHODS: Skin prick test (SPT) sensitivity to house dust mite allergen (HDM) and current wheeze were assessed pre-anthelmintic treatment. Nonspecific (anti-IgE), helminth-specific, and HDM-allergen-specific basophil histamine release...

  17. IL-33 induces IL-9 production in human CD4+ T cells and basophils

    DEFF Research Database (Denmark)

    Blom, Lars; Poulsen, Britta Cathrina; Jensen, Bettina M.

    2011-01-01

    IL-33, an IL-1 family member and ligand for the IL-1 receptor-related protein ST2, has been associated with induction of Th2 cytokines such as IL-4, IL-5, and IL-13. Here, we report that IL-33 can initiate IL-9 protein secretion in vitro in human CD4+ T cells and basophils isolated from peripheral...

  18. Brown Norway rat ovalbumin-specific immunoglobulin E antibodies increase the human basophil expression of CD63 marker

    NARCIS (Netherlands)

    Bellou, A.; Saint-Laudy, J.; Knippels, L.; Montémont, C.; Vauthier, E.; Gerard, P.; Pellegrom, H.; Koerkamp, E.K.; Lesesve, J.F.; Guéant, J.L.; Lambert, H.; Mallié, J.P.

    2003-01-01

    Anaphylactic shock is an immunoglobulin E (IgE)-dependent hypersensitivity. Biological tests like leucocyte histamine release (LHR) and human basophil activation (HBA), frequently used in human allergy, reflect both the amount of IgE fixed on cells and the cellular reactivity. To assess whether seru

  19. Basophil activation test in the diagnosis and monitoring of mastocytosis patients with wasp venom allergy on immunotherapy

    NARCIS (Netherlands)

    Bidad, Katayoon; Nawijn, Martijn C.; van Oosterhout, Antoon J. M.; van der Heide, Sicco; Oude Elberink, Joanne N. G.

    2014-01-01

    Background There is need for an accurate diagnostic test in mastocytosis patients with wasp venom allergy (WVA) and monitoring of these patients during immunotherapy (IT). In this study, we aimed to evaluate sensitivity and specificity of the Basophil Activation Test (BAT) as a diagnostic and monito

  20. Evaluation of the correlation and reproducibility between histamine, IL-4, and IL-13 release from human basophils.

    Directory of Open Access Journals (Sweden)

    Nahid Eskandari

    2014-06-01

    Full Text Available Human basophils play a key role in allergic diseases such as asthma and in a variety of immunological disorders. The generation of IL-4 and IL-13 can be induced from basophil by IgE-mediated and non-IgE-mediated mechanisms. Time and stimulus-dependent differences in the regulation of these cytokines could have relevance to their biological effects. The aim of the present study was activation of basophils in order to evaluate the extent of histamine, IL-4, and IL-13 generations. Basophil-enriched suspensions were prepared by Percoll gradients. The release of histamine and cytokines was assessed after activation with either anti-human IgE (1/1000 or 1/10000, 4 h or 24 h or IL-3 (100 ng/ ml, 24 h. Results were analysed statistically, using ANOVA test. Using anti-IgE, there was no significant correlation between the extent of either IL-4 (r=0.24, p=0.35 or IL-13 (r=0.47, p=0.098 and histamine release. Using IL-3 as stimulator, results showed that the extent of IL-13 correlated with histamine release(r=0.44, p=0.036. There was no correlation between the extent of IL-4 and the degree of either histamine (r=0.077, p=0.72 or IL-13 (r=0.162, p=0.5. The reproducibility of cytokines isolated from the same donor (on different occasions indicated that the ability of anti-IgE to induce cytokines was consistently similar for a given donor. Our data showed that the pathways leading to IL-3-triggering histamine release and IL-13 generation show similarity. Donor-dependent differences may be responsible for this wide range in the extent of releasibility. The ability of IL-3 to release cytokines from basophils showed a wider range.

  1. Evaluation of the correlation and reproducibility between histamine, IL-4, and IL-13 release from human basophils.

    Science.gov (United States)

    Eskandari, Nahid; Bastan, Reza; Ahmadi, Maryam; Peachell, Peter T

    2014-06-01

    Human basophils play a key role in allergic diseases such as asthma and in a variety of immunological disorders. The generation of IL-4 and IL-13 can be induced from basophil by IgE-mediated and non-IgE-mediated mechanisms. Time and stimulus-dependent differences in the regulation of these cytokines could have relevance to their biological effects. The aim of the present study was activation of basophils in order to evaluate the extent of histamine, IL-4, and IL-13 generations. Basophil-enriched suspensions were prepared by Percoll gradients. The release of histamine and cytokines was assessed after activation with either anti-human IgE (1/1000 or 1/10000, 4 h or 24 h) or IL-3 (100 ng/ ml, 24 h). Results were analysed statistically, using ANOVA test. Using anti-IgE, there was no significant correlation between the extent of either IL-4 (r=0.24, p=0.35) or IL-13 (r=0.47, p=0.098) and histamine release. Using IL-3 as stimulator, results showed that the extent of IL-13 correlated with histamine release(r=0.44, p=0.036). There was no correlation between the extent of IL-4 and the degree of either histamine (r=0.077, p=0.72) or IL-13 (r=0.162, p=0.5). The reproducibility of cytokines isolated from the same donor (on different occasions) indicated that the ability of anti-IgE to induce cytokines was consistently similar for a given donor. Our data showed that the pathways leading to IL-3-triggering histamine release and IL-13 generation show similarity. Donor-dependent differences may be responsible for this wide range in the extent of releasibility. The ability of IL-3 to release cytokines from basophils showed a wider range.

  2. Protein Fv produced during vital hepatitis is a novel activator of human basophils and mast cells.

    Science.gov (United States)

    Patella, V; Bouvet, J P; Marone, G

    1993-11-15

    Protein Fv is found in the normal liver and is released in the stools of patients suffering from viral hepatitis. Protein Fv isolated from five patients stimulated the release of histamine and sulfidopeptide leukotriene C4 from purified and unpurified peripheral blood basophils. Protein Fv absorbed with protein A-Sepharose coated with polyclonal IgG did not induce histamine secretion, whereas removal of putative contaminating Ig did not modify the releasing activity. The characteristics of the release reaction were similar to those of rabbit IgG anti-Fc fragment of human IgE (anti-IgE). There was an excellent correlation (Spearman rank coefficient (rs) = 0.83; p ADZ) blocked both anti-IgE- and protein Fv-induced releases, whereas human polyclonal IgG and a monoclonal IgG purified from another myeloma patient (patient ZEG) selectively blocked protein Fv-induced secretion. Protein Fv also induced the release of preformed (histamine and tryptase) and de novo synthesized mediators (sulfidopeptide leukotriene C4 and/or PGD2) from mast cells purified from human lung parenchyma and skin tissues. There was a significant correlation between the maximal percent histamine release induced by protein Fv and anti-IgE from skin mast cells (rs = 0.63; p < 0.01). There was also an excellent correlation between histamine and tryptase release caused by protein Fv from both lung (rs = 0.80; p < 0.001) and skin mast cells (rs = 0.70; p < 0.01). Thus, we established that protein Fv acts as a novel activator of human basophils and mast cells presumably by interacting with the VH domain of the IgE.

  3. Brown Norway rat ovalbumin-specific immunoglobulin E antibodies increase the human basophil expression of CD63 marker.

    Science.gov (United States)

    Bellou, A; Saint-Laudy, J; Knippels, L; Montémont, C; Vauthier, E; Gerard, P; Pellegrom, H; Koerkamp, E K; Lesesve, J F; Guéant, J L; Lambert, H; Mallié, J P

    2003-03-01

    Anaphylactic shock is an immunoglobulin E (IgE)-dependent hypersensitivity. Biological tests like leucocyte histamine release (LHR) and human basophil activation (HBA), frequently used in human allergy, reflect both the amount of IgE fixed on cells and the cellular reactivity. To assess whether serum-specific IgE from Brown Norway (BN) rats prepared for ovalbumin (OVA)-induced anaphylactic shocks can activate human basophils which has a potential interest in experimental allergy: such a test could rapidly assert an IgE sensitization in laboratory animals genetically T-helper 2 (Th2)-predisposed. Rats (n = 39) were immunized three times (day 0, day 5 and day 21) with OVA injected subcutaneously. One week after the third immunization, a shock was induced with an intravenous (i.v.) bolus of OVA. Sensitization was assessed by passive cutaneous anaphylaxis (PCA) test and dosages of serum IgE antibodies anti-OVA by enzyme-linked immunosorbent assay. Blood basophils were counted before and during the shock. Before the shock induction (at day 21), an LHR test was performed on rat blood, and human basophils were sensitized with rat sera. HBA was demonstrated by the increase in the percentage of cells expressing CD63 antigen membrane, measured by flow cytometry. Twenty-one days after the first subcutaneous (s.c.) immunization, the rat serum induced a significant HBA. HBA was observed neither with the same serum previously heated nor with the serum from nonimmunized rats (NIRs). OVA-specific IgEs were significantly increased in immunized rat (IR) serum. The PCA test was negative when the serum was previously heated (56 degrees C). We never observed any circulating basophils, and LHR test was negative. After OVA i.v. administration, all IRs died rapidly. HBA testing strongly suggests a mediation by specific IgE in the increase of CD63 in BN rats. Thus, HBA test seems useful in assessing whether an experimental allergy was induced in animals genetically predisposed to an immune

  4. Validation of basophil histamine release against the autologous serum skin test and outcome of serum-induced basophil histamine release studies in a large population of chronic urticaria patients

    DEFF Research Database (Denmark)

    Platzer, M H; Grattan, C E H; Poulsen, Lars K.

    2005-01-01

    Endogenous histamine-releasing factors (HRFs) are involved in 30-60% of patients with chronic urticaria (CU). Evidence for their existence comes from in vivo studies of autoreactivity with the autologous serum skin test (ASST), in vitro immunoassays demonstrating autoantibodies against the immuno......Endogenous histamine-releasing factors (HRFs) are involved in 30-60% of patients with chronic urticaria (CU). Evidence for their existence comes from in vivo studies of autoreactivity with the autologous serum skin test (ASST), in vitro immunoassays demonstrating autoantibodies against...... the immunoglobulin E (IgE) or the high affinity IgE receptor (FcepsilonRI) and serum-induced histamine release (HR) from basophils and mast cells. We have examined the correlation between the ASST and a new basophil histamine-releasing assay (the HR-Urtikaria test) in a group of well-characterized CU patients...

  5. Screening anaphylactic components of MaiLuoNing injection by using rat basophilic leukemia-2H3 cell membrane chromatography coupled with HPLC-ESI-TOF-MS.

    Science.gov (United States)

    Han, Shengli; Lv, Yanni; Xue, Wenjing; Cao, Jiao; Cui, Ronghua; Zhang, Tao

    2016-02-01

    MaiLuoNing injection is a traditional Chinese medicine that used clinically since the 1950s in China. However, anaphylactic reactions, through the potentiation of mast cell degranulation, have been reported. In the present study, a rat basophilic leukemia-2H3 cell membrane chromatography coupled with high-performance liquid chromatography and electrospray ionization-ion trap-time of flight-mass spectrometry method was established for screening, analyzing, and identifying the potential anaphylactic components of MaiLuoNing injection. Harpagoside, a potential degranulator of rat basophilic leukemia-2H3 cells, was retained in rat basophilic leukemia-2H3 cell membrane chromatography. We aimed to evaluate the retained components to determine which of those were capable of inducing degranulation of basophilic leukemia cells. A β-hexosaminidase assay revealed that harpagoside can induce rat basophilic leukemia-2H3 cell degranulation in a dose-dependent manner. BLBA/c mice also exhibit passive cutaneous anaphylaxis in response to harpagoside. These results indicate that rat basophilic leukemia-2H3 cell membrane chromatography coupled with high-performance liquid chromatography and electrospray ionization ion trap time-of-flight mass spectrometry is effective in screening for the anaphylactic components of MaiLuoNing injection.

  6. Membrane and cytoskeletal changes associated with IgE-mediated serotonin release from rat basophilic leukemia cells

    OpenAIRE

    1985-01-01

    Binding of antigen to IgE-receptor complexes on the surface of RBL-2H3 rat basophilic leukemia cells is the first event leading to the release of cellular serotonin, histamine, and other mediators of allergic, asthmatic, and inflammatory responses. We have used dinitrophenol- conjugated bovine serum albumin (DNP-BSA) as well as the fluorescent antigen, DNP-B-phycoerythrin, and the electron-dense antigen, DNP-BSA- gold, to investigate dynamic membrane and cytoskeletal events associated with th...

  7. A positive serum basophil histamine release assay is a marker for ciclosporin-responsiveness in patients with chronic spontaneous urticaria

    DEFF Research Database (Denmark)

    Iqbal, Kamran; Bhargava, Kapil; Skov, Per Stahl;

    2012-01-01

    ABSTRACT: The electronic records of 398 patients with chronic spontaneous urticaria (CSU) who had had a serum basophil histamine release assay (BHRA) performed as a marker of functional autoantibodies were audited. The BHRA was positive in 105 patients (26.4%). Fifty eight were treated with ciclo...... with ciclosporin because they were H1 anti-histamine unresponsive. CSU patients with a positive BHRA were more likely to respond clinically (P...

  8. [Level of histamine in supernatants from the basophil activation test: applications to hymenoptera allergy and drug allergy--preliminary study].

    Science.gov (United States)

    Tamim, S; Lauret, M G; Drouet, M; Sabbah, A

    1999-02-01

    Histamine Release technic consists in calculating histamine liberated by blood cells in touch with an allergen. To this day, this method is only used in Hymenoptera venom allergy diagnosis. The principle of this study is to measure histamine released by activated basophils in surnageons of Basophil Activation Test (BAT) for different allergens: Hymenoptera venoms: Bee, White Faced Hornet, Vespula Wasp. Drugs: Cefaperos, Clamoxyl, Alfatil, Rapifen, Diprivan, Nesdonal, Mivacron. A threshold of positivity (amplification factor in comparison with the control) is determined for these two classes of allergens: 45 for Hymenoptera venoms and 9 for drugs. These results, compared to the other diagnosis technics (Histamine Release, Basophil Activation Test, Prick Tests) discloses very high correlation rates in each case. This method seems to be a reliable method for Hymenoptera venom allergy diagnosis and for drugs allergy diagnosis too. However, this study is based on a few number of patients, so a significant statistic conclusion can't be expressed but it opens an interesting way of research.

  9. Detection of homocytotropic antibody in lambs infested with the louse, Bovicola ovis, using a basophil histamine-release assay.

    Science.gov (United States)

    Pfeffer, A; Phegan, M D; Bany, J

    1997-07-01

    The utility of a basophil histamine-release assay using washed whole blood cells was examined in lambs and was used to determine if homocytotropic antibody with specificity for Bovicola ovis was produced in response to infestation with the louse. Maximal histamine release in the assay in response to Concanavalin A, anti-ovine IgE monoclonal antibody and, in sensitized lambs, to B. ovis antigen ranged from 18 to 48%. Histamine release from blood cells in response to B. ovis antigen was significantly higher in louse-infested lambs than in louse-naive lambs and was significantly correlated with louse and cockle scores. Passive cutaneous anaphylaxis (PCA) tests were negative with sera obtained from the lambs at the same time as blood for the basophil histamine-release assay. Serum histamine levels also were significantly higher in the louse-infested lambs than in louse-naive lambs and were significantly correlated with louse and cockle scores. The present results support a role for B. ovis-specific homocytotropic antibody in the development of cockle and indicate that the basophil histamine-release assay is more sensitive than the PCA test.

  10. Effects of Echium plantagineum L. Bee Pollen on Basophil Degranulation: Relationship with Metabolic Profile

    Directory of Open Access Journals (Sweden)

    Eduarda Moita

    2014-07-01

    Full Text Available This study aimed to evaluate the anti-allergic potential of Echium plantagineum L. bee pollen and to characterize its primary metabolites. The activity of E. plantagineum hydromethanolic extract, devoid of alkaloids, was tested against β-hexosaminidase release in rat basophilic leukemic cells (RBL-2H3. Two different stimuli were used: calcium ionophore A23187 and IgE/antigen. Lipoxygenase inhibitory activity was evaluated in a cell-free system using soybean lipoxygenase. Additionally, the extract was analysed by HPLC-UV for organic acids and by GC-IT/MS for fatty acids. In RBL-2H3 cells stimulated either with calcium ionophore or IgE/antigen, the hydromethanolic extract significantly decreased β-hexosaminidase release until the concentration of 2.08 mg/mL, without compromising cellular viability. No effect was found on lipoxygenase. Concerning extract composition, eight organic acids and five fatty acids were determined for the first time. Malonic acid (80% and α-linolenic acid (27% were the main compounds in each class. Overall, this study shows promising results, substantiating for the first time the utility of intake of E. plantagineum bee pollen to prevent allergy and ameliorate allergy symptoms, although a potentiation of an allergic response can occur, depending on the dose used.

  11. In vitro stimulation with a strongly pulsed electromagnetic field on rat basophilic leukemia cells

    Science.gov (United States)

    Choi, J. W.; Shin, S. C.; Kim, S.; Chung, E. R.; Bang, J. H.; Cho, G. I.; Choi, S. D.; Park, Y. S.; Jang, T. S.; Yoo, Y. M.; Lee, S. S.; Hwang, D. G.

    2010-05-01

    In this study, the effects of pulsed electromagnetic field stimulation with a strong magnetic field on rat basophilic leukemia (RBL-2H3) cells were investigated to confirm the efficacy of the magnetic stimulator for biomedical applications. The maximum intensity of the magnetic field generated from the stimulation coil was 0.203 T, and the transition time was 126 μs. The oscillation time and frequency of the pulsed field were almost 0.1 ms and 8 kHz, respectively. The cell count as well as the mRNA expression and DNA sequence of the cytokine genes, such as the tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4), of the stimulated RBL-2H3 cells were analyzed with a hemocytometer and via reverse transcriptase polymerase chain reaction to determine the physiological response under a strong pulse field. After 12 h stimulation, cell death was observed at an increasing scale with the increase in the stimulation time. On the other hand, the cells that were stimulated for 10 min almost doubled as the interval time between the stimulations was extended.

  12. Serum Specific IgE to Thyroid Peroxidase Activates Basophils in Aspirin Intolerant Urticaria.

    Science.gov (United States)

    Shin, Yoo Seob; Suh, Dong-Hyeon; Yang, Eun-Mi; Ye, Young-Min; Park, Hae-Sim

    2015-06-01

    Thyroid antibodies are frequently observed in urticaria patients, but their roles in urticaria are not clearly elucidated. We investigated the role of serum specific IgE to thyroid peroxidase (TPO) in patients with aspirin intolerant acute urticaria (AIAU) and aspirin intolerant chronic urticaria (AICU). We recruited 59 AIAU and 96 AICU patients with 69 normal controls (NC). Serum specific IgE to TPO was measured by manual direct ELISA, and CD203c expressions on basophil with additions of TPO were measured to prove a direct role of TPO in effector cells. The prevalences of serum specific IgE to TPO were significantly higher in AIAU (15.2%) and AICU groups (7.5%) compared to NC (0%, P=0.018: P=0.013, respectively). Flow cytometry showed CD203c induction in a dose dependent manner with serial additions of TPO in some AIAU and AICU patients having high specific IgE to TPO. Our findings show that the prevalence of serum specific IgE to TPO was significantly higher in both AIAU and AICU patients than in NC. It is suggested that specific IgE to TPO play a pathogenic role in AIAU and AICU.

  13. Production of cytokines of type II (IL-4 and IL-13) by basophil polynuclear: effects of chimiokins and stimulation by the diesel particulates; Production de cytokines de type 2 (IL-4 et IL-13) par les polynucleaires basophiles: effects des chimiokines et stimulation par les particules de diesel

    Energy Technology Data Exchange (ETDEWEB)

    Devouassoux, G.

    2000-07-01

    Cytokines of type II are critical factors in the generation of allergic inflammation. There is a correlation between the allergy incidence and the pollutants exposition. The impact of the diesel particles on the allergic response is established but their impact on the basophil is unknown. This study shows that the basophil is a target for the diesel particles, responsible of an important production of IL-4, independent of the clinical status but dependent of the oxidative stress. (A.L.B.)

  14. In vitro histamine release from basophils of asthmatic and atopic individuals in D/sub 2/O

    Energy Technology Data Exchange (ETDEWEB)

    Tung, R.; Lichtenstein, L.M.

    1982-05-01

    It was found that spontaneous histamine release from human basophils in H/sub 2/O-based buffers is negligible; in D/sub 2/O-based buffers, however, release is observed with the cells of some donors. Analysis of this phenomenon revealed release from the basophils of 1 of 22 control individuals (5%), 15 of 47 patients with allergic rhinitis (32%), and 14 of 20 asthmatic patients (70%). The difference between both patient groups and controls and between atopics and asthmatics was highly significant. That D/sub 2/O release was not cytotoxic is suggested by the finding that 37/sup 0/ was optimal, with inhibition at 4/sup 0/C or 46/sup 0/C as well as by EDTA, 2-deoxyglucose, and dibromoacetophenone, an inhibitor of phospholipase A/sub 2/. The release mechanism was unusual in that dibutyryl cAMP and agonists that cause an increase in cAMP lead to no inhibition. No correlation was noted between the total serum IgE level (and thus the number of IgE receptors on the basophil surface) and the magnitude of D/sub 2/O release. No increase in D/sub 2/O release was observed in 17 ragweed-sensitive patients through a ragweed season. A unique property of D/sub 2/O release was the loss of reactivity by preincubating cells at 37/sup 0/C for 30 min before adding D/sub 2/O. Non-D/sub 2/O-reactive cells could be ''converted'' to D/sub 2/O-reactive cells by incubation with antigen in the whole blood phase during leukocyte isolation; these cells showed the same loss of releaseability at 37/sup 0/C and an inhibitor profile similar to D/sub 2/O-responsive cells from ragweed allergic or asthmatic patients. We suggest that D/sub 2/O-based buffers reveal, in atopic and asthmatic patients, in vivo basophil activation; whether this is due to IgE cross-links, to C split products, or to other stimuli is not yet clear.

  15. Comparison of CD63 Upregulation Induced by NSAIDs on Basophils and Monocytes in Patients with NSAID Hypersensitivity

    Directory of Open Access Journals (Sweden)

    N. Abuaf

    2012-01-01

    Full Text Available Background. An in vitro basophil activation test, based on the detection of CD63 upregulation induced by NSAIDs, has been described. Its clinical significance remains controversial. Objectives. In patients with a history of nonallergic NSAID hypersensitivity, stratified according to the severity of the symptoms, to assess with NSAIDs the predictive value of basophil (BAT and monocyte (MAT activation tests. Patients/Methods. Sixty patients who had NSAIDs-induced or exacerbated urticaria/angiooedema and 20 controls was included. After incubation with NSAIDs or acetaminophen, leukocytes were analysed for CD63 upregulation. Results. With aspirin, the sensitivity (37% and specificity (90% of BAT agree with already published results. In contrast, when patients had had cutaneous and visceral reactions, the frequency of positive BAT 14/22 (64%, P<0.001 or MAT 10/22 (46%, P<0.01 were increased. Conclusions. Positive tests were more frequent among patients having a severe hypersensitivity contrasting with the other patients who had results similar to controls.

  16. A Crystallin Fold in the Interleukin-4-inducing Principle of Schistosoma mansoni Eggs (IPSE/α-1) Mediates IgE Binding for Antigen-independent Basophil Activation.

    Science.gov (United States)

    Meyer, N Helge; Mayerhofer, Hubert; Tripsianes, Konstantinos; Blindow, Silke; Barths, Daniela; Mewes, Astrid; Weimar, Thomas; Köhli, Thies; Bade, Steffen; Madl, Tobias; Frey, Andreas; Haas, Helmut; Mueller-Dieckmann, Jochen; Sattler, Michael; Schramm, Gabriele

    2015-09-04

    The IL-4-inducing principle from Schistosoma mansoni eggs (IPSE/α-1), the major secretory product of eggs from the parasitic worm S. mansoni, efficiently triggers basophils to release the immunomodulatory key cytokine interleukin-4. Activation by IPSE/α-1 requires the presence of IgE on the basophils, but the detailed molecular mechanism underlying activation is unknown. NMR and crystallographic analysis of IPSEΔNLS, a monomeric IPSE/α-1 mutant, revealed that IPSE/α-1 is a new member of the βγ-crystallin superfamily. We demonstrate that this molecule is a general immunoglobulin-binding factor with highest affinity for IgE. NMR binding studies of IPSEΔNLS with the 180-kDa molecule IgE identified a large positively charged binding surface that includes a flexible loop, which is unique to the IPSE/α-1 crystallin fold. Mutational analysis of amino acids in the binding interface showed that residues contributing to IgE binding are important for IgE-dependent activation of basophils. As IPSE/α-1 is unable to cross-link IgE, we propose that this molecule, by taking advantage of its unique IgE-binding crystallin fold, activates basophils by a novel, cross-linking-independent mechanism.

  17. Antiallergic tea catechin, (-)-epigallocatechin-3-O-(3-O-methyl)-gallate, suppresses FcepsilonRI expression in human basophilic KU812 cells.

    Science.gov (United States)

    Fujimura, Yoshinori; Tachibana, Hirofumi; Maeda-Yamamoto, Mari; Miyase, Toshio; Sano, Mitsuaki; Yamada, Koji

    2002-09-25

    We previously found that the O-methylated derivative of (-)-epigallocatechin-3-O-gallate (EGCg), (-)-epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG' '3Me), has potent antiallergic activity. The high-affinity IgE receptor, FcepsilonRI, is found at high levels on basophils and mast cells and plays a key role in a series of acute and chronic human allergic reactions. To understand the mechanism of action for the antiallergic EGCG' '3Me, the effect of EGCG' '3Me on the cell surface expression of FcepsilonRI in human basophilic KU812 cells was examined. Flow cytometric analysis showed that EGCG' '3Me was able to decrease the cell surface expression of FcepsilonRI. Moreover, immunoblot analysis revealed that total cellular expression of the FcepsilonRI alpha chain decreased upon treatment with EGCG' '3Me. FcepsilonRI is a tetrameric structure comprising one alpha chain, one beta chain, and two gamma chains. The level of mRNA production of each subunit in KU812 cells was investigated. EGCG' '3Me reduced FcepsilonRI alpha and gamma mRNA levels. The cross-linkage of FcepsilonRI causes the activation of basophils, which leads to the secretion of inflammatory mediators including histamine. EGCG' '3Me treatment inhibited the FcepsilonRI cross-linking-induced histamine release. These results suggested that EGCG' '3Me can negatively regulate basophil activation through the suppression of FcepsilonRI expression.

  18. IgE in the absence of allergen induces the expression of monocyte chemoattractant protein-1 in the rat basophilic cell-line RBL-2H3.

    Science.gov (United States)

    Ahn, Ki Bum; Jeon, Jun Ho; Kang, Seok-Seong; Chung, Dae Kyun; Yun, Cheol-Heui; Han, Seung Hyun

    2014-11-01

    Recently, basophils have been suggested to produce inflammatory mediators in response to IgE in the absence of allergens. Monocyte chemoattractant protein-1 (MCP-1) plays an important role in the initiation of inflammatory responses by recruiting various immune cells to the site of allergic inflammation. In the present study, we examined whether IgE under allergen-free conditions could stimulate basophils and lead to the production of MCP-1. Exposure of the rat basophilic cell-line RBL-2H3 to IgE without allergen resulted in a dose- and time-dependent induction of MCP-1 expression at both the mRNA and protein level. Although allergen was not necessary for IgE-induced MCP-1 expression, it was essential for degranulation as determined by β-hexosaminidase release assay. IgE enhanced phosphorylation of MAP kinases including ERK, p38 kinase, and JNK. However, IgE-induced MCP-1 expression was attenuated by inhibitors for JNK and PKC. Concomitantly, IgE induced activation of AP-1, which is an important transcription factor for MCP-1 gene expression in RBL-2H3 cells. Taken together, our results suggest that IgE alone is sufficient to stimulate basophils to increase expression of MCP-1, which in turn might contribute to the inflammatory response.

  19. Flow cytometric analysis of the inhibition of human basophil activation by histamine high dilutions – a replication study

    Directory of Open Access Journals (Sweden)

    Chantal Wälchli

    2012-09-01

    Full Text Available Background: Inhibition of human basophil activation by highly diluted histamine was reported to be a reliable experimental model to examine biological effects of high dilutions. However, independent replications did not always yield concordant results. Aims: We aimed at performing an independent replication of a former study [1] using rigorously controlled experimental conditions to minimise confounding factors. Materials and Methods: In 20 independent experiments, human basophils were treated with highly diluted histamine (15cH, 16cH, corresponding to 10-30-10-32 M prior to activation by fMLP (formyl-methionyl-leucyl-phenylalanine peptide. Controls were treated with analogously diluted water (15cH, 16cH. The dilutions were prepared freshly for each experiment in deionised water by successive steps of centesimal dilution and agitation (10 s vortex at high speed. Highly diluted samples were blinded and randomised. All samples were set in triplicates. Activated basophils were determined by flow cytometry using anti-CD203c. 20 independent systematic negative control (SNC experiments were carried out to investigate possible systematic errors. Results: No difference in basophil activation was observed between the highly diluted histamine samples and the highly diluted water controls. There was no evidence for a blood donor specificity of the results. The SNC experiments demonstrated the stability of the test system. Experimental variability within and between experiments was slightly reduced for the highly diluted histamine samples. Discussion: This study was designed as an independent reproduction of a former study [1]. Though we strictly adopted the experimental procedure described in [1], our results do not confirm the large inhibitory effects observed for histamine 15cH and 16cH. This lack of reproducibility might be due to minor differences in the experimental design, such as blinding and randomising of the samples, which we chose to

  20. Suppression of Basophil Histamine Release and Other IgE-dependent Responses in Childhood Schistosoma mansoni/hookworm Coinfection

    Science.gov (United States)

    Pinot de Moira, Angela; Fitzsimmons, Colin M.; Jones, Frances M.; Wilson, Shona; Cahen, Pierre; Tukahebwa, Edridah; Mpairwe, Harriet; Mwatha, Joseph K.; Bethony, Jeffrey M.; Skov, Per S.; Kabatereine, Narcis B.; Dunne, David W.

    2014-01-01

    Background. The poor correlation between allergen-specific immunoglobulin E (asIgE) and clinical signs of allergy in helminth infected populations suggests that helminth infections could protect against allergy by uncoupling asIgE from its effector mechanisms. We investigated this hypothesis in Ugandan schoolchildren coinfected with Schistosoma mansoni and hookworm. Methods. Skin prick test (SPT) sensitivity to house dust mite allergen (HDM) and current wheeze were assessed pre-anthelmintic treatment. Nonspecific (anti-IgE), helminth-specific, and HDM-allergen-specific basophil histamine release (HR), plus helminth- and HDM-specific IgE and IgG4 responses were measured pre- and post-treatment. Results. Nonspecific- and helminth-specific-HR, and associations between helminth-specific IgE and helminth-specific HR increased post-treatment. Hookworm infection appeared to modify the relationship between circulating levels of HDM-IgE and HR: a significant positive association was observed among children without detectable hookworm infection, but no association was observed among infected children. In addition, hookworm infection was associated with a significantly reduced risk of wheeze, and IgG4 to somatic adult hookworm antigen with a reduced risk of HDM-SPT sensitivity. There was no evidence for S. mansoni infection having a similar suppressive effect on HDM-HR or symptoms of allergy. Conclusions. Basophil responsiveness appears suppressed during chronic helminth infection; at least in hookworm infection, this suppression may protect against allergy. PMID:24782451

  1. Autologous serum skin test reactivity and basophil histamine release test in patients with nasal polyposis: preliminary results.

    Science.gov (United States)

    Zambetti, G; Ciofalo, A; Soldo, P; Fusconi, M; Romeo, R; Greco, A; Altissimi, G; Macri, G F; Marinelli, C; Pagliuca, G; De Vincentiis, M

    2010-01-01

    An eosinophilic inflammatory process is generally observed in patients suffering from nasal polyposis (NP), however its onset has not yet been defined. It has been suggested that immune activation of inflammatory cells may be the cause. The aim of this study is to verify whether autoantibodies and/or histamine-releasing factors are present in the serum of patients suffering from NP. In fact, we assume that autoantibodies and/or histamine-releasing factors, as already demonstrated in chronic idiopathic urticaria and asthma, may be involved in the pathogenesis of NP. In this case-control analytical study 40 patients with NP and 27 control subjects underwent the in vivo autologous serum skin test (ASST). The sera from 6 patients suffering from NP and 9 control group subjects, who had all been previously studied and randomly selected, underwent basophil histamine release assay from normal donor as a pilot study. The ASST showed positive results in 55% of patients suffering from NP versus 8% of the control group (p= .00006), the basophil histamine release test (BHRT) turned out positive in all patients tested and in 11% of the control group. We found a weak positive correlation between the percentage of histamine release and the wheal diameter. ASST reactivity is very frequent in patients suffering from NP, thus suggesting the presence of histamine-releasing factors in the blood stream. The BHRT was positive in the serum of all patients, thus suggesting the presence of anti-FcepsilonRI, anti-IgE autoantibodies and/or other histamine-releasing factors, the presence of which can play a role in triggering and maintaining the eosinophilic inflammatory process in NP.

  2. Inhibition of basophil histamine release by gangliosides. Further studies on the significance of cell membrane sialic acid in the histamine release process

    DEFF Research Database (Denmark)

    Jensen, C; Norn, S; Thastrup, Ole

    1987-01-01

    Histamine release from human basophils was inhibited by preincubation of the cells with a glucolipid mixture containing sialic acid-containing gangliosides. This was true for histamine release induced by anti-IgE, Concanavalin A and the calcium ionophore A23187, whereas the release induced by S...... with the glucolipid mixture increased the sialic acid content of the cells, and this increase was attributed to an insertion of gangliosides into the cell membrane. The inhibition of histamine release was abolished by increasing the calcium concentration, which substantiates our previous findings that cell membrane...... sialic acid in basophil leukocytes is involved in the regulation of histamine release, possibly by a modulation of the transmembraneous calcium fluxes preceding histamine release....

  3. Screening antiallergic components from Carthamus tinctorius using rat basophilic leukemia 2H3 cell membrane chromatography combined with high-performance liquid chromatography and tandem mass spectrometry.

    Science.gov (United States)

    Han, Shengli; Huang, Jing; Cui, Ronghua; Zhang, Tao

    2015-02-01

    Carthamus tinctorius, used in traditional Chinese medicine, has many pharmacological effects, such as anticoagulant effects, antioxidant effects, antiaging effects, regulation of gene expression, and antitumor effects. However, there is no report on the antiallergic effects of the components in C. tinctorius. In the present study, we investigated the antiallergic components of C. tinctorius and its mechanism of action. A rat basophilic leukemia 2H3/cell membrane chromatography coupled online with high-performance liquid chromatography and tandem mass spectrometry method was developed to screen antiallergic components from C. tinctorius. The screening results showed that Hydroxysafflor yellow A, from C. tinctorius, was the targeted component that retained on the rat basophilic leukemia 2H3/cell membrane chromatography column. We measured the amount of β-hexosaminidase and histamine released in mast cells and the key markers of degranulation. The release assays showed that Hydroxysafflor yellow A could attenuate the immunoglobulin E induced release of allergic cytokines without affecting cell viability from 1.0 to 50.0 μM. In conclusion, the established rat basophilic leukemia 2H3 cell membrane chromatography coupled with online high-performance liquid chromatography and tandem mass spectrometry method successfully screened and identified Hydroxysafflor yellow A from C. tinctorius as a potential antiallergic component. Pharmacological analysis elucidated that Hydroxysafflor yellow A is an effective natural component for inhibiting immunoglobulin E-antigen-mediated degranulation.

  4. Structural analysis of the endogenous glycoallergen Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis and its recognition by human basophils.

    Science.gov (United States)

    Rodríguez-Romero, Adela; Hernández-Santoyo, Alejandra; Fuentes-Silva, Deyanira; Palomares, Laura A; Muñoz-Cruz, Samira; Yépez-Mulia, Lilian; Orozco-Martínez, Socorro

    2014-02-01

    Endogenous glycosylated Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis is an important latex allergen that is recognized by IgE antibodies from patients who suffer from latex allergy. The carbohydrate moieties of Hev b 2 constitute a potentially important IgE-binding epitope that could be responsible for its cross-reactivity. Here, the structure of the endogenous isoform II of Hev b 2 that exhibits three post-translational modifications, including an N-terminal pyroglutamate and two glycosylation sites at Asn27 and at Asn314, is reported from two crystal polymorphs. These modifications form a patch on the surface of the molecule that is proposed to be one of the binding sites for IgE. A structure is also proposed for the most important N-glycan present in this protein as determined by digestion with specific enzymes. To analyze the role of the carbohydrate moieties in IgE antibody binding and in human basophil activation, the glycoallergen was enzymatically deglycosylated and evaluated. Time-lapse automated video microscopy of basophils stimulated with glycosylated Hev b 2 revealed basophil activation and degranulation. Immunological studies suggested that carbohydrates on Hev b 2 represent an allergenic IgE epitope. In addition, a dimer was found in each asymmetric unit that may reflect a regulatory mechanism of this plant defence protein.

  5. The effects of thermal stimuli on intracellular calcium change and histamine releases in rat basophilic leukemia mast cells

    Science.gov (United States)

    Wu, Zu-Hui; Zhu, Dan; Chen, Ji-Yao; Zhou, Lu-Wei

    2012-05-01

    The effects of thermal stimuli on rat basophilic leukemia mast cells were studied. The cells in calcium-contained or calcium-free buffers were thermally stimulated in the temperature range of 25-60 °C. The corresponding calcium ion concentration in cells [Ca2+]i as well as the released histamine from cells was measured with fluorescence staining methods. The ruthenium red (RR), a block of membrane calcium channels (transient receptor potential family V (TRPV)), was used in experiments. Under the stimulus of 25-50 °C, no significant difference on [Ca2+]i was found between these three groups of the cells in calcium-contained buffer without or with RR and cells in calcium-free saline, indicating that the increased calcium in cytosol did not result from the extracellular buffer but came from the intracellular calcium stores. The [Ca2+]i continuously increased under the temperature of 50-60 °C, but the RR and calcium-free saline can obviously diminish the [Ca2+]i increase at these high temperatures, reflecting that the opening of the TRPV2 channels leads to a calcium influx resulting in the [Ca2+]i increment. The histamine release also became significant in these cases. Since the released histamine is a well-known mediator for the microcirculation promotion, the histamine release from mast cells could be one of the mechanisms of thermal therapy.

  6. The LINA Study: Higher Sensitivity of Infant Compared to Maternal Eosinophil/Basophil Progenitors to Indoor Chemical Exposures

    Directory of Open Access Journals (Sweden)

    Friederike Hörnig

    2016-01-01

    Full Text Available Purpose. Enhanced eosinophil/basophil (Eo/B progenitor cell levels are known to be associated with allergic inflammation and atopy risk. The aim of the present study was to investigate the influence of different indoor exposures on the recruitment and differentiation of Eo/B progenitors in mother-child pairs. Methods. In 68 mother-child pairs of the LINA study peripheral blood mononuclear cells were used to assess Eo/B colony forming units (CFUs. Information about disease outcomes and indoor exposures was obtained from questionnaires. Indoor concentrations of volatile organic compounds (VOCs were measured by passive sampling. Results. Infant’s Eo/B CFUs were positively associated with exposure to tobacco smoke, disinfectants, or VOCs. In contrast, for maternal Eo/B CFUs, only a few associations were seen. Higher numbers of infant Eo/B CFUs were observed in children with wheezing symptoms within the second year of life. Conclusions. We demonstrate that infant’s hematopoietic cells seem to respond with more sensitivity to environmental exposure compared to maternal cells. At least in infants, an activation of these hematopoietic cells by environmental exposure could contribute to an enhanced risk for the development of respiratory outcomes.

  7. Distribution and dynamics of rat basophilic leukemia immunoglobulin E receptors (FcepsilonRI) on planar ligand-presenting surfaces.

    Science.gov (United States)

    Spendier, Kathrin; Carroll-Portillo, Amanda; Lidke, Keith A; Wilson, Bridget S; Timlin, Jerilyn A; Thomas, James L

    2010-07-21

    There is considerable interest in the signaling mechanisms of immunoreceptors, especially when triggered with membrane-bound ligands. We have quantified the spatiotemporal dynamics of the redistribution of immunoglobulin E-loaded receptors (IgE-FcepsilonRI) on rat basophilic leukemia-2H3 mast cells in contact with fluid and gel-phase membranes displaying ligands for immunoglobulin E, using total internal reflection fluorescence microscopy. To clearly separate the kinetics of receptor redistribution from cell spreading, and to precisely define the initial contact time (+/-50 ms), micropipette cell manipulation was used to bring individual cells into contact with surfaces. On ligand-free surfaces, there are micron-scale heterogeneities in fluorescence that likely reflect regions of the cell that are more closely apposed to the substrate. When ligands are present, receptor clusters form with this same size scale. The initial rate of accumulation of receptors into the clusters is consistent with diffusion-limited trapping with D approximately 10(-1) microm2/s. These results support the hypothesis that clusters form by diffusion to cell-surface contact regions. Over longer timescales (>10 s), individual clusters moved with both diffusive and directed motion components. The dynamics of the cluster motion is similar to the dynamics of membrane fluctuations of cells on ligand-free fluid membranes. Thus, the same cellular machinery may be responsible for both processes.

  8. Biochemical and phenotypic characterization of human basophilic cells derived from dispersed fetal liver with murine T cell factors

    Energy Technology Data Exchange (ETDEWEB)

    Seldin, D.C.; Caulfield, J.P.; Hein, A.; Osathanondh, R.; Nabel, G.; Schlossman, S.F.; Stevens, R.L.; Austen, K.F.

    1986-03-15

    Metachromatically granulated cells were generated from human fetal liver stem cells cultured in heterologous mouse conditioned medium rich in interleukin 3. After 2 to 3 wk of culture with biweekly changes of medium and selection of nonadherent cells, all cells present in five cultures had cytoplasmic granules. Ultrastructurally, many granules contained fibrillar material or electron-dense cores with fibrils and vesicular fragments. In addition, the granules of many cells were filled with electron-dense material, which in some cases had a fine structure consisting of concentric whorls or a reticular pattern. Analysis of high-affinity IgE receptors on the cultured cells by flow cytometry demonstrated a unimodel fluorescence pattern, suggesting that most cells were in the basophil or mast cell lineage. The cells contained 52 ng/10/sup 6/ cells of histamine and incorporated (/sup 35/S)sulfate at an average rate of 31,300 cpm/10/sup 6/ cells/4 hr into 175,000 m.w. chondroitin sulfate A proteoglycans. Upon activation with 1 ..mu..M calcium ionophore A23187, the cultured cells released 53% of their cell-associated histamine and metabolized arachidonic acid to 15.0 ng/10/sup 6/ cells of immunoreactive leukotriene C/sub 4/ equivalents, 0.5 ng/10/sup 6/ cells of leukotriene B/sub 4/, and 3.1 ng/10/sup 6/ cells of prostaglandin D/sub 2/ (means, n = 3). Thus, stem cells present in human fetal liver give rise, as do stem cells in mouse fetal liver, to metachromatically granulated cells when cultured in the presence of mouse interleukin 3.

  9. Structural analysis of the endogenous glycoallergen Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis and its recognition by human basophils

    Energy Technology Data Exchange (ETDEWEB)

    Rodríguez-Romero, Adela, E-mail: adela@unam.mx; Hernández-Santoyo, Alejandra; Fuentes-Silva, Deyanira [Universidad Nacional Autónoma de México, Circuito Exterior, CU, 04310 Coyoacán, DF (Mexico); Palomares, Laura A. [Universidad Nacional Autónoma de México, Apartado Postal 510-3, 62250 Cuernavaca, MOR (Mexico); Muñoz-Cruz, Samira; Yépez-Mulia, Lilian [Centro Médico Nacional Siglo XXI, IMSS, Avenida Cuauhtémoc 330, Colonia Doctores, Mexico, DF (Mexico); Orozco-Martínez, Socorro [Instituto Nacional de Pediatría, Insurgentes Sur 3700C, 04530 Cuicuilco, DF (Mexico); Universidad Nacional Autónoma de México, Circuito Exterior, CU, 04310 Coyoacán, DF (Mexico)

    2014-02-01

    This study describes the three-dimensional structure of the endogenous glycosylated allergen Hev b 2 (endo-β-1,3-glucanase), which exhibits three post-translational modifications that form a patch on the surface of the molecule that is proposed to be an allergenic IgE epitope. Endogenous glycosylated Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis is an important latex allergen that is recognized by IgE antibodies from patients who suffer from latex allergy. The carbohydrate moieties of Hev b 2 constitute a potentially important IgE-binding epitope that could be responsible for its cross-reactivity. Here, the structure of the endogenous isoform II of Hev b 2 that exhibits three post-translational modifications, including an N-terminal pyroglutamate and two glycosylation sites at Asn27 and at Asn314, is reported from two crystal polymorphs. These modifications form a patch on the surface of the molecule that is proposed to be one of the binding sites for IgE. A structure is also proposed for the most important N-glycan present in this protein as determined by digestion with specific enzymes. To analyze the role of the carbohydrate moieties in IgE antibody binding and in human basophil activation, the glycoallergen was enzymatically deglycosylated and evaluated. Time-lapse automated video microscopy of basophils stimulated with glycosylated Hev b 2 revealed basophil activation and degranulation. Immunological studies suggested that carbohydrates on Hev b 2 represent an allergenic IgE epitope. In addition, a dimer was found in each asymmetric unit that may reflect a regulatory mechanism of this plant defence protein.

  10. Targeting Mast Cells and Basophils with Anti-FcεRIα Fab-Conjugated Celastrol-Loaded Micelles Suppresses Allergic Inflammation.

    Science.gov (United States)

    Peng, Xia; Wang, Juan; Li, Xianyang; Lin, Lihui; Xie, Guogang; Cui, Zelin; Li, Jia; Wang, Yuping; Li, Li

    2015-12-01

    Mast cells and basophils are effector cells in the pathophysiology of allergic diseases. Targeted elimination of these cells may be a promising strategy for the treatment of allergic disorders. Our present study aims at targeted delivery of anti-FcεRIα Fab-conjugated celastrol-loaded micelles toward FcεRIα receptors expressed on mast cells and basophils to have enhanced anti-allergic effect. To achieve this aim, we prepared celastrol-loaded (PEO-block-PPO-block-PEO, Pluronic) polymeric nanomicelles using thin-film hydration method. The anti-FcεRIα Fab Fragment was then conjugated to carboxyl groups on drug-loaded micelles via EDC amidation reaction. The anti-FcεRIα Fab-conjugated celastrol-loaded micelles revealed uniform particle size (93.43 ± 12.93 nm) with high loading percentage (21.2 ± 1.5% w/w). The image of micelles showed oval and rod like. The anti-FcεRIα Fab-conjugated micelles demonstrated enhanced cellular uptake and cytotoxity toward target KU812 cells than non-conjugated micelles in vitro. Furthermore, diffusion of the drug into the cells allowed an efficient induction of cell apoptosis. In mouse model of allergic asthma, treatment with anti-FcεRIα Fab-conjugated micelles increased lung accumulation of micelles, and significantly reduced OVA-sIgE, histamine and Th2 cytokines (IL-4, IL-5, TNF-α) levels, eosinophils infiltration and mucus production. In addition, in mouse model of passive cutaneous anaphylaxis, anti-FcεRIα Fab-conjugated celastrol-loaded micelles treatment significantly decreased extravasated evan's in the ear. These results indicate that anti-FcεRIα Fab-conjugated celastrol-loaded micelles can target and selectively kill mast cells and basophils which express FcεRIα, and may be efficient reagents for the treatment of allergic disorders and mast cell related diseases.

  11. Potato lectin activates basophils and mast cells of atopic subjects by its interaction with core chitobiose of cell-bound non-specific immunoglobulin E.

    Science.gov (United States)

    Pramod, S N; Venkatesh, Y P; Mahesh, P A

    2007-06-01

    A major factor in non-allergic food hypersensitivity could be the interaction of dietary lectins with mast cells and basophils. Because immunoglobulin E (IgE) contains 10-12% carbohydrates, lectins can activate and degranulate these cells by cross-linking the glycans of cell-bound IgE. The present objective focuses on the effect of potato lectin (Solanum tuberosum agglutinin; STA) for its ability to release histamine from basophils in vitro and mast cells in vivo from non-atopic and atopic subjects. In this study, subjects were selected randomly based on case history and skin prick test responses with food, pollen and house dust mite extracts. Skin prick test (SPT) was performed with STA at 100 microg/ml concentration. Histamine release was performed using leucocytes from non-atopic and atopic subjects and rat peritoneal exudate cells. SPT on 110 atopic subjects using STA showed 39 subjects positive (35%); however, none showed STA-specific IgE; among 20 non-atopic subjects, none were positive by SPT. Maximal histamine release was found to be 65% in atopic subjects (n = 7) compared to 28% in non-atopic subjects (n = 5); the release was inhibited specifically by oligomers of N-acetylglucosamine and correlates well with serum total IgE levels (R(2) = 0.923). Binding of STA to N-linked glycoproteins (horseradish peroxidase, avidin and IgG) was positive by dot blot and binding assay. As potato lectin activates and degranulates both mast cells and basophils by interacting with the chitobiose core of IgE glycans, higher intake of potato may increase the clinical symptoms as a result of non-allergic food hypersensitivity in atopic subjects.

  12. Test of a theory relating to the cross-linking of IgE antibody on the surface of human basophils

    Energy Technology Data Exchange (ETDEWEB)

    MacGlashan, D.W. Jr.; Dembo, M.; Goldstein, B.

    1985-12-01

    Recent mathematical models of bivalent hapten-induced histamine release from basophils predict that under appropriate conditions histamine release is maximum when cross-link formation is maximum, at a hapten concentration equal to 1/(2K/sub a/), where K/sub a/ is the average affinity constant of the hapten for a single IgE binding site. To test this prediction the authors sensitized human basophils with a monoclonal anti-dinitrophenol IgE and generated histamine release dose-response curves with a bivalent hapten, ..cap alpha..,epsilon-DNP-lysine. The monoclonal IgE has a published affinity constant of 7.1 x 10/sup 7/ M/sup -1/ for epsilon-DNP-lysine as determined by equilibrium dialysis. From the position of the maximum of the histamine dose-response curves, both in the presence and in the absence of monovalent DNP hapten, the authors determine that the sensitizing IgE has an intrinsic affinity constant of 6.9 +/- 0.5 x 10/sup 7/ M/sup -1/ for such that-DNP-lysine and 1.2 +/- 0.6 x 10/sup 6/ M/sup -1/ for ..cap alpha..-DNP-lysine. The agreement between the two estimates of the epsilon-DNP-lysine affinity constant, one from histamine release experiments involving surface bound IgE and one from binding experiments involving IgE free in solution, 1) is consistent with a central prediction of the theory of cross-linking and 2) indicates that the hapten-binding properties of the IgE are unaffected by its being bound to Fc/sub epsilon/ receptors on the basophil surface. 30 references, 3 figures, 3 tables.

  13. Effect of horse gram lectin (Dolichos biflorus agglutinin) on degranulation of mast cells and basophils of atopic subjects: identification as an allergen.

    Science.gov (United States)

    Pramod, Siddanakoppalu N; Krishnakantha, Thirumalai P; Venkatesh, Yeldur P

    2006-11-01

    Horse gram (Dolichos biflorus) is widely consumed in the tropical south Asian countries including rural areas of India. Since D. biflorus agglutinin (DBA) is an important dietary lectin in horse gram, we have studied its effect on the degranulation of mast cells and basophils of atopic subjects. Allergy to horse gram lectin has not been reported so far. Skin prick test (SPT) was performed with 100 microg/mL of DBA. DBA-specific IgE was detected by dot-blot, and ELISA. Histamine release (HR) assay was carried out using leukocytes from non-atopic and atopic subjects, and rat peritoneal exudate cells. Among the atopic group, 10 of 48 subjects (21%) were found to be positive for DBA by SPT, and none were positive in the non-atopic group (n=20). Two subjects out of the ten who tested positive for DBA by SPT were found to be sensitized to DBA as revealed by the presence of specific IgE by ELISA and dot-blot. The HR was found to be 2- to 3-fold higher in DBA-allergic subjects than in non-atopic and atopic subjects. Basophil HR by DBA was found to be similar in both non-atopic and atopic subjects. However, DBA induces activation of mast cells in vivo in a sub-population (21%) of atopic subjects. Two subjects have been identified as having food allergy to horse gram based on the presence of DBA-specific IgE with a positive correlation to basophil HR. This is the first report of food allergy to horse gram, and DBA has been identified as an allergen.

  14. A human monoclonal IgE antibody that binds to MGL_1304, a major allergen in human sweat, without activation of mast cells and basophils.

    Science.gov (United States)

    Ishii, Kaori; Hiragun, Makiko; Hiragun, Takaaki; Kan, Takanobu; Kawaguchi, Tomoko; Yanase, Yuhki; Tanaka, Akio; Takahagi, Shunsuke; Hide, Michihiro

    MGL_1304, a major allergen in human sweat for patients with atopic dermatitis and/or cholinergic urticaria, is secreted from Malassezia globosa on human skin. The amounts of MGL_1304 and IgE against MGL_1304 are evaluated by the histamine release test using basophils or mast cells sensitized with serum containing IgE against MGL_1304, and enzyme linked sorbent assay (ELISA) using MGL_1304 and anti-MGL_1304 antibodies. Here, we identified a human monoclonal IgE (ABS-IgE) that binds to the high affinity IgE receptor (FcεRI) and MGL_1304 with high affinity (KD = 1.99 nM) but does not release histamine from basophils and mast cells. An ELISA using ABS-IgE as a standard IgE revealed that the amount of IgE against MGL_1304 (1000 U/ml) in the standard sera of patients with AD, employed in our previous report, is 32 ng/ml. A sandwich ELISA using ABS-IgE as a detection antibody showed approximately 10 times lower detection limit for MGL_1304 than ELISA in which MGL_1304 is directly bound to an ELISA plate. Moreover, ABS-IgE prevented histamine release from mast cells and basophils by neutralizing MGL_1304 not only in a free form in solution, but also on FcεRI expressed on the cell surface without cell activation. ABS-IgE may be used both to quantify the amount of MGL_1304 and anti-MGL_1304 IgE, and possibly for the treatment of diseases caused/aggravated by type I allergy to MGL_1304.

  15. Histamine release from basophil leukocytes in asthma patients after in vitro provocation with various neuromuscular blocking drugs and intravenous anaesthetic agents

    DEFF Research Database (Denmark)

    Guldager, H; Søndergaard, I

    1987-01-01

    Basophil histamine release is a relatively new investigation technique, which can be used in the diagnosis of anaphylactoid reactions. Our aim in this investigation was to determine reference values for asthma patients and normal subjects. Blood from eight asthmatic patients and eight normal...... subjects was tested for histamine release after in vitro provocation with various neuromuscular blocking drugs and intravenous anaesthetic agents. There was significantly higher histamine release for asthmatic patients than for normal subjects, P less than 0.001 (analysis of variance). This had no effect...

  16. Aeroallergen analyses and their clinical relevance. I. Immunochemical quantification of allergens by RAST-inhibition, Mab-ELISA, basophil histamine release, and counter current immuno electrophoresis

    DEFF Research Database (Denmark)

    Johnsen, C R; Abrahamsen, L; Stahl Skov, P;

    1991-01-01

    The aim was to compare IgE and IgG4 RAST-inhibition assay (RI), monoclonal antibody ELISA (Mab-ELISA), counter current immuno electrophoresis (CCIE) and histamine release from basophil leukocytes (HR) for allergen quantification with special reference to aeroallergen detection. As components...... and Derm. pter. (10-10(2) SQ-U/ml), and 10(2)-10(3) SQ-U/ML for dog. Because of cross-reactivity, a minor degree of interference was observed in the IgE-RI and the HR test for the highest concentration of cat and dog allergens....

  17. The Effects of Electrical Stimuli on Calcium Change and Histamine Release in Rat Basophilic Leukemia Mast Cells

    Science.gov (United States)

    Zhu, Dan; Wu, Zu-Hui; Chen, Ji-Yao; Zhou, Lu-Wei

    2013-06-01

    We apply electric fields at different frequencies of 0.1, 1, 10 and 100 kHz to the rat basophilic leukemia (RBL) mast cells in calcium-containing or calcium-free buffers. The stimuli cause changes of the intracellular calcium ion concentration [Ca2+]i as well as the histamine. The [Ca2+]i increases when the frequency of the external electric field increases from 100 Hz to 10 kHz, and then decreases when the frequency further increases from 10 kHz to 100 kHz, showing a peak at 100 kHz. A similar frequency dependence of the histamine release is also found. The [Ca2+]i and the histamine releases at 100 Hz are about the same as the values of the control group with no electrical stimulation. The ruthenium red (RR), an inhibitor to the TRPV (transient receptor potential (TRP) family V) channels across the cell membrane, is used in the experiment to check whether the electric field stimuli act on the TRPV channels. Under an electric field of 10 kHz, the [Ca2+]i in a calcium-concentration buffer is about 3.5 times as much as that of the control group with no electric stimulation, while the [Ca2+]i in a calcium-free buffer is only about 2.2 times. Similar behavior is also found for the histamine release. RR blockage effect on the [Ca2+]i decrease is statistically significant (~75%) when mast cells in the buffer with calcium are stimulated with a 10 kHz electric field in comparison with the result without the RR treatment. This proves that TRPVs are the channels that calcium ions inflow through from the extracellular environment under electrical stimuli. Under this condition, the histamine is also released following a similar way. We suggest that, as far as an electric stimulation is concerned, an application of ac electric field of 10 kHz is better than other frequencies to open TRPV channels in mast cells, and this would cause a significant calcium influx resulting in a significant histamine release, which could be one of the mechanisms for electric therapy.

  18. Nonreceptor protein tyrosine and lipid phosphatases in type I fc(epsilon) receptor-mediated activation of mast cells and basophils.

    Science.gov (United States)

    Heneberg, Petr; Dráber, Petr

    2002-08-01

    Protein tyrosine and lipid phosphorylations are early and critical events in type 1 Fc(epsilon) receptor (Fc(epsilon)RI)-mediated activation of mast cells and basophils. Tyrosine phosphorylation of Fc(epsilon)RI subunits as well as other signal transduction molecules reflects the balance between the action of protein tyrosine kinases and phosphatases. Similarly, the phosphate content of inositol phospholipids, involved in the recruitment of signalling molecules to the plasma membrane and the generation of secondary messengers, is the net result of the opposing effects of phosphoinositide kinases and lipid phosphatases. This review summarizes the current understanding of the structural and functional aspects of nonreceptor protein tyrosine phosphatases (SHP-1, SHP-2, HePTP, PTP20, PRL1, PRL2, PTP-MEG1 and PTP-MEG2) and lipid phosphatases (SHIP and SHIP2) in the activation of mast cells and basophils after Fc(epsilon)RI aggregation. New approaches towards a deeper understanding of the role of phosphatases in mast cell physiology are also discussed.

  19. Effects of oxatomide and derivatives on high affinity IgE receptor-activated signal transduction pathways in rat basophilic leukemia cells : Role of protein tyrosine hyperphosphorylation and inhibition of extracellular calcium influx

    NARCIS (Netherlands)

    Paulussen, JJC; Fischer, MJE; Roozendaal, RL; van der Heijden, VC; van Dijken, P.; de Mol, NJ; Janssen, LHM

    1998-01-01

    The antiallergic drug oxatomide and analogs inhibit mediator release from a rat basophilic leukemia (RBL-2H3) cell line, which is frequently used as a mast cell model. By investigating a series of derivatives of oxatomide with different inhibiting activities on exocytosis, we aimed to evaluate the r

  20. Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-de pendent upregulation

    Institute of Scientific and Technical Information of China (English)

    Florian HENNERSDORF; Stefan FLORIAN; Andreas JAKOB; Katharina BAUMG(A)RTNER; Karoline SONNECK; Alfred NORDHEIM; Tilo BIEDERMANN; Peter VALENT; Hans-J(o)rg B(U)HRING

    2005-01-01

    Using two-colour flow cytometry >200 antibodies submitted to the 8th International Workshop of Human Leukocyte Differentiation Antigens (HLDA8) have been analyzed for their reactivity with resting and activated CD203c+ basophils.Four antibodies either non-reactive or weakly reactive with resting basophils exhibited an increased reactivity with basophils activated by anti-IgE-mediated cross-linking of the high affinity IgE receptor (FcεRI). These include antibodies against CD164 (WS-80160, clone N6B6 and WS-80162, clone 67D2), as well as two reagents with previously unknown specificities that were identified as CD13 (WS-80274, clone A8) and CD107a (WS-80280, clone E63-880).The activation patterns followed either the "CD203c-like" or "CD63-like" activation profile. The CD203c profile is characterized by a rapid and significant upregulation (of CD13, CD164, and CD203c), reaching maximum levels after 5-15 min of stimulation. The Phosphoinositide-3-kinase (PI3K)-specific inhibitor Wortmannin inhibited the upregulation of these markers whereas 12-O-tetradecanoyl-phorbol-13-acetate (TPA) induced a rapid and FcεRI-independent upregulation within 1-2 min. In the CD63 profile, maximum upregulation (of CD63 and CD107a) was detected only after 20-40 min,and upregulation by TPA reached maximum levels after 60 min. In summary, our data identify CD13, CD107a, and CD164 as novel basophil-activation antigens. Based on time kinetics of upregulation, we hypothesize that molecules of the "CD203c group" and the "CD63 group" are linked to two different mechanisms of basophil activation.

  1. Use of humanised rat basophilic leukaemia cell line RS-ATL8 for the assessment of allergenicity of Schistosoma mansoni proteins.

    Directory of Open Access Journals (Sweden)

    Daniel Wan

    2014-09-01

    Full Text Available Parasite-specific IgE is thought to correlate with protection against Schistosoma mansoni infection or re-infection. Only a few molecular targets of the IgE response in S. mansoni infection have been characterised. A better insight into the basic mechanisms of anti-parasite immunity could be gained from a genome-wide characterisation of such S. mansoni allergens. This would have repercussions on our understanding of allergy and the development of safe and efficacious vaccinations against helminthic parasites.A complete medium- to high-throughput amenable workflow, including important quality controls, is described, which enables the rapid translation of S. mansoni proteins using wheat germ lysate and subsequent assessment of potential allergenicity with a humanised Rat Basophilic Leukemia (RBL reporter cell line. Cell-free translation is completed within 90 minutes, generating sufficient amounts of parasitic protein for rapid screening of allergenicity without any need for purification. Antigenic integrity is demonstrated using Western Blotting. After overnight incubation with infected individuals' serum, the RS-ATL8 reporter cell line is challenged with the complete wheat germ translation mixture and Luciferase activity measured, reporting cellular activation by the suspected allergen. The suitability of this system for characterization of novel S. mansoni allergens is demonstrated using well characterised plant and parasitic allergens such as Par j 2, SmTAL-1 and the IgE binding factor IPSE/alpha-1, expressed in wheat germ lysates and/or E. coli. SmTAL-1, but not SmTAL2 (used as a negative control, was able to activate the basophil reporter cell line.This method offers an accessible way for assessment of potential allergenicity of anti-helminthic vaccine candidates and is suitable for medium- to high-throughput studies using infected individual sera. It is also suitable for the study of the basis of allergenicity of helminthic proteins.

  2. XS-1000i血液分析仪计数嗜碱性粒细胞影响原因分析%An investigation on the accuracy of Sysmex XS-1000i automated hematology analyzer in counting basophils

    Institute of Scientific and Technical Information of China (English)

    邹汉良; 束振华; 梁汉章; 张松; 陈丕绩

    2011-01-01

    目的 分析Sysmex XS- 1000i血液分析仪计数嗜碱性粒细胞部分标本出现假性增高的影响原因.方法 对50例外周血中性粒细胞形态正常的标本使用Sysmex XE-5000、XS-1000i血液分析仪进行嗜碱性粒细胞计数,作相关统计学分析.4例使用Sysmex XS- 1000i血液分析仪计数嗜碱性粒细胞出现明显增高标本与手工计数法进行比较,做相关形态学分析.结果 50例外周血中性粒细胞形态正常的标本使用Sysmex XE-5000、XS-1000i血液分析仪进行嗜碱性粒细胞计数,两者相关系数r=0.715,嗜碱性粒细胞计数差异具统计学意义(P<0.01).4例使用Sysmex XS-1000i血液分析仪计数嗜碱性粒细胞出现明显增高标本均出现中性粒细胞中毒颗粒、退行性变等形态学改变.结论 两仪器间计数嗜碱性粒细胞结果差异有统计学意义,外周血中性粒细胞中毒颗粒等形态改变会造成XS-1000i血液分析仪计数嗜碱性粒细胞假性增高,对嗜碱性粒细胞异常增高的结果需进行手工计数分类.%Objective To identify the cause underlying the false increase in basophil count tested by Sysmex XS-1000i automated hematology analyzer. Methods 50 peripheral blood samples with normal neutrophil morphology were compared between Sysmex XE-5000 and XS-1000i analyzers for the count of basophils. 4 blood samples with obvious increase in the basophil count tested by Sysmex XS-1000i automated hematology analyzer, were compared with the results from manual analysis. Results The correlation test about basophil ratio, with a correlation index (r) of 0.715. There were obvious differences in basophil ratios (P<0.01). Toxic granulation and degradation were detected manually in neutrophils in all 4 samples with falsely increased basophils analyzed by Sysmex XS-1000i. Conclusions There was difference between two machines in counting basophil. We demonstrate that toxic granulation and degradation in neutrophils of peripheral blood may

  3. Expressions of CD63 and CD203c in basophils of allergic asthma patients%过敏性哮喘患者嗜碱粒细胞CD63和CD203c的表达

    Institute of Scientific and Technical Information of China (English)

    杨玲; 李强; 王树云; 许以平; 郭胤仕; 姚迪; 史桂英

    2012-01-01

    目的:探讨过敏性哮喘患者嗜碱粒细胞激活后其CD63和CD203c的表达情况.方法:采用流式细胞术检测过敏性哮喘(哮喘组)、非过敏性哮喘患者(非过敏性哮喘组)及正常对照者(正常对照组)的血嗜碱性粒细胞中CD63和CD203c的表达,并进行比较.结果:以CD123和人类白细胞抗原DR联合设门,可观察到外周血中的嗜碱性粒细胞.与非过敏性哮喘组及正常对照组比较,过敏性哮喘组患者的血嗜碱粒细胞CD63荧光强度显著增加(P<0.01).经粉尘螨刺激后,与刺激前对照、非过敏性哮喘者及正常对照者比较,过敏性哮喘患者的血嗜碱粒细胞CD203c和CD63荧光强度均显著增加(P<0.01).嗜碱粒细胞CD203c、CD63表达程度与皮试结果及特异性IgE水平间均存在不同程度的相关性.受试者工作特征曲线分析结果显示,嗜碱粒细胞活化试验的曲线下面积为0.923,高于特异性IgE的曲线下面积(0.876).结论:采用流式细胞术检测CD63表达来确定活化嗜碱粒细胞是诊断过敏性疾病的一种安全、有效的体外检测方法.%Objective To study the expressions of CD63 and CD203c in activated basophils of patients with allergic asthma. Methods Flow cytometry (FCM) was used to measure the expressions of CD63 and CD203c in blood basophils of allergic asthma patients, non-allergic asthma patients and normal controls, and the results were compared. Results Peripheral blood basophils could be selected by using CD 123 and HLA-DR as the gate. The median fluorescence intensity (MFI) of CD63 in basophils of allergic asthma patients increased markedly when compared with that of non-allergic asthma patients and normal controls (P<0.01). After stimulated with dust mites, the median fluorescence intensity of CD203c and CD63 in basophils of allergic asthma patients increased significantly when compared with that before stimulation and that of non-allergic asthma patients and normal controls (P<0.01). The

  4. Controlled-release formulation of antihistamine based on cetirizine zinc-layered hydroxide nanocomposites and its effect on histamine release from basophilic leukemia (RBL-2H3 cells

    Directory of Open Access Journals (Sweden)

    Hussein Al Ali SH

    2012-07-01

    Full Text Available Samer Hasan Hussein Al Ali,1 Mothanna Al-Qubaisi,2 Mohd Zobir Hussein,1,3 Maznah Ismail,2,4 Zulkarnain Zainal,1 Muhammad Nazrul Hakim51Department of Chemistry, Faculty of Science, 2Laboratory of Molecular Biomedicine, Institute of Bioscience, 3Advanced Materials and Nanotechnology Laboratory, Institute of Advanced Technology (ITMA, 4Department of Nutrition and Dietetics, Faculty of Medicine and Health Science, Universiti Putra, Malaysia; 5Department of Biomedical Science, Faculty of Medicine and Health Science, Universiti Putra Malaysia, Selangor, MalaysiaAbstract: A controlled-release formulation of an antihistamine, cetirizine, was synthesized using zinc-layered hydroxide as the host and cetirizine as the guest. The resulting well-ordered nanolayered structure, a cetirizine nanocomposite "CETN," had a basal spacing of 33.9 Å, averaged from six harmonics observed from X-ray diffraction. The guest, cetirizine, was arranged in a horizontal bilayer between the zinc-layered hydroxide (ZLH inorganic interlayers. Fourier transform infrared spectroscopy studies indicated that the intercalation takes place without major change in the structure of the guest and that the thermal stability of the guest in the nanocomposites is markedly enhanced. The loading of the guest in the nanocomposites was estimated to be about 49.4% (w/w. The release study showed that about 96% of the guest could be released in 80 hours by phosphate buffer solution at pH 7.4 compared with about 97% in 73 hours at pH 4.8. It was found that release was governed by pseudo-second order kinetics. Release of histamine from rat basophilic leukemia cells was found to be more sensitive to the intercalated cetirizine in the CETN compared with its free counterpart, with inhibition of 56% and 29%, respectively, at 62.5 ng/mL. The cytotoxicity assay toward Chang liver cells line show the IC50 for CETN and ZLH are 617 and 670 µg/mL, respectively.Keywords: cetirizine hydrochloric acid

  5. 嗜盐嗜碱放线菌的研究进展与展望%Research on Progress of Halophilic Basophilic Actinomycetes

    Institute of Scientific and Technical Information of China (English)

    加苏尔·阿不都克里木; 旭格拉; 塔衣尔; 阿不都克里木·热依木; 木尼热木·阿力木江; 迪丽拜尔·托合提

    2015-01-01

    Haloalkaliphilic extremophiles were survied in saline alkali soil. Halophilic basophilic Actinomycetes with the genetic characteristics and the special supersedes way, produced a variety of secondary me⁃tabolites, was one of microbial resources with great application prospect. This paper mainly reviewed Halophilic ba⁃sophilic Actinomycetes Resources, standard classification, distribution, system science, development and utilization prospects.%嗜盐嗜碱极端微生物主要分布于地球上的盐碱土壤中,嗜盐嗜碱放线菌适应并长期生存这一特殊极端环境,已具备独特的遗传特性和新陈代谢途径,能够产生具有开发利用价值的多种次生代谢产物,是一类极具应用前景的微生物资源。文章主要综述了嗜盐嗜碱放线菌的资源、分类标准、分布、系统学及其发展趋势、开发利用前景。

  6. Analysis of allergens in tubeimu saponin extracts by using rat basophilic leukemia 2H3 cell-based affinity chromatography coupled to liquid chromatography and mass spectrometry.

    Science.gov (United States)

    Zhang, Tao; Han, Shengli; Liu, Qi; Guo, Ying; He, Langchong

    2014-11-01

    An affinity two-dimensional chromatography method was developed for the recognition, separation, and identification of allergic components from tubeimu saponin extracts, a preparation often injected to treat various conditions as indicated by traditional Chinese medicine. Rat basophilic leukemia-2H3 cell membranes were used as the stationary phase of a membrane affinity chromatography column to capture components with affinity for mast cells that could be involved in a degranulation reaction. The retained components were enriched and analyzed by membrane affinity chromatography with liquid chromatography and mass spectrometry via a port switch valve. Suitability and reliability of the method was investigated using appropriate standards, and then, the method was applied to identify components retained from tubeimu saponin extracts. Tubeimoside A was identified in this way as a potential allergen, and degranulation assays confirmed that tubeimoside A induces RBL-2H3 cell degranulation in a dose-dependent manner. An increase in Ca(2+) influx indicated that degranulation induced by tubeimoside A is likely Ca(2+) dependent. Coupled with the degranulation assay, RBL-2H3 cell-based affinity chromatography coupled with liquid chromatography and mass spectrometry is an effective method for screening and identifying allergic components from tubeimu saponin extracts.

  7. Chorea as a clinical feature of the basophilic inclusion body disease subtype of fused-in-sarcoma-associated frontotemporal lobar degeneration.

    Science.gov (United States)

    Kawakami, Ito; Kobayashi, Zen; Arai, Tetsuaki; Yokota, Osamu; Nonaka, Takashi; Aoki, Naoya; Niizato, Kazuhiro; Oshima, Kenichi; Higashi, Shinji; Katsuse, Omi; Hosokawa, Masato; Hasegawa, Masato; Akiyama, Haruhiko

    2016-04-04

    Choreoathetoid involuntary movements are rarely reported in patients with frontotemporal lobar degeneration (FTLD), suggesting their exclusion as a supportive feature in clinical diagnostic criteria for FTLD. Here, we identified three cases of the behavioral variant of frontotemporal dementia (bvFTD) that display chorea with fused in sarcoma (FUS)-positive inclusions (FTLD-FUS) and the basophilic inclusion body disease (BIBD) subtype. We determined the behavioral and cognitive features in this group that were distinct from other FTLD-FUS cases. We also reviewed the clinical records of 72 FTLD cases, and clarified additional clinical features that are predictive of the BIBD pathology. Symptom onset in the three patients with chorea was at 44.0 years of age (±12.0 years), and occurred in the absence of a family history of dementia. The cases were consistent with a clinical form of FTD known as bvFTD, as well as reduced neurological muscle tone in addition to chorea. The three patients showed no or mild parkinsonism, which by contrast, increased substantially in the other FTLD cases until a later stage of disease. The three patients exhibited severe caudate atrophy, which has previously been reported as a histological feature distinguishing FTLD-FUS from FTLD-tau or FTLD-TAR DNA-binding protein 43. Thus, our findings suggest that the clinical feature of choreoathetosis in bvFTD might be associated with FTLD-FUS, and in particular, with the BIBD subtype.

  8. OM-X®, Fermented Vegetables Extract Suppresses Antigen-Stimulated Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells and Passive Cutaneous Anaphylaxis Reaction in Mice.

    Science.gov (United States)

    Itoh, Tomohiro; Miyake, Yasuyoshi; Kasashima, Takuya; Shimomiya, Yoshie; Nakamura, Yuki; Ando, Masashi; Tsukamasa, Yasuyuki; Takahata, Muneaki

    2015-09-01

    OM-X® is a hand-made and naturally manufactured probiotic supplement. This fermented food product is made from vegetables, fruits, seaweeds and mushrooms, using 12 strains of lactic acid bacteria and bifidobacteria. OM-X® is also known to have beneficial health properties, and some of its components show effects on antigen (Ag)-stimulated degranulation activity, indicating that OM-X® may be useful in the treatment of allergy responses and symptoms. In this study, we evaluated the inhibitory effects of OM-X® on Ag-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells, clarified the underlying mechanisms, and determined the active compounds in OM-X® for suppression of degranulation. Treatment with OM-X® gradually suppressed Ag-stimulated degranulation throughout the maturation period. OM-X® also gradually produced melanoidins by lactic acid bacterial fermentation during the maturation process. There was a high correlation between the suppression levels of Ag-stimulated degranulation and the browning of OM-X®. Furthermore, the inhibition of Ag-stimulated degranulation by OM-X® was found to be partially due to the direct inactivation of NADPH oxidase. To elucidate the in vivo effects of OM- X®, type I allergy model mice were orally administered with OM-X®, and the passive cutaneous anaphylaxis (PCA) reaction was measured. OM-X® intake remarkably suppressed the PCA reaction. Taken together, our findings suggest that OMX® could be a beneficial food to ameliorate allergic reactions.

  9. 微波处理对嗜碱和嗜盐海洋放线菌分离效果的影响%Effects of microwave irradiation on isolation of basophilic and halophilic marine actinomycetes

    Institute of Scientific and Technical Information of China (English)

    丁彦博; 蔡超靖; 穆云龙; 单越琦; 路新华; 蒋沁

    2012-01-01

    [Objective] Study the effects of microwave irradiation on isolation of basophilic and halophilic marine actinomycetes. [Methods] Seven marinemud samples were radiated by microwave and then gradient diluted for isolation of basophilic and halophilic marine actinomycetes in three media. [Results] Microwave irradiation could highly significantly increase the total quantity of basophilic and halophilic marine actinomycetes respectively in four and three marinemud samples. The total quantity of basophilic and halophilic marine rare actinomycetes of Micromonospora, Actinoplanes and Nocardia were significantly increased after microwave irradiation. The species of other marine rare actinomycetes such as Catellatospora, Microbispora, Streptosporangium were increased to one to four in different samples. [Conclusion] Microwave irradiation could significantly increase the total quantity of basophilic and halophilic marine actinomycetes and the species of marine rare actinomycetes.%[目的]研究微波处理对于分离嗜碱和嗜盐海洋放线菌的效果.[方法]用微波处理7份海泥样品,梯度稀释后涂布于3种分离培养基,分离具有嗜碱和嗜盐特性的海洋放线菌.[结果]微波处理后的7份样品中,4份样品中嗜碱海洋稀有放线菌和3份样品的嗜盐海洋稀有放线菌数量极显著提高;7份样品中的嗜碱、嗜盐海洋小单孢菌属、游动放线菌属、诺卡氏菌属等稀有放线菌数量均有显著增加,不同样品中新分离到链孢菌属、小双孢菌属、链孢囊菌属及其他未鉴定的海洋稀有放线菌,分离到属的数量提高了1-4个.[结论]微波处理不仅显著提高嗜碱和嗜盐海洋放线菌的分离数量,而且明显增加了海洋稀有放线菌的分离种类.

  10. The clinical values of peripheral blood basophil activation test in children with allergic diseases%过敏性疾病患儿外周血嗜碱性粒细胞百分比及活化率检测

    Institute of Scientific and Technical Information of China (English)

    杨皓征; 曹兰芳; 王利民

    2011-01-01

    Objective To explore the clinical values of basophil activation test in childhood allergic diseases by the analysis of CRTH2+ cells, percent of basophil, and activation of basophil in peripheral blood.Methods The peripheral basophils activation in healthy children and allergic patients ( asthma and/or rhinitis and/or dermatitis ) were analyzed with a combination of CD3/CRTH2/CD63 antibodies by flow cytometric.Results Ninty-three subjects ( 25 in control group, 68 in allergy group ) completed the study.The peripheral CRTH2+ cell level in the allergy group ( 0.53% ± 0.22% ) was significantly higher than that in control group ( 0.44% ± 0.14% )( P < 0.05 ).The peripheral basophil level of the allergy group ( 0.39% ± 0.19% ) was significantly higher than that of control group ( 0.130% ± 0.073% )( P < 0.01 ).The basophil activation ratio was higher in allergic patients ( 85.77% ± 10.63% ) than that in control group ( 31.62% ± 17.41% ).In addition, Spearman correlation coefficients indicated a moderate positive correlation between basophil activation ratio and slgE calss categorized by Unicap calss.Moreover, Spearman correlation coefficients indicated medium positive correlation between basophil activation ratio and standardized skin prick test.Conclusions The peripheral CRTH2+ cell level and basophil level increase in allergic children.It suggests that basophils play an important role in the pathogenesis of allergic diseases.Moreover, basophil activation test may have some values in the diagnosis and screening of allergic diseases in children.%目的 探讨嗜碱性粒细胞在儿童过敏性疾病中的临床参考价值.方法 应用流式细胞仪,采用识别CRTH2/CD63/CD4组合抗体,检测过敏性疾病患儿与正常儿童外周血中CRTH2阳性细胞和嗜碱性粒细胞百分比及其活化率,并将检测结果与血清总IgE、特异性IgE、皮肤点刺试验结果作比较,分析其内在联系.结果 68例过

  11. 2株嗜碱性放线菌的分离鉴定及特性研究%Isolation, identification and characterization of two basophilic actinomycosis

    Institute of Scientific and Technical Information of China (English)

    熊鹰; 余金辉; 阿不都克里木·热依木; 魏艳红; 李逸; 莫小妍; 陈湖星; 袁永泽; 刘德立

    2012-01-01

    从新疆巴洲和硕县碱性土壤中分离纯化出2株放线菌菌株XJ-1和XJ-4,对其形态特征、生理生化特性、抗生素产生以及16SrRNA基因序列分析等方面进行了多种特性研究.结果表明.2株放线菌在pH7.0的中性条件下不能生长,在pH12.0的强碱性条件下能够生长,最适生长pH为10.0,属于嗜碱性放线菌.2株放线菌菌株能耐受10%的NaCl,最高耐受温度为45℃.菌株XJ-1产生抗生素,能抑制柑橘绿霉菌、水稻轮纹菌、棉花枯萎菌和小麦赤霉菌的生长.16S rRNA基因同源性分析表明XJ-1属于拟诺卡氏菌属的Nocardiopsis dassonvillei,序列相似性达到了99.9%.而XJ-4与Nocardiopsis.sp.AF-333的序列相似性达到了99.3%,有可能是一新种.%Two actinomycete strains XJ-1 and XJ-4 were isolated from the alkaline soil of Shuo County and BaZhou County in Xinjiang Province. Based on the morphology, physiological and biochemical characteristics, antibiotics production and 16S rRNA gene sequence analysis, the results showed that the optimum growth pH of these two baso-philic actinomycosis was 10. 0; they could not grow in the neutral conditions of pH7. 0 and could grow in the strong alkaline conditions of pH12. 0. Two actinomycete strains had a tolerance of 10% NaCl and the highest tolerance of temperature were 45℃. XJ-1 was able to produce antibiotics. It could obviously inhibit the growth of Penicillium digi-tatum, but also inhibit the growth of Rhizoctonia solani, Fusariumoxysporum and Fu-sarium raminearum. XJ-1 was preliminarily determined to be the Nocardiopsis dassonvil-lei, their sequence similarity reached 99. 9%. The homology of the 16S rDNA sequences of strain XJ-4 with Nocardiopsis. sp. AF-333 was more than 99. 0% , so the strain XJ-4 maybe a new species belonged to the basophilic actinomycetes.

  12. JNK-dependent NFATc1 pathway positively regulates IL-13 gene expression induced by (-)-epigallocatechin-3-gallate in human basophilic KU812 cells.

    Science.gov (United States)

    Wu, Haitao; Qi, Hang; Iwasaki, Dai; Zhu, Beiwei; Shimoishi, Yasuaki; Murata, Yoshiyuki; Nakamura, Yoshimasa

    2009-10-01

    (-)-Epigallocatechin-3-gallate (EGCG) has been reported to possess a wide range of biological and pharmacological properties. In this study, we investigated the effects of EGCG on IL-13 gene expression in human basophilic KU812 cells. The IL-13 mRNA expression level was dose-dependently increased by treatment with EGCG (5-20 microM) for 1 h and additional incubation in a medium for 23 h. EGCG significantly increased the intracellular peroxide level as detected by the peroxide-sensitive probe 2',7'-dichlorodihydrofluorescein diacetate. A pharmacological experiment using catalase and a structure-activity relationship study revealed that the exogenously produced H(2)O(2) significantly, but partially, contributed to the IL-13 expression as well as the intracellular oxidative status. Furthermore, EGCG at the concentration required for IL-13 up-regulation activated c-Jun NH(2)-terminal kinase (JNK), but not extracellular signal-regulated protein kinase or p38 mitogen-activated protein kinase in KU812 cells. Transfection of a JNK-specific siRNA as well as treatment with a JNK-specific inhibitor, SP600125, significantly reduced the EGCG-induced IL-13 mRNA expression, by 47.1 and 44.6%, respectively. In addition, we observed the nuclear translocation, mRNA up-regulation, and activation of DNA binding with the IL-13 promoter of nuclear factor of activated T cells (NFATc1) in the EGCG-treated cells. These data provide biological evidence that EGCG induces IL-13 mRNA expression via the JNK-dependent NFATc1 pathway in KU812 cells.

  13. Aeroallergen analyses and their clinical relevance. I. Immunochemical quantification of allergens by RAST-inhibition, Mab-ELISA, basophil histamine release, and counter current immuno electrophoresis.

    Science.gov (United States)

    Johnsen, C R; Abrahamsen, L; Stahl Skov, P; Johansen, N; Poulsen, L K

    1991-10-01

    The aim was to compare IgE and IgG4 RAST-inhibition assay (RI), monoclonal antibody ELISA (Mab-ELISA), counter current immuno electrophoresis (CCIE) and histamine release from basophil leukocytes (HR) for allergen quantification with special reference to aeroallergen detection. As components of indoor aeroallergens, cat, dog, and Derm. pter. allergen extracts were selected for the experiments. To evaluate unspecific interference, these allergens were compared mutually and with Cladosporium herbarum. Allergen extracts in varying dilutions were mixed with crushed glass fibre filter materials, eluted, recovered by centrifugation, and allergen concentration quantified by the assays. Equal sensitivity was found for both IgE- and IgG4-RI assaying cat allergen (in the range 5-50 SQ-U/ml) and dog allergen (in the range 10(2)-10(3) SQ-U/ml). The IgG4-RI assaying Derm. pter. was more sensitive (50 SQ-U/ml) than IgE-RI (2*10(3) SQ-U/ml). The ranges of allergen detection limits for the Mab-ELISA were equal for cat and Derm. pter. (10-10(2) SQ-U/ml). The range of allergen detection limits for CCIE, assaying dog were 10(4)-10(5) SQ-U/ml. The ranges of allergen detection limits for HR were equal for cat and Derm. pter. (10-10(2) SQ-U/ml), and 10(2)-10(3) SQ-U/ML for dog. Because of cross-reactivity, a minor degree of interference was observed in the IgE-RI and the HR test for the highest concentration of cat and dog allergens.

  14. 活化嗜碱粒细胞在变应性鼻炎患者外周血中的表达%Expression of activated basophile in peripheral blood in patients with allergic rhinitis

    Institute of Scientific and Technical Information of China (English)

    张淳; 石嘉俪; 徐雅男; 王利民; 王家东

    2011-01-01

    目的 探讨嗜碱粒细胞在变应性鼻炎患者外周血中数量及活化率的变化及其临床意义.方法 以化学诱导趋向性受体单抗作为特异性的荧光标记,采用三色免疫荧光标记流式细胞术对20例急性发作期变应性鼻炎患者(急性组)、20例反复发作变应性鼻炎患者(慢性组)及30例健康者(对照组)的外周血嗜碱粒细胞及其活化率进行定量检测.结果 慢性组外周血化学诱导趋向性受体及嗜碱粒细胞高于急性组(P<0.01)和对照组(P<0.05),对照组高于急性组(P<0.01).慢性组与急性组嗜碱粒细胞活化率均高于对照组(P<0.01).结论 嗜碱粒细胞活化是导致变应性鼻炎症状出现的主要因素之一.慢性期患者嗜碱粒细胞的持续增高和活化可能与变应性鼻炎反复发作有关.%Objective To evaluate the number and the activation rate of basophile in peripheral blood in patients with allergic rhinitis and their clinical significance. Methods The chemoattractant receptor-homologus molecule expressed on T helper cells was used as a specific fluorescent marker. Twenty patients with acute allergic rhinitis, 20 patients with chronic allergic rhinitis and 30 healthy controls were evaluated the number and the activation rate of basophile in peripheral blood with three-color fluorescent flow cytometry analysis. Results The numbers of chemoattractant receptor-homologus molecule expressed on T helper cells and basophile were significantly larger in chronic allergic rhinitis group than those in acute allergic rhinitis group (P<0. 01) and normal group (P< 0. 05). The activation rate of basophile was significantly higher in acute allergic rhinitis and chronic allergic rhinitis group than that in control group(P<0. 01). Conclusion The activation of basophile is one of the main factors inducing the symptoms of allergic rhinitis. The continuous increase of the number and the activation rate of basophile may be involved in

  15. Direct interaction of Syk and Lyn protein tyrosine kinases in rat basophilic leukemia cells activated via type I Fc epsilon receptors.

    Science.gov (United States)

    Amoui, M; Dráberová, L; Tolar, P; Dráber, P

    1997-01-01

    Activation of rat mast cells through the receptor with high affinity for IgE (Fc epsilonRI) requires a complex set of interactions involving transmembrane subunits of the Fc epsilonRI and two classes of nonreceptor protein tyrosine kinase (PTK). the Src family PTK p53/p56(lyn) (Lyn) and the Syk/ZAP-family PTK p72(syk) (Syk). Early activation events involve increased activity of Lyn and Syk kinases and their translocation into membrane domains containing aggregated Fc epsilonRI, but the molecular mechanisms responsible for these changes have remained largely unclear. To determine the role of Fc epsilonRI subunits in this process, we have analyzed Syk- and Lyn-associated proteins in activated rat basophilic leukemia (RBL) cells and their variants deficient in the expression of Fc epsilonRI beta or gamma subunits. Sepharose 4B gel chromatography of postnuclear supernatants from Nonidet-P40-solubilized antigen (Ag)- or pervanadate-activated RBL cells revealed extensive changes in the size of complexes formed by Lyn and Syk kinases and other cellular components. A fusion protein containing Src homology 2 (SH2) and SH3 domains of Lyn bound Syk from lysates of nonactivated RBL cells; an increased binding was observed when lysates from Ag- or pervanadate-activated cells were used. A similar amount of Syk was bound when lysates from pervanadate-activated variant cells deficient in the expression of Fc epsilonRI beta or gamma subunits were used, suggesting that Fc epsilonRI does not function as the only intermediate in the formation of the Syk-Lyn complexes. Further experiments have indicated that Syk-Lyn interactions occur in Ag-activated RBL cells under in vivo conditions and that these interactions could involve direct binding of the Lyn SH2 domain with phosphorylated tyrosine of Syk. The physical association of Lyn and Syk during mast-like cell activation supports the recently proposed functional cooperation of these two tyrosine kinases in Fc epsilonRI signaling.

  16. Use of humanized rat basophilic leukemia reporter cell lines as a diagnostic tool for detection of allergen-specific IgE in allergic patients: time for a reappraisal?

    Science.gov (United States)

    Falcone, Franco H; Alcocer, Marcos J C; Okamoto-Uchida, Yoshimi; Nakamura, Ryosuke

    2015-11-01

    The interaction between allergens and specific IgE is at the heart of the allergic response and as such lies at the center of techniques used for diagnosis of allergic sensitization. Although serological tests are available, in vivo tests such as double-blind placebo-controlled food challenges (DBPCFC) and skin prick test (SPT) associated to the patients' clinical history are still the main guides to clinicians in many practices around the world. More recently, complex protein arrays and basophil activation tests, requiring only small amounts of whole blood, have been developed and refined, but are yet to enter clinical practice. Similarly, the use of rat basophilic leukemia (RBL) cell lines for detection of allergen-specific IgE has been made possible by stable transfection of the human FcεRI α chain into this cell line more than 20 years ago, but has not found widespread acceptance among clinicians. Here, we review the perceived limitations of diagnostic applications of humanized RBL systems. Furthermore, we illustrate how the introduction of reporter genes into humanized RBL cells is able to overcome most of these limitations, and has the potential to become a new powerful tool to complement the armamentarium of allergists. A demonstration of the usefulness of humanized RBL reporter systems for elucidation of complex IgE sensitization patterns against wheat proteins and a section on the use of fluorescence-based reporter systems in combination with allergen arrays close the review.

  17. Urticaria crónica: Evolución clínica, prueba del suero autologo, recuento y activación de basofilos Chronic urticaria: Follow up, autologous serum skin test, basophil count and activation

    Directory of Open Access Journals (Sweden)

    N. Galassi

    2003-01-01

    Full Text Available Recientes estudios sugieren que algunos pacientes con urticaria crónica tienen autoanticuerpos dirigidos contra la IgE o a la cadena alfa de su receptor de alta afinidad FceRIa. La detección de éstos se realiza in vivo mediante la prueba del suero autólogo (PSA o in vitro mediante una variedad de técnicas. Describimos aquí nuestra experiencia con 37 pacientes, 28 mujeres y 9 varones con una relación femenino/masculino de 3.1 y edad promedio de 36.5 años (16-78. La PSA fue positiva en 25 (68% y negativa en 12 (32%. El suero indujo una pápula significativamente más grande que el plasma (122 ± 78 mm² vs 57 ± 66 mm², pRecent advances on the pathogenesis of chronic urticaria have defined a group of patients with autoantibodies directed to the IgE or to the alpha chain of the Fc high affinity receptor of IgE, FceRIa. These antibodies are detected in vivo through the autologous serum test (AST and in vitro with a variety of techniques. We here describe 37 patients with chronic urticaria, 28 female and 9 male, with a f/m ratio of 3.1. Mean age at onset was 36.5 years (range 16-78. AST was positive in 25 (68% of 37 patients. Serum induced a wheal significantly larger than plasma (122 ± 78 mm² vs 57 ± 66 mm², p<0.05. Median persistence of the chronic urticaria, estimated by Kaplan-Meyer analysis, was 437 days, with no difference between AST(+ and AST(- patients (437 vs. 369, p=0.18. Mean IgE concentration was 157 ± 173 IU/mL, as expected in an unselected population. Basophil count was lower in patients compared with controls (17 ± 12 cel/mL vs. 43 ± 27 cel/mL, p< 0.008. Only sera from 2/7(28.6% patients AST (+ and very low basophil count consistently induced expression of CD63. This effect was abrogated in non-releasing basophils, confirming the presence of antibodies directed to the FceRIa-IgE . We conclude that functional antibodies are present in only a minority of patients and that their identification does not predict the outcome.

  18. 嗜碱粒细胞组胺释放试验检测抗高亲和力IgE抗体及其受体%Significance of basophil histamine release assays in chronic urticaria

    Institute of Scientific and Technical Information of China (English)

    刘荣卿; 孙仁山

    2001-01-01

    Objective To Investigate the pathogenesis of chronic idiopathic urticaria.Methods Basophil histamine release assay was utilized.Result 15 of 32 cases(46.9%)had increased serum histamine releasing activities. This implied that anti-FcεR1 and anti-IgE autoantibodies existed in the sera of some patients with chronic idiopathic urticaria.Conclusion It suggested that autoimmunity might take part in the occurrence of some patients with chronic idiopathic urticaria.%目的 探讨慢性荨麻疹的发生机制。方法 用嗜碱粒细胞组胺释放试验,检测慢性特发性荨麻疹患者的血清组胺释放活性。结果 32例中,有15例(46.9%)患者血清组胺释放活性增高,提示抗FcεR1或抗IgE自身抗体的存在。结论 部分慢性荨麻疹的发生与自身免疫机制有关。

  19. Gambaran Sel Eosinofil, Monosit, dan Basofil Setelah Pemberian Spirulina pada Ayam yang Diinfeksi Virus Flu Burung (OBSERVATION OF EOSINOPHILS, MONOCYTES, AND BASOPHILS AFTER TREATED WITH SPIRULINA IN CHICKENS THAT INFECTED WITH AVIAN INFLUENZA VIRUS

    Directory of Open Access Journals (Sweden)

    Widya Paramita Lokapirnasari

    2015-05-01

    Full Text Available High Pathogenecity Avian Influenza (HPAI viruses have high virulence and can frequently causesudden death on birds. The aims of this research was to know the role of Spirulina to a number ofmonocytes and lymphocytes in the blood of chickens which infected with the H5N1 virus. This researchconsisted of three levels of treatment in which each level given Spirulina 0%, 10%, 20% in the fresh wateralgae as drinking water. Each treatment consisted of seven replicates, and the treatment was done sincethe chickens at age 19 until 44 days ( for 25 days. Artificial infection of the chickens with the virus waschallenged by using AI (H5N1 104 EID 50 (A/Ck/Indonesia/BL/03 with route to the respiratory tract (nosedrops 0,1 mL starting on day 19. The results showed that there were a significant difference (p<0.05 ontreatment that given Spirulina at doses of 0%, 10% and 20% for the number ofn monocytes, eosinophils,whereas no significant difference (p > 0.05 was observed in basophils.

  20. Serotonin storage pools in basophil leukemia and mast cells: characterization of two types of serotonin binding protein and radioautographic analysis of the intracellular distribution of (/sup 3/H)serotonin

    Energy Technology Data Exchange (ETDEWEB)

    Tamir, H. (New York Psychiatric Inst., New York); Theoharides, T.C.; Gershon, M.D.; Askenase, P.W.

    1982-06-01

    The binding of serotonin to protein(s) derived from rat basophil leukemia (RBL) cells and mast cells was studied. Two types of serotonin binding protein in RBL cells was found. These proteins differed from one another in molecular weight and eluted in separate peaks from sephadex G-200 columns. Peak I protein (KD = 1.9 x 10/sup -6/ M) was a glycoprotein that bound to concanavalin A (Con A); Peak II protein (KD/sub 1/ = 4.5 x 10/sup -/8 M; KD/sub 2/ = 3.9 x 10/sup -6/ M) did not bind to Con A. Moreover, binding of (/sup 3/H)serotonin to protein of Peak I was sensitive to inhibition by reserpine, while binding of (/sup 3/H)serotonin to protein of Peak II resisted inhibition by that drug. Other differences between the two types of binding protein were found, the most significant of which was the far more vigorous conditions of homogenization required to extract Peak I than Peak II protein. Electron microscope radioautographic analysis of the intracellular distribution of (/sup 3/H) serotonin taken up in vitro by RBL cells or in vivo by murine mast cells indicated that essentially all of the labeled amine was located in cytoplasmic granules.No evidence for a pool in the cytosol was found and all granules were capable of becoming labeled. The presence of two types of intracellular serotonin binding proteins in these cells may indicate that there are two intracellular storage compartments for the amine. Both may be intragranular, but Peak I protein may be associated with the granular membrane while Peak II protein may be more free within the granular core. Different storage proteins may help to explain the differential release of amines from mast cell granules.

  1. Rabbit IgG directed to a synthetic C-terminal peptide of the major grass pollen allergen Lol p I inhibits human basophil histamine release induced by natural Lol p I.

    Science.gov (United States)

    van Ree, R; Aalberse, R C

    1995-03-01

    The potential role of allergen-specific IgG antibodies as 'blocking' antibodies in allergen-induced human basophil histamine release was investigated. This was studied in a model with the major grass pollen allergen Lol p I and polyclonal rabbit antisera directed against this allergen and against a synthetic peptide of its C terminus. When allergen and antibodies were allowed to preincubate, Lol p I induced histamine release was inhibited up to 85% by the antiserum against Lol p I. By omitting preincubation, and thereby more closely mimicking an in vivo situation, up to 55% inhibition was realized. This indicates that allergen-specific IgG can act as 'blocking' antibody without preincubation. Immunization of rabbits with a synthetic C-terminal peptide of Lol p I resulted in antibodies reactive with natural Lol p I. Despite their 100-fold lower avidity for Lol p I (as compared with antinatural Lol p I), these antibodies had the capacity to inhibit Lol p I induced histamine release for > 90% (up to 50% without preincubation). This indicates that it is possible to block histamine release induced by a major allergen with low-avidity IgG antibodies directed against a minor proportion of the allergen (25 amino acids). IgE antibodies from the donors studied were unreactive with this synthetic peptide, indicating that for blocking activity identical epitope specificity of IgE and IgG is not essential. This opens interesting perspectives for application of synthetic peptides in immunotherapy, distinct from their effects on T cell reactivity.

  2. The effect of specific immunotherapy on NK cell activity and the expression of basophil in children with asthma%特异性免疫治疗哮喘患儿NK细胞活性和嗜碱性细胞表达变化的影响

    Institute of Scientific and Technical Information of China (English)

    张慧娟; 董一慧; 袁显文

    2016-01-01

    目的 探讨特异性免疫治疗哮喘患儿自然杀伤细胞(natural killer cell,NK)活性和嗜碱性细胞表达变化的影响.方法 回顾性分析本院2012年12月至2014年12月收治的97例哮喘息儿的临床资料,按照治疗方法分为对照组(47例)和观察组(50例),分别给予常规治疗和特异性免疫治疗.观察两组的NK细胞活性和嗜碱性细胞表达变化,并进行比较.结果 治疗前,两组患者的NK细胞活性与嗜碱性细胞表达水平经比较差异无统计学意义(P<0.05);经过不同的治疗之后,观察组和对照组的NK细胞活性分别为(12.12±1.51)%、(10.10±2.10)%,较之本组治疗前均得到显著的提高(P<0.05);治疗后两组比较差异有统计学意义(P<0.05).观察组和对照组的嗜碱性细胞表达水平分别为(0.41±0.11)%、(0.51±0.15)%,较之本组治疗前均得到显著的下降(P<0.05);治疗后两组比较差异有统计学意义(P<0.05).结论 特异性免疫治疗哮喘患儿可以显著增强其NK细胞活性,并降低嗜碱性细胞的表达水平.%Objective To investigate the effect of specific immunotherapy on natural killer cell (NK cell) activity and the expression of basophil in asthmatic children.Methods A retrospective analysis was performed on clinical data of 97 children with asthma in our hospital from December 2013 to December 2014,and they were divided into control group (47 cases) and observation group (50 cases) according to treatment methods,and were given conventional therapy and specific immunotherapy respectively.The changes of NK cell activity and the expression of basophil in two groups were observed and compared.Results Before treatment,there were no statistically significant differences in NK cell activity and the expression of basophil between two groups (P>0.05);after treatment,NK cell activity in observation group and control group were (12.12±1.51)% and (10.10±2.10)% respectively,improved significantly compared

  3. Analysis of atypical lymphocyte and basophilic granulocyte warning information indicated by Sysmex XE-5000 blood cell analyzer%Sysmex XE-5000血细胞分析仪对异型淋巴细胞和嗜碱性粒细胞的报警提示分析

    Institute of Scientific and Technical Information of China (English)

    王凤; 刘爽; 万丽平; 韩晓丹; 王贞

    2015-01-01

    目的:评价Sysmex XE-5000血细胞分析仪对异型淋巴细胞和嗜碱性粒细胞的报警提示功能。方法收集Sysmex XE-5000血细胞分析仪同时提示异型淋巴细胞和嗜碱性粒细胞的外周血标本197份,以及单独提示异型淋巴细胞的外周血标本914份,进行显微镜复检,以核实报警提示信息是否正确。结果以显微镜复检结果为金标准,同时提示异型淋巴细胞和嗜碱性粒细胞增多的标本经复检后嗜碱性粒细胞在正常范围内,异型淋巴细胞检出的符合率为64.9%;单独提示异型淋巴增多的标本经复检后符合率为72.5%,前者明显低于后者,差异有统计学意义(P<0.05)。结论 Sysmex XE-5000血细胞分析仪同时报警提示异型淋巴细胞和嗜碱性粒细胞增多时,两者计数有一定相互干扰,需结合显微镜复检,以降低漏诊和误诊概率。%Objective To evaluate the performance of the Sysmex XE-5000 analyzer for analyzing atypical lymphocytes ,baso-philic granulocytes and their abnormalities warnings .Methods A total of 197 specimens with both atypical lymphocytes and baso-philic granulocytes warnings and 914 specimens with single warning of atypical lymphocytes indicated by Sysmex-5000 blood cell analyzer were collected and inspected by microscope simultaneously .Results Using microscopic examination as a standard ,baso-philic granulocytes within the normal range ,the coincidence rate of samples with both atypical lymphocytes and basophilic granulo-cytes warnings was 64 .9% ,while the coincidence rate of samples with single warning of atypical lymphocytes was 72 .5% .The for-mer was significantly lower than the latter(P<0 .05) .Conclusion When Sysmex XE-5000 indicates atypical lymphocytes and baso-philic granulocytes simultaneously ,there is interference between each other .It should be combined with microscopic examination in order to reduce the probability of missed diagnosis and

  4. Histamine release from cord blood basophils

    DEFF Research Database (Denmark)

    Nielsen, Bent Windelborg; Damsgaard, Tine Engberg; Herlin, Troels

    1990-01-01

    The histamine release (HR) after challenge with anti-IgE, concanavalin A, N-formyl-met-leu-phe and the calcium ionophore A23187 from 97 cord blood samples was determined by a microfiber-based assay. Maximum HR with anti-IgE showed great inter-individual variation (median: 20.5; range: 1-104 ng...

  5. Test de activación de basófilos en el diagnóstico de alergia a medicamentos Basophil activation test in the diagnosis of allergy to medicines

    Directory of Open Access Journals (Sweden)

    M.L. Sanz

    2003-01-01

    Full Text Available En el presente trabajo se estudia la fiabilidad del test de activación de basófilos (TAB en el diagnóstico "in vitro" de alergia a betalactámicos y a metamizol y se analiza la sensibilidad y especificidad de la técnica. Para ello se estudian 58 pacientes alérgicos a antibióticos betalactámicos con prueba cutánea positiva frente a algún derivado de la penicilina y 30 controles sanos que toleraban betalactámicos, y 26 pacientes alérgicos a metamizol con una reacción inmediata y 30 controles sanos que toleraban el medicamento. La sensibilidad del TAB en alergia a betalactámicos fue del 52,8% y la especificidad del 92,6%. Para metamizol la sensibilidad fue del 42,3% y la especificidad del 100%. El valor predictivo positivo del TAB en alergia a betalactámicos fue del 18,9% y el valor predictivo negativo del 98,4%. Para metamizol el valor predictivo positivo de la técnica fue del 100% y el valor predictivo negativo del 99,4%. La utilización conjunta de TAB y CAP (IgE específica permite diagnosticar un 65% de los pacientes alérgicos a betalactámicos. El uso combinado de pruebas cutáneas y TAB en alergia a metamizol detecta el 70% de los casos. El TAB es una técnica útil, no invasiva en el diagnóstico "in vitro" de la alergia a betalactámicos y metamizol.In this paper we study the reliability of the basophil activation test (BAT in the "in-vitro" diagnosis of allergy to betalactams and to metamizol, and the sensitivity and specificity of the technique are analyzed. To this end, we studied 58 patients allergic to betalactam antibiotics with a positive cutaneous test facing any derivative of penicillin and 30 healthy controls who tolerated betalactams, and 26 patients allergic to metamizol with an immediate reaction and 30 healthy controls who tolerated the medicine. Sensitivity to BAT in allergy to betalactams was 52.8%, and specificity was 92.6%. For metamizol, sensitivity was 42.3% and specificity was 100%. The positive

  6. The number of FceRI receptors on basophils decreases during subcutaneous immunotherapy

    DEFF Research Database (Denmark)

    Schmid, J. M.; Dahl, R.; Hoffmann, H. J.

    2015-01-01

    Background: Allergen specific immunotherapy is the only disease modifying treatment of allergic diseases. It induces complex cellular and humoral changes leading to an inhibition of type-1 allergic reactions. Method: Twenty four young grass pollen allergic adults suffering from seasonal rhino-con...

  7. Rat basophilic leukemia cells express syntaxin-3 and VAMP-7 in granule membranes.

    Science.gov (United States)

    Hibi, T; Hirashima, N; Nakanishi, M

    2000-04-29

    In neuronal cells, it is generally agreed that SNARE proteins underlie the release of neurotransmitter. It is controversial, however, whether they also work functionally in the degranulation of RBL-2H3 cells because the expression of SNARE proteins has not been confirmed and the degranulation is not inhibited by tetanus toxin which cleaves one of SNARE proteins, VAMP-2. We investigated the expression and the localization of SNARE proteins including VAMP-7 which is insensitive to tetanus toxin. RT-PCR analysis showed the existence of SNARE proteins, including syntaxin-2, -3, -4, SNAP-23, VAMP-2, and VAMP-7. Experiments using GFP-conjugated proteins revealed that VAMP-7 was localized only in granule membranes, whereas syntaxin-3 was in both the plasma and granule membranes. Upon antigen stimulation, these proteins in granule membranes moved to the cell surface due to the fusion of granules with the plasma membrane. The results suggest the involvement of SNARE proteins in the degranulation of RBL-2H3 cells.

  8. Effect of immunotherapy on basophil activation induced by allergens in patients with atopic dermatitis.

    Science.gov (United States)

    Sánchez, Jorge; Cardona, Ricardo

    2014-01-01

    Antecedentes: la inmunoterapia subcutánea con alergenos ha demostrado ser sumamente efectiva para el tratamiento de las enfermedades respiratorias mediadas por IgE. Sin embargo, pocos estudios exploran los mecanismos inmunológicos de la inmunoterapia en pacientes con dermatitis atópica. Objetivo: explorar la respuesta inmunológica en pacientes con dermatitis atópica que reciben inmunoterapia con ácaros de acuerdo con la inmunidad humoral y la activación de basófilos. Material y método: estudio abierto en el que se evaluó la severidad de la dermatitis con el índice SCORAD en 20 pacientes (10 con inmunoterapia y 10 sin inmunoterapia) cada tres meses durante dos años. Las muestras de suero se tomaron previo al inicio del estudio y al primer y segundo año de seguimiento para evaluar la expresión de CD63 en basófilos, concentraciones de IgE total, IgE e IgG4 específica para Der p y Der f. Diez pacientes con rinitis alérgica y cinco controles no alérgicos se incluyeron en el estudio como controles. Resultados: la expresión de CD63 en los basófilos después de la estimulación con Der p fue más alta en los pacientes con dermatitis que en los pacientes con rinitis y en los sujetos no alérgicos. Luego del primer y segundo año de tratamiento, la expresión de CD63 fue menor en el grupo de pacientes con dermatitis que recibieron inmunoterapia en comparación con los tres grupos control. Observamos una correlación entre el SCORAD, IgG4 y la expresión de CD63. Conclusiones: en pacientes con dermatitis, la prueba de activación de basófilos podría usarse como biomarcador de respuesta clínica; asimismo, la modulación de esta célula puede llevar a un mejor control clínico.

  9. [Modified tests basophil degranulation and leukocyte migration inhibition factor in drug allergy. Study 2009-2014].

    Science.gov (United States)

    Irigoyen-Coria, María Lourdes; Rojo-Gutiérrez, María Isabel; Meyer-Gómez, Rubén Humberto; Leyva-Carmona, Isabel; Zendejas-Buitrón, Víctor Manuel; García-Ruiz, Angélica Dinorah; Mellado-Ábrego, Jaime; García-Ortega, Ilayalid; Castillo-Narváez, Gloria

    2016-01-01

    Antecedentes: Existe incremento de reacciones adversas a medicamentos y pocos estudios para el diagnóstico. Objetivo: Determinar la utilidad de pruebas modificadas de degranulación de basófilos (DB) y del factor inhibidor de la migración de leucocitos (LIF, leukocyte migration inhibition factor) para comprobar la hipersensibilidad a medicamentos. Métodos: Se estudiaron 177 pacientes, de uno y otro sexo, con diagnóstico de hipersensibilidad a medicamentos, en quienes se determinó pruebas modificadas de DB, LIF, o ambas entre 2009 y 2014. Se parearon con controles positivos, negativos y población no alérgica. Las solicitudes se emitieron de acuerdo con el tipo de hipersensibilidad, considerando tipo I a DB y tipo IV a LIF Resultados: 170 pacientes (96.04%) fueron positivos al menos a un medicamento (RR, 4.71). Se realizaron 561 pruebas modificadas de DB (73.62%) y 201 de LIF (26.37%). El sexo femenino fue más frecuente (64.41%); la edad promedio fue de 38.5 años. La prueba modificada de DB resultó positiva en 70.23% y la de LIF en 67.16%. La sensibilidad de las pruebas se incrementó en forma complementaria y a dos diluciones. La correlación de las pruebas fue altamente significativa. Conclusiones: Las mujeres presentan más reacciones a fármacos. La prueba modificada de DB es útil en cualquier edad. Como los medicamentos pueden activar uno u otro mecanismo de hipersensibilidad es importante solicitar las pruebas simultáneamente.

  10. Allergic rhinitis is a local disease: the role of local IgE production, basophils and mast cells

    NARCIS (Netherlands)

    A. Kleinjan (Alex)

    2002-01-01

    textabstractThe introduction to this thesis summarizes the literature which indicates that there is a discrepancy between sensitisation and allergic disease. Two aspects which might play a role in this discrepancy are the differences between production and funtion of local versus systemic lgE and th

  11. Suppression of Basophil Histamine Release and Other IgE-dependent Responses in Childhood Schistosoma mansoni/hookworm Coinfection

    OpenAIRE

    Pinot de Moira, Angela; Fitzsimmons, Colin M.; Frances M Jones; Wilson, Shona; Cahen, Pierre; Tukahebwa, Edridah; Mpairwe, Harriet; Mwatha, Joseph K; Bethony, Jeffrey M.; Skov, Per S.; Kabatereine, Narcis B.; Dunne, David W.

    2014-01-01

    Background.  The poor correlation between allergen-specific immunoglobulin E (asIgE) and clinical signs of allergy in helminth infected populations suggests that helminth infections could protect against allergy by uncoupling asIgE from its effector mechanisms. We investigated this hypothesis in Ugandan schoolchildren coinfected with Schistosoma mansoni and hookworm. Methods.  Skin prick test (SPT) sensitivity to house dust mite allergen (HDM) and current wheeze were assessed pre-anthelmintic...

  12. Anti-degranulating activity in rat basophil leukemia RBL-2H3 cells of flavanone glycosides and their aglycones in citrus fruits.

    Science.gov (United States)

    Murata, Kazuya; Takano, Seiya; Masuda, Megumi; Iinuma, Munekazu; Matsuda, Hideaki

    2013-07-01

    The anti-degranulating activity of flavonoids present in Citrus fruits was comprehensively evaluated. Among these, hesperetin and naringenin, respectively aglycones of hesperidin and narirutin, showed significant activity. The targets of hesperetin and naringenin were found: hesperetin inhibited phosphorylation of Syk and Akt, while naringenin suppressed the expression of Lyn and inhibited the phosphorylation of Akt. These results suggest that hesperetin and naringenin inhibit degranulation by suppression of pathway signals and reduce the symptoms of allergy by inhibiting phosphorylation of Akt, which leads to the suppression of cytokines. In addition, hesperetin showed inhibitory activity against the degranulation induced by calcium ionophores, indicating that hesperetin exerts its inhibitory activity by stabilizing the membrane structure.

  13. Structural analysis of ARC-type inhibitor (ARC-1034) binding to protein kinase A catalytic subunit and rational design of bisubstrate analogue inhibitors of basophilic protein kinases.

    Science.gov (United States)

    Lavogina, Darja; Lust, Marje; Viil, Indrek; König, Norbert; Raidaru, Gerda; Rogozina, Jevgenia; Enkvist, Erki; Uri, Asko; Bossemeyer, Dirk

    2009-01-22

    The crystal structure of a complex of the catalytic subunit (type alpha) of cAMP-dependent protein kinase (PKA C alpha) with ARC-type inhibitor (ARC-1034), the presumed lead scaffold of previously reported adenosine-oligo-arginine conjugate-based (ARC-type) inhibitors, was solved. Structural elements important for interaction with the kinase were established with specifically modified derivatives of the lead compound. On the basis of this knowledge, a new generation of inhibitors, conjugates of adenosine-4'-dehydroxymethyl-4'-carboxylic acid moiety and oligo(D-arginine), was developed with inhibitory constants well into the subnanomolar range. The structural determinants of selectivity of the new compounds were established in assays with ROCK-II and PKBgamma.

  14. Structural analysis of the endogenous glycoallergen Hev b 2 (endo-β-1,3-glucanase) from Hevea brasiliensis and its recognition by human basophils

    OpenAIRE

    Rodríguez-Romero, Adela; Hernández-Santoyo, Alejandra; Fuentes-Silva, Deyanira; Palomares, Laura A.; Muñoz-Cruz, Samira; Yépez-Mulia, Lilian; Orozco-Martínez, Socorro

    2014-01-01

    This study describes the three-dimensional structure of the endogenous glycosylated allergen Hev b 2 (endo-β-1,3-glucanase), which exhibits three post-translational modifications that form a patch on the surface of the molecule that is proposed to be an allergenic IgE epitope.

  15. Eotaxin-2, a Novel CC Chemokine that Is Selective for the Chemokine Receptor CCR3, and Acts Like Eotaxin on Human Eosinophil and Basophil Leukocytes

    OpenAIRE

    Forssmann, Ulf; Uguccioni, Mariagrazia; Loetscher, Pius; Dahinden, Clemens A; Langen, Hanno; Thelen, Marcus; Baggiolini, Marco

    1997-01-01

    A novel human CC chemokine consisting of 78 amino acids and having a molecular mass of 8,778.3 daltons (VVIPSPCCMF FVSKRIPENR VVSYQLSSRS TCLKAGVIFT TKKGQQ SCGD PKQEWVQRYM KNLDAKQKKA SPRARAVA) was isolated together with three minor COOH-terminally truncated variants with 73, 75, and 76 residues. The new chemokine was termed eotaxin-2 because it is functionally very similar to eotaxin. In terms of structure, however, eotaxin and eotaxin-2 are rather distant, they share only 39% identical amino ...

  16. Inhibitory effects of whisky congeners on IgE-mediated degranulation in rat basophilic leukemia RBL-2H3 cells and passive cutaneous anaphylaxis reaction in mice.

    Science.gov (United States)

    Itoh, Tomohiro; Tsukane, Mariko; Koike, Minako; Nakamura, Chizu; Ohguchi, Kenji; Ito, Masafumi; Akao, Yukihiro; Koshimizu, Seiichi; Nozawa, Yoshinori; Wakimoto, Toshiyuki; Nukaya, Haruo; Suwa, Yoshihide

    2010-06-23

    Whisky is matured in oak casks. Many nonvolatile substances (whisky congeners, WC) seep from the oak cask during the maturing process. In this study, three antiallergic agents (syringaldehyde, SA; lyoniresinol, Lyo; and ellagic acid, EA) were isolated from WC. Treatment with SA, Lyo, and EA reduced the elevation of intracellular free Ca(2+) concentration ([Ca(2+)]i) and intracellular ROS production caused by FcepsilonRI activation. The inhibitions of the elevation of [Ca(2+)]i and intracellular ROS production by SA and Lyo were mainly due to the suppression of the NADPH oxidase activity and scavenging of the produced radical, respectively. On the other hand, EA inactivated spleen tyrosine kinase and led to the inhibition of the elevation of [Ca(2+)]i and intracellular ROS production. Furthermore, it was found that WC strongly inhibited IgE binding to the FcepsilonRIalpha chain, whereas SA, Lyo, and EA did not indicate this inhibitory effect. These results suggest that WC inhibits allergic reactions through multiple mechanisms. To disclose the in vivo effects of WC, SA, Lyo, and EA, these compounds were administered to type I allergic model mice, and the passive cutaneous anaphylaxis (PCA) reaction was measured. These compounds remarkably suppressed the PCA reaction. Taken together, these findings suggest that WC seemed to be beneficial to ameliorate allergic reactions.

  17. Microarray analysis of altered gene expression of basophils in chronic idiopathic urticaria%慢性特发性荨麻疹患者嗜碱粒细胞差异表达基因的研究

    Institute of Scientific and Technical Information of China (English)

    马一平; 姚煦; 林麟; 崔盘根

    2010-01-01

    目的:检测慢性特发性荨麻疹(CIU)患者外周血嗜碱粒细胞基因表达谱.方法:分离提取2例CIU患者和2名健康人外周血嗜碱粒细胞,提取总RNA;利用cRNA标记方法对样品进行荧光标记,然后用于芯片杂交;用LuxScan 10K/A双通道激光扫描仪进行芯片扫描;采用LuxScan 3.0图像分析软件对芯片图像进行分析;然后对芯片上的数据用Lowess方法进行归一化;最后以差异为2倍的标准来确定差异表达基因.结果:CIU患者嗜碱粒细胞与健康对照组相比存在差异表达基因共计30个.其中表达上调基因12个,表达下调基因8个,功能不明基因10个.结论:CIU患者和正常人嗜碱粒细胞的一些基因表达差异可能与CIU发病有关.

  18. Relevance of Ara h1, Ara h2 and Ara h3 in peanut-allergic patients, as determined by immunoglobulin E Western blotting, basophil-histamine release and intracutaneous testing: Ara h2 is the most important peanut allergen

    NARCIS (Netherlands)

    Koppelman, S.J.; Wensing, M.; Ertmann, M.; Knulst, A.C.; Knol, E.F.

    2004-01-01

    Background: A number of allergenic proteins in peanut has been described and the relative importance of these allergens is yet to be determined. Objectives: We have investigated the relevance of previously identified peanut allergens in well-characterized peanut-allergic patients by in vitro, ex viv

  19. 気管支喘息における好塩基球の動態に関する研究 第1編フローサイトメトリーを用いたヒト末梢血好塩基球の高純度分離法について) 第2編各種サイトカインによるヒト好塩基球の遊走活性に関する検討)

    OpenAIRE

    谷本, 安

    1993-01-01

    1.Basophils were purified from peripheral blood of normal donors using Percoll discontinuous gradients and negative selection by flow cytometry. The mean purity of basophils obtained was 84.7 +/- 4.1 (s.d.)% (range 77.3-90.0%, n = 13). The overall yield of these procedures was 16.0 +/- 2.6% (range 11.0-19.9%, n = 13), and cell viability of purified basophils exceeded 90%. Properties of highly purified basophils obtained by flow cytometry did not differ from those of partially enriched basophi...

  20. Topical ophthalmic beta blockers may cause release of histamine through cytotoxic effects of inflammatory cells

    NARCIS (Netherlands)

    Beek, van L.M.; Mulder, M.; Haeringen, van N.J.; Kijlstra, A.

    2000-01-01

    Aim - To evaluate the effects of β blockers used in ophthalmology on the release of histamine from mixed cell preparations containing human leucocytes and basophils. Methods - A mixed leucocyte and basophil preparation was obtained from venous blood of healthy non-atopic volunteers. Cell preparation

  1. Flow cytometry analysis of basophil degranulation in allergy to dust mite%流式细胞术分析户尘螨致敏的嗜碱性粒细胞活化试验及其临床意义

    Institute of Scientific and Technical Information of China (English)

    徐翀; 王小蕊; 蒋黎敏; 王连升; 袁向亮; 潘秀军; 沈立松

    2007-01-01

    目的 建立流式细胞术(FCM)分析嗜碱性粒细胞活化试验(BAT)的方法并评估其在过敏性疾病诊断中的意义.方法 以CD63/CD203c/CD45抗体组合,建立用FCM分析户尘螨致敏的嗜碱性粒细胞脱颗粒的方法.以皮肤点刺试验(SPT)作为金标准方法,将FCM检测活化嗜碱性粒细胞与荧光酶联免疫吸附试验(FELISA)测定特异性IgE(sIgE)结果作比较,分析其临床应用价值.结果 以CD45和CD203c联合设门,可以得到纯的嗜碱性粒细胞;CD63是活化嗜碱性粒细胞的最佳标志;Spearman相关系数显示,sIgE的Unicap分级与活化嗜碱性粒细胞呈中度相关;FCM检测活化嗜碱性粒细胞与FELISA测定sIgE结果在户尘螨过敏性疾病诊断中的差异无统计学意义(P>0.05),但前者的敏感度、符合率、阳性预测值以及阳性似然比各项指标都优于后者.结论 通过FCM分析CD63的表达来确定活化嗜碱性粒细胞是诊断速发型超敏反应的一种有效、安全的体外检测方法.

  2. 荨麻疹患者外周血嗜碱性粒细胞表面CD63表达的研究%Analysis of CD63 expression on basophils from human peripheral blood of the patients with urticaria

    Institute of Scientific and Technical Information of China (English)

    李振; 肇静娴; 蔡小嫦; 邓列华

    2008-01-01

    目的 观察荨麻疹患者外周血中活化的嗜碱性粒细胞表面表达CD63的数量,探讨嗜碱性粒细胞的活化程度,为临床检测提供新方法.方法 用屋尘螨浸液作划破试验,筛选敏感的荨麻疹患者.用CD63-FITC等抗体标记嗜碱性粒细胞,流式细胞技术检测CD63的表达情况.结果 急、慢性患者活化嗜碱性粒细胞百分率分别为33.77~15.75%、11.66~5.25%,与正常人比较均有显著差异(P<0.05);经变应原刺激后,急、慢性患者活化嗜碱性粒细胞百分率分别为60.25~18.88%、22.22~7.89%,急、慢性患者活化前、活化后也均存在明显差异(P<0.05),而且急性患者活化程度更高(P<0.05).结论 通过测定嗜碱性粒细胞表达CD63可能有助于过敏的诊断.

  3. HIV-1 envelope gp41 peptides promote migration of human Fc epsilon RI+ cells and inhibit IL-13 synthesis through interaction with formyl peptide receptors.

    Science.gov (United States)

    de Paulis, Amato; Florio, Giovanni; Prevete, Nella; Triggiani, Massimo; Fiorentino, Isabella; Genovese, Arturo; Marone, Gianni

    2002-10-15

    We evaluated the effects of synthetic peptides (2017, 2019, 2020, 2021, 2023, 2027, 2029, 2030, 2031, and 2035) encompassing the structure of HIV-1(MN) envelope gp41 on both chemotaxis of human basophils and the release of preformed mediators (histamine) and of cytokines (IL-13). Peptides 2019 and 2021 were potent basophil chemoattractants, whereas the other peptides examined were ineffective. Preincubation of basophils with FMLP or gp41 2019 resulted in complete desensitization to a subsequent challenge with homologous stimulus. Incubation of basophils with low concentration (5 x 10(-7) M) of FMLP, which binds with high affinity to N-formyl peptide receptor (FPR), but not to FPR-like 1, did not affect the chemotactic response to a heterologous stimulus (gp41 2019). In contrast, a high concentration (10(-4) M) of FMLP, which binds also to FPR-like 1, significantly reduced the chemotactic response to gp41 2019. The FPR antagonist cyclosporin H inhibited chemotaxis induced by FMLP, but not by gp41 2019. None of these peptides singly induced the release of histamine or cytokines (IL-4 and IL-13) from basophils. However, low concentrations of peptides 2019 and 2021 (10(-8)-10(-6) M) inhibited histamine release from basophils challenged with FMLP but not the secretion caused by anti-IgE and gp120. Preincubation of basophils with peptides 2019 and 2021 inhibited the expression of both IL-13 mRNA, and the FMLP-induced release of IL-13 from basophils. These data highlight the complexity of the interactions between viral and bacterial peptides with FPR subtypes on human basophils.

  4. A novel human immunoglobulin Fcγ–Fcε bifunctional fusion protein inhibits FcεRI-mediated degranulation

    OpenAIRE

    Zhu, Daocheng; Kepley, Christopher L.; Zhang, Min; Zhang, Ke; Saxon, Andrew

    2002-01-01

    Human mast cells and basophils that express the high-affinity immunoglobulin E (IgE) receptor, Fcε receptor 1 (FcεRI), have key roles in allergic diseases. FcεRI cross-linking stimulates the release of allergic mediators1. Mast cells and basophils co-express FcγRIIb, a low affinity receptor containing an immunoreceptor tyrosine-based inhibitory motif and whose co-aggregation with FcεRI can block FcεRI-mediated reactivity2–4. Here we designed, expressed and tested the human basophil and mast-c...

  5. Retinoblastoma: genetic considerations and report of a new animal model

    NARCIS (Netherlands)

    Albert, D.M.; O'Brien, J.M.; Marcus, D.M.; Bernards, R.A.

    1990-01-01

    Retinoblastoma is the most common primary, intraocular neoplasm of childhood. Histologically, retinoblastoma resembles, in many respects, other pediatric malignancies such as medulloblastoma and neuroblastoma. These tumors are composed of small, basophilic cells with scanty cytoplasm and often form

  6. Development of Chemiluminscence Immunoaasy Kit for Follicle-Stimulating Hormone

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>Follicle-stimulating hormone (FSH) is a gonadotropic hormone, and it is synthesized and secreted by basophilic cell of anterior lobe of hypophysis. Detection of FSH levels in human serum is useful in

  7. Expression profiling of constitutive mast cells reveals a unique identity within the immune system

    Science.gov (United States)

    Dwyer, Daniel F.; Barrett, Nora A.; Austen, K. Frank

    2016-01-01

    Mast cells are evolutionarily ancient sentinel cells. Like basophils, mast cells express the high-affinity IgE receptor and are implicated in host defense and diverse immune-mediated diseases. To better characterize the function of these cells, we assessed the transcriptional profiles of mast cells isolated from peripheral connective tissues and basophils isolated from spleen and blood. We found that mast cells were transcriptionally distinct, clustering independently from all other profiled cells, and that mast cells demonstrated considerably greater heterogeneity across tissues than previously appreciated. We observed minimal homology between mast cells and basophils, which share more overlap with other circulating granulocytes than with mast cells. Derivation of mast cell and basophil transcriptional signatures underscores their differential capacity to detect environmental signals and influence the inflammatory milieu. PMID:27135604

  8. Structural, cytochemical, immunocytochemical and ultrastructural characterization of blood granulocytes of the roadside hawk Buteo magnirostris (Gmelin, 1788) (Avian, Falconiform).

    Science.gov (United States)

    Santos, A A; Da Silva, A Marques Joppert; Silva, M R Regis; Segreto, H R Cômodo; Egami, M Imoto

    2003-10-01

    In the peripheral blood of the roadside hawk, Buteo magnirostris, the following types of granulocytic leucocytes were identified: heterophil, eosinophil and basophil. The heterophils presented acidophilic and spindle shaped granules, the eosinophils possess spherical eosinophilic granules and the basophils showed spherical and basophilic granules. The heterophils and eosinophils presented positive cytochemical reaction to glycogen and basic polyaminoacid, while the eosinophils presented sudanophilic granules, which were also positive for myeloperoxidase. The heterophils, alone, presented positivity for acid phosphatase in some granules and immunoreactivity to TGF-beta1 was observed only in the cytoplasm of the eosinophils. Electron microscopy demonstrated the heterophil granules as predominantly spindle shaped, being strongly electron-dense, while the eosinophils had numerous uniformly electron-dense spherical granules and the basophils presented three different types of granules identified according to their electron-density and the aspect of their matrix.

  9. AN OVACYSTIS-LIKE CONDITION IN THE AMERICAN OYSTER CRASSOSTREA VIRGINICA GMELIN FROM THE NORTHEASTERN GULF OF MEXICO

    Science.gov (United States)

    Histological examination of the eastern oyster, Crassostrea virginica, from a study in Pensacola Bay, Florida, revealed two cases of abnormally large, basophilic ova that resemble ovacystis disease previously reported in oysters from Maine and Long Island. The hypertrophied gamet...

  10. Use of a new glass microfibre histamine release method to study the modulation of the host response in human schistosomiasis mansoni. Individuals with different degrees of exposure to the disease show differing antibody biological function

    DEFF Research Database (Denmark)

    Satti, M.Z.; Ebbesen, F.; Vennervald, B.;

    1996-01-01

    A new glass microfibre histamine release method was used to study the modulation of the host response in human schistosomiasis mansoni to improve our understanding of the role of basophils in the development of immunity in schistosomiasis mansoni. The histamine release from umbilical cord blood b...... basophils sensitized with sera from Sudanese individuals infected with Schistosoma mansoni was measured. Schistosomiasis sera (n=113) were able to passively sensitize basophils and induce a positive histamine release in response to whole worm homogenate (WWH)(chi(2)=40.5, P......A new glass microfibre histamine release method was used to study the modulation of the host response in human schistosomiasis mansoni to improve our understanding of the role of basophils in the development of immunity in schistosomiasis mansoni. The histamine release from umbilical cord blood...

  11. Loxoprofen sodium induces the production of complement C5a in human serum.

    Science.gov (United States)

    Kumagai, Tomoaki; Yamaguchi, Nozomi; Hirai, Hiroyuki; Kojima, Shigeyuki; Kodani, Yoshiko; Hashiguchi, Akihiko; Haida, Michiko; Nakamura, Masataka

    2016-04-01

    Basophil activation test (BAT) is an in vitro allergy test that is useful to identify allergens that cause IgE-dependent allergies. The test has been used to detect not only food allergies and allergies caused by environmental factors but also to detect drug hypersensitivity, which has been known to include IgE-independent reactions. In our preliminary studies in which BAT was applied to detect hypersensitivity of loxoprofen, a non-steroidal anti-inflammatory drug (NSAID), conventional BAT with incubation for 30min did not show basophil activation by means of increased CD203c expression. In this study, we extended the incubation time to 24h on the basis of the hypothesis that loxoprofen indirectly activates basophils. Basophils from healthy control donors as well as allergic patients showed up-regulation of CD203c after incubation with loxoprofen for 24h. Activation was induced using loxoprofen-treated serum. Proteomic and pharmacologic analyses revealed that serum incubation with loxoprofen generated an active complement component C5a, which induced CD203c expression via binding to the C5a receptor on basophils. Because C3a production was also detected after incubation for 24h, loxoprofen is likely to stimulate the complement classical pathway. Our findings suggest that the complement activation is involved in drug hypersensitivity and the suppression of this activation may contribute to the elimination of false positive of BAT for drug allergies.

  12. Assessment of chronic spontaneous urticaria by serum-induced tumor necrosis factor alpha and matrix metalloproteinase-9 release

    DEFF Research Database (Denmark)

    Falkencrone, Sidsel; Bindslev-Jensen, Carsten; Skov, Per Stahl;

    isolated with MACS Basophil Isolation Kit to 97-99% purity. Cells were pulsed 1 hour with/without anti-IgE or with sera from CSU-patients/healthy controls and incubated for a total of 21 h before protein analysis of supernatants. MMP-9 and TNFα in supernatants were measured with commercial ELISAs (R......BACKGROUND Previous studies from our group have demonstrated that IgE-mediated basophil activation leads to release of TNFα that in turn can induce matrix metallo-proteinase-9 (MMP-9) release from monocytes. We wished to investigate if serum from chronic spontaneous urticaria-patients with auto......-antibodies against IgE/IgE-receptor could induce TNFα and MMP-9 release from donor PBMCs, and if release levels could be used to assess severity and activity of chronic spontaneous urticaria (CSU). METHODS Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood from healthy donors and basophils...

  13. Experimental Researches on Inhibitory Effect of Huoxiang Zhengqi Liquid(藿香正气水) on Histamine Release

    Institute of Scientific and Technical Information of China (English)

    余传星; 朱玲

    2003-01-01

    Objective: To observe the blocking effect of Huoxiang Zhengqi liquid (藿香正气水,HXZQL) on histamine release of basophils. Methods: The study was conducted using HXZQL to neutralize and block IgE and antagonize the effect of IL-3 induced enhancing histamine release of bosaphils. Concentration and percentage of histamine release of bosaphils were tested by the basophil histamine release test. Results: The HXZQL drug-serum had obvious inhibitive effect on degranulation of bosaphils induced by IgE antigen-antibody complex (P<0.05) and can successfully weaken the histamine release of basophils stimulated by IL-3. When compared with those in the untreated group, it showed P<0.01. Conclusion: HXZQL drug-serum may neutralize and block IgE and has inhibitive effects on degranulation of cells when resensitized by antigens. It can also block allergic response type Ⅰ by antagonizing the effect of IL-3 induced enhancing histamine release.

  14. Histamine immunocytochemistry

    DEFF Research Database (Denmark)

    Jacobi, H H; Johansson, O; Liang, Y

    2000-01-01

    We report that basophils in peripheral blood can be stained using histamine immunocytochemistry. The staining is based on the fixation of leucocytes with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (CDI) and the subsequent incubation of these cells with antisera raised against histamine...... conjugated to different carrier proteins using CDI. The staining appears to be specific for basophils and stained cells can be examined using both fluorescence microscopy and flow cytometry. In addition, histamine immunocytochemistry can be combined with conventional immunocytochemistry by incubating...... leucocytes with antibodies to cell surface antigens prior to or following fixation of the cells with CDI. Thus, histamine immunocytochemistry may be a valuable tool in future studies of human basophils....

  15. [Histamine releasing activity of dental materials as the indicator of their biocompatibility].

    Science.gov (United States)

    Babakhin, A A; Volozhin, A I; Dubova, L V; Lebedenko, I Iu; Babakhina, Iu A; Zhuravleva, A A; Diubuske, L M

    2008-01-01

    Different types of dental materials (DM) were studied for their capacity to release histamine in vitro from basophils of whole blood of allergic patients and healthy donors using automated and computerized glass fiber-based leukocyte histamine release test (LHRT). It was shown that some types of DM possessed ability to release histamine from basophils and some didn't. There were no differences in histamine releaseability from basophils obtained from allergic patients and healthy donors. LHRT gives opportunity to recognize of DM possessing high or low histamine releaseability as well as to detect individual sensitivity to different DM. Thus, LHRT can be used for preliminary assessment of DM for their biocompatibility and also for individual selection of suitable DM for particular patient to avoid unwanted side effects.

  16. Reversible alterations in epithelial cell turnover in digestive gland of winkles (Littorina littorea) exposed to cadmium and their implications for biomarker measurements.

    Science.gov (United States)

    Zaldibar, B; Cancio, I; Marigómez, I

    2007-02-28

    In marine molluscs, the epithelium of the digestive gland is composed of two cell types, namely, digestive and basophilic cells. Under normal physiological conditions digestive cells outnumber basophilic cells, but under different stress situations the composition of the epithelium changes, basophilic cells apparently replace digestive cell. Winkles, Littorina littorea, were exposed to 1.25mg/l Cd for 20 days to provoke cell type replacement. Then, animals were depurated in clean seawater for 10 days to determine whether cell type replacement was reversible. Digestive glands were fixed in Carnoy and paraffin embedded for histological analysis. The volume densities of basophilic cells (Vv(BAS)) and digestive cells (Vv(DIG)) were calculated by stereology on hematoxylin-eosin stained sections. Vv(BAS) increased and Vv(DIG) decreased in Cd-exposed animals. After estimation of cell size and absolute cell numbers, these changes were attributed to digestive cell loss and concomitant basophilic cell hypertrophy but not to increased numbers of basophilic cells. Cell type composition and cell size almost fully returned to normal values after 10-day depuration. Accordingly, PCNA immunohistochemistry demonstrated that proliferating digestive cells were more abundant in winkles exposed to Cd and after 10-day depuration than in control specimens, suggesting that net digestive cell loss was accompanied by increased digestive cell proliferation. Thus, Cd-exposure seems to provoke an enhanced digestive cell turnover in order to cope with Cd detoxification. Intralysosomal accumulation of metals (autometallographied black silver deposits; BSD) was used as a biomarker of exposure to Cd and lysosomal structural changes as an effect biomarker to see whether cell type composition might have any effect on these endpoints. BSD formed around Cd ions, in digestive cell lysosomes of Cd-exposed winkles whereas basophilic cells appeared devoid of them. After depuration, BSD were less

  17. Acute lead poisoning in two users of illicit methamphetamine

    Energy Technology Data Exchange (ETDEWEB)

    Allcott, J.V. III; Barnhart, R.A.; Mooney, L.A.

    1987-07-31

    Acute lead poisoning can present a difficult diagnostic dilemma, with symptoms that mimic those of hepatitis, nephritis, and encephalopathy. The authors report two cases in intravenous methamphetamine users who presented with abnormal liver function values, low hematocrit values, basophilic stippling of red blood cells, and elevated blood lead levels. Both patients excreted large amounts of lead in their urine after treatment with edetic acid, followed by resolution of their symptoms. Lead contamination was proved in one drug sample. Basophilic stippling of the red blood cells was the one key laboratory result that led to the definitive diagnosis in both cases.

  18. The immunoglobulin superfamily member CD200R identifies cells involved in type 2 immune responses

    DEFF Research Database (Denmark)

    Blom, Lars H; Martel, Britta C; Larsen, Lau F

    2017-01-01

    BACKGROUND: The pathology of allergic diseases involves type 2 immune cells, such as Th2, ILC2, and basophils exerting their effect by production of IL-4, IL-5, and IL-13. However, surface receptors that are specifically expressed on type 2 immune cells are less well documented. The aim...... and ILC2 cells and basophils. In peanut-allergic subjects the peanut-specific Th2 (CD154(+) CRTh2(+) ) cells expressed more CD200R than the non-allergen specific Th2 (CD154(-) CRTh2(+) ) cells. Moreover, co-staining of CD161 and CD200R identified peanut-specific highly differentiated IL-4(+) IL-5(+) Th2...

  19. Immunochemical and biological quantification of peanut extract

    DEFF Research Database (Denmark)

    Poulsen, Lars K.; Pedersen, Mona H; Platzer, Michael;

    2003-01-01

    Biological standardization of allergen extracts is one of the steps in the characterization of an extract. The gold standard for determination of biological potency is the skin prick test, but histamine release (HR) has been used as a convenient ex vivo method for analyzing a large number...... of samples. We describe the use of rabbit basophils as a tool in biological standardization. Using peanut as a model allergen, it is described how rabbits immunized for production of antiserum may become sensitized and their basophils used for histamine release experiments. It is also possible to use rabbit...

  20. “Preleukemic or smoldering” chronic myelogenous leukemia (CML:BCR-ABL1 positive: A brief case report

    Directory of Open Access Journals (Sweden)

    John M. Bennett

    2015-01-01

    The most common feature of CML is an elevated WBC count, usually above 25×103/µL, and frequently above 100×103/µL. We report a case of confirmed Ph+CML with a normal CBC detected because of the presence of rare myelocytes and 2% basophils [Fig. 1]. Previous leukocyte counts for the preceding eight years were normal with the exception of one done four months prior to his presentation that showed an abnormal differential with 1% basophils, 2% metamyelocytes and 2% myelocytes.

  1. Mast Cell Growth Factor Enhances Multilineage Hematopoietic Recovery in Vivo Following Radiation-Induced Aplasia

    Science.gov (United States)

    1994-01-01

    lymphocyte, monocyte, tributing to morbidity and mortality associated with hemato- eosinophil , and basophil numbers, as well as an increase in...factor (ligand for c-kit) administered in murine mast cell growth factor (c-kit figand) on colony vivo to mice either alone or in combination with granu

  2. The Pathophysiology of Combined Radiation Injuries: A Review and Analysis of the Literature on Non-Human Research

    Science.gov (United States)

    1991-07-01

    21 when RIO is given first probably was the result of greatly reduced number of granu - locytes and macrophages caused by earlier radiation-induced bone...macrophage colony stimulating factor. GRANULOCYTES. White blood cells containing neutrophil, basophil or eosinophil granules in the cytoplasm. GRAY (Gy). Unit

  3. Generation and epitope analysis of human monoclonal antibody isotypes with specificity for the timothy grass major allergen Phl p 5a

    DEFF Research Database (Denmark)

    Hecker, J.; Diethers, A.; Seismann, H.;

    2011-01-01

    of the antibodies with the allergen was assessed. Applicability in allergy diagnostics was confirmed by establishment of artificial human sera. Functionality of both antibodies was further demonstrated in receptor binding studies and mediator release assays using humanised rat basophil leukaemia cells (RBL-SX38...

  4. Tricin from a malagasy connaraceous plant with potent antihistaminic activity.

    Science.gov (United States)

    Kuwabara, Hidenori; Mouri, Kyoko; Otsuka, Hideaki; Kasai, Ryoji; Yamasaki, Kazuo

    2003-09-01

    The bioassay-guided separation of a Malagasy plant, Agelaea pentagyna, led to the isolation of a flavonoid, tricin (1), with potent inhibitory activity toward exocytosis from antigen-stimulated rat leukemia basophils (RBL-2H3). The structure-activity relationships among structurally related natural and synthetic flavonoids are also discussed.

  5. Histochemical studies on striated muscle fibres of the beige mutant mouse

    DEFF Research Database (Denmark)

    Kirkeby, S

    1982-01-01

    A histological study of cylindric structures in skeletal muscle fibres from beige mice with the Chediak-Higashi syndrome was carried out. The muscle tissue was investigated morphologically with a differential interference contrast microscope and stained for glycogen, lipid, and basophile elements....... It is suggested that the cylindric structures represent accumulations of glycogen rich lipoprotein membranes....

  6. Doxycycline Inhibits Vascular Leakage and Prevents the Development of Pulmonary Edema

    Science.gov (United States)

    2008-12-01

    treating allergic conditions also in humans. Of the molecular structures comprising the endothelial cell-cell contacts, the adherens junctions...Grant, 1984: Histamine-releasing activity (HRA). III. HRA induces human basophil histamine release by provoking noncytotoxic granule exocytosis. Clin...tissue architecture and morphogenesis. Cell, 84, 345-57. Lampugnani, M. G. and E. Dejana, 1997: Interendothelial junctions: structure , signalling

  7. Development of β-lactoglobulin-specific chimeric human IgEκ monoclonal antibodies for in vitro safety assessment of whey hydrolysates

    NARCIS (Netherlands)

    Knipping, Karen; Simons, Peter; Buelens-Sleumer, Laura S; Cox, Linda; den Hartog, Marcel; de Jong, Niels; Teshima, Reiko; Garssen, Johan; Boon, Louis; Knippels, Léon M J

    2014-01-01

    BACKGROUND: Cow's milk-derived whey hydrolysates are nutritional substitutes for allergic infants. Safety or residual allergenicity assessment of these whey hydrolysates is crucial. Currently, rat basophilic leukemia RBL-2H3 cells expressing the human IgE receptor α-chain (huFcεRIα-RBL-2H3), sensiti

  8. A role for interleukin-33 in T(H)2-polarized intestinal inflammation?

    DEFF Research Database (Denmark)

    Seidelin, J B; Rogler, G; Nielsen, O H

    2011-01-01

    Interleukin 33 (IL-33) is a recently discovered cytokine member of the IL-1 superfamily that is widely expressed in fixed tissue cells, including endothelial and epithelial cells. IL-33 induces helper T cells, mast cells, eosinophils, and basophils to produce type-2 cytokines through binding to t...

  9. Immunoglobulin superantigen protein L induces IL-4 and IL-13 secretion from human Fc epsilon RI+ cells through interaction with the kappa light chains of IgE.

    Science.gov (United States)

    Genovese, Arturo; Borgia, Guglielmo; Björck, Lars; Petraroli, Angelica; de Paulis, Amato; Piazza, Marcello; Marone, Gianni

    2003-02-15

    Peptostreptococcus magnus protein L is a multidomain bacterial surface protein that correlates with virulence. It consists of up to five homologous Ig-binding domains (B1-B5) that interact with the variable domain of Ig kappa L chains. Intact protein L stimulates the synthesis and the release of IL-4 and IL-13 from human basophils in vitro. A protein L fragment covering the Ig-binding domains B1-B4 also induced IL-4 and IL-13 release from basophils. There was an excellent correlation (r(s) = 0.82; p ADZ (kappa chains) blocked both anti-IgE- and protein L-induced secretion. Cyclosporin A, but not cyclosporin H, inhibited protein L-induced release of IL-4 and IL-13 from basophils. Thus, protein L acts as a bacterial Ig superantigen to induce the synthesis and release of IL-4 and IL-13 from basophils by interacting with kappa L chains of the IgE isotype.

  10. Novel Strategy to Create Hypoallergenic Peanut Protein-Polyphenol Edible Matrices for Oral Immunotherapy

    Science.gov (United States)

    Peanut allergy is an IgE-mediated hypersensitivity. Upon peanut consumption by an allergic individual, epitopes on peanut proteins bind and cross-link peanut-specific IgE on mast cell and basophil surfaces triggering the cells to release inflammatory mediators responsible for allergic reactions. P...

  11. Calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus)

    Institute of Scientific and Technical Information of China (English)

    Chisoni Mumba; David Squarre; Maxwel Mwase; John Yabe; Tomoyuki Shibahara

    2014-01-01

    This article reports a first case of calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus). Histopathology demonstrated well defined multiple cystic structures containing granular, dark basophilic materials with peripheral granulomatous reaction, characterized by presence of multinucleated giant cells surrounded by a varying amounts of fibrous connective tissues. Special staining with von Kossa revealed black stained deposits confirming the presence of calcium salts.

  12. Unraveling Mill-Rearranged Pedriatic Acute Myeloid Leukemia

    NARCIS (Netherlands)

    B.V. Balgobind (Brian)

    2011-01-01

    textabstractBlood cells are derived from hematopoietic stem cells (HSC) that reside in the bone marrow. HSC’s are multipotent and have the capacity to diff erentiate into the cells of all blood lineages, i.e. erythrocytes, platelets, neutrophils, eosinophils, basophils, monocytes, T and B lymphocyte

  13. Pathology Report for Intraperitoneal Sodium Dichromate Exposure in Rats

    Science.gov (United States)

    2015-08-12

    days, or 14 days before being humanely euthanized under isoflurane anesthesia via exsanguination and organ removal. Post - mortem tissue collection was...Control, Day 1 Post -Injection Rare basophilic tubules, surrounded by minimal lymphocytic infiltrates, are a common background lesion in male Sprague...E-1 Figures 1 Rat #001 Kidney, Vehicle Control, Day 1 Post -Injection

  14. The Subchronic Oral Toxicity of 2,4-Dinitroanisole (DNAN) in Rats

    Science.gov (United States)

    2012-06-01

    only reported compound related microscopic finding was non-specific minimal metaplasia of laryngeal epithelium in rats exposed to 150 mg/m3 (Hoffman...lymphoid, multifocal, minimal. 7. Prostate: Infiltrates, lymphocytic, interstitial and glandular , multifocal, mild. 8. Kidney, left: Basophilic...diffuse, moderate 7. Epididymis: Infiltrates, lymphocytic, interstitial, multifocal, minimal. 8. Prostate: Glandular dilatation, diffuse, moderate

  15. Comprehensive population-based genome sequencing provides insight into hematopoietic regulatory mechanisms

    Science.gov (United States)

    Guo, Michael H.; Nandakumar, Satish K.; Ulirsch, Jacob C.; Zekavat, Seyedeh M.; Buenrostro, Jason D.; Natarajan, Pradeep; Salem, Rany M.; Chiarle, Roberto; Mitt, Mario; Kals, Mart; Pärn, Kalle; Fischer, Krista; Milani, Lili; Mägi, Reedik; Palta, Priit; Gabriel, Stacey B.; Metspalu, Andres; Lander, Eric S.; Kathiresan, Sekar; Hirschhorn, Joel N.; Esko, Tõnu; Sankaran, Vijay G.

    2017-01-01

    Genetic variants affecting hematopoiesis can influence commonly measured blood cell traits. To identify factors that affect hematopoiesis, we performed association studies for blood cell traits in the population-based Estonian Biobank using high-coverage whole-genome sequencing (WGS) in 2,284 samples and SNP genotyping in an additional 14,904 samples. Using up to 7,134 samples with available phenotype data, our analyses identified 17 associations across 14 blood cell traits. Integration of WGS-based fine-mapping and complementary epigenomic datasets provided evidence for causal mechanisms at several loci, including at a previously undiscovered basophil count-associated locus near the master hematopoietic transcription factor CEBPA. The fine-mapped variant at this basophil count association near CEBPA overlapped an enhancer active in common myeloid progenitors and influenced its activity. In situ perturbation of this enhancer by CRISPR/Cas9 mutagenesis in hematopoietic stem and progenitor cells demonstrated that it is necessary for and specifically regulates CEBPA expression during basophil differentiation. We additionally identified basophil count-associated variation at another more pleiotropic myeloid enhancer near GATA2, highlighting regulatory mechanisms for ordered expression of master hematopoietic regulators during lineage specification. Our study illustrates how population-based genetic studies can provide key insights into poorly understood cell differentiation processes of considerable physiologic relevance. PMID:28031487

  16. Measurement of histamine release from human lung tissue ex vivo by microdialysis technique

    DEFF Research Database (Denmark)

    Nissen, Dan; Petersen, Lars Jelstrup; Nolte, H

    1998-01-01

    responses were observed but data could be reproduced within individual donors. Monocyte chemoattractant protein-1, a potent basophil secretagogue, did not induce histamine release in lung tissue which indicated mast cells to be the histamine source. Substance P did not release histamine in the lung tissue...

  17. Significance of Charcot Leyden crystals in hepatic aspirates

    Directory of Open Access Journals (Sweden)

    Misra Vatsala

    2009-01-01

    Full Text Available Charcot Leyden crystals are hexagonal bipyramidal structures localised in the primary granules of the cytoplasm of eosinophils and basophils. Their presence, along with eosinophilic infiltrate, is an indirect evidence of parasitic infestation particularly with Toxocara, Capilliriasis, Ascariasis, or Fasciola. We report here two cases where Charcot Leyden crystals with eosinophilic infiltrate were found in the smears prepared from hepatic abscess.

  18. Human single chain antibodies against heparin: selection, characterization, and effect on coagulation.

    NARCIS (Netherlands)

    Westerlo, E.M.A. van de; Smetsers, T.F.; Dennissen, M.A.B.A.; Linhardt, R.J.; Veerkamp, J.H.; Muijen, G.N.P. van; Kuppevelt, A.H.M.S.M. van

    2002-01-01

    Heparin, located in mast cells and basophilic granulocytes, is widely used as an anticoagulant. It belongs to a class of linear polysaccharides called glycosaminoglycans (GAGs). Using phage display technology, we have selected 19 unique human antiheparin antibodies. Some antibodies react almost excl

  19. Dendritic Cell-Mediated T Cell Proliferation -A Functional Bioindicator of Inflammatory Source-Specific Particulate Matter

    Science.gov (United States)

    Previously we found that dendritic cells (DC) were sensitive functional bioindicators of ambient PM (APM) exposure mediating Th2-allergic inflammation in the draining lymph nodes. Here, the ability of bone-marrow-derived DC (DC) and putative BM-derived basophils (Ba) to present a...

  20. Decitabine, Cytarabine, and Daunorubicin Hydrochloride in Treating Patients With Acute Myeloid Leukemia

    Science.gov (United States)

    2016-07-20

    Adult Acute Basophilic Leukemia; Adult Acute Eosinophilic Leukemia; Adult Acute Monoblastic Leukemia; Adult Acute Monocytic Leukemia; Adult Acute Myeloid Leukemia With Maturation; Adult Acute Myeloid Leukemia With t(9;11)(p22;q23); MLLT3-MLL; Adult Acute Myeloid Leukemia Without Maturation; Adult Acute Myelomonocytic Leukemia; Alkylating Agent-Related Acute Myeloid Leukemia; Secondary Acute Myeloid Leukemia; Untreated Adult Acute Myeloid Leukemia

  1. The in vivo effects of interleukin-3 on histamine levels in non-Hodgkin's lymphoma patients.

    Science.gov (United States)

    Hovgaard, D J; Stahl Skov, P; Nissen, N I

    1997-06-01

    Recombinant human Interleukin-3 (RhIL-3) is a haemopoietic growth factor with effect both on early and differentiated cells, such as eosinophils and basophils, and it also acts as a histamine-releasing agent. The purpose of the present study was to examine whether in vivo rhIL-3 administration after chemotherapy affected basophil histamine levels and whether a concordance between rhIL-3 induced histamine release and side effects during the treatment could be demonstrated. Thirty patients with non-Hodgkin's lymphoma entered the study. All patients received 6 courses of chemotherapy, rhIL-3 was administered subcutaneously once daily after the second and the fourth course of chemotherapy from cycle day 2-15 at the dose levels 0.5, 1.0, 5.0, 7.5 and 10 micrograms/kg with 6 patients at each dose level. In cycle 6 recombinant human Granulocyte-Macrophage Colony-Stimulating Factor (rhGM-CSF) (3.0 micrograms/kg) was administered sequential/concurrent day 9-15 to rhIL-3 (day 2-15) at all dose levels except 7.5 micrograms/kg, where rhIL-3 was given day 2-8 and rhGM-CSF sequential day 9-15. Cycles 1, 3 and 5 served as control cycles with no cytokine therapy. During rhIL-3 treatment, and after CHOP chemotherapy, the basophil counts increased moderately especially during the recovery period day 15-22, and mainly at the two highest dose levels 7.5 and 10 micrograms/kg, but never exceeded the normal upper limit. Histamine levels in basophils were the same in patients before chemotherapy and healthy volunteers, and except from a trend to increased histamine level at 10 micrograms/kg on day 15, no difference was noted between rhIL-3 cycles and control cycles. Within 3-4 hr after rhIL-3 administration, a drop in histamine level in basophils was noted, which could be due to histamine-releasing properties of rhIL-3 as previously demonstrated by in vitro studies. No serious side effects were noted during the cytokine treatment, and despite that most patients had mild flushing of the

  2. A novel human immunoglobulin Fc gamma Fc epsilon bifunctional fusion protein inhibits Fc epsilon RI-mediated degranulation.

    Science.gov (United States)

    Zhu, Daocheng; Kepley, Christopher L; Zhang, Min; Zhang, Ke; Saxon, Andrew

    2002-05-01

    Human mast cells and basophils that express the high-affinity immunoglobulin E (IgE) receptor, Fc epsilon receptor 1 (Fc epsilon RI), have key roles in allergic diseases. Fc epsilon RI cross-linking stimulates the release of allergic mediators. Mast cells and basophils co-express Fc gamma RIIb, a low affinity receptor containing an immunoreceptor tyrosine-based inhibitory motif and whose co-aggregation with Fc epsilon RI can block Fc epsilon RI-mediated reactivity. Here we designed, expressed and tested the human basophil and mast-cell inhibitory function of a novel chimeric fusion protein, whose structure is gamma Hinge-CH gamma 2-CH gamma 3-15aa linker-CH epsilon 2-CH epsilon 3-CH epsilon 4. This Fc gamma Fc epsilon fusion protein was expressed as the predicted 140-kappa D dimer that reacted with anti-human epsilon- and gamma-chain specific antibodies. Fc gamma Fc epsilon bound to both human Fc epsilon RI and Fc gamma RII. It also showed dose- and time-dependent inhibition of antigen-driven IgE-mediated histamine release from fresh human basophils sensitized with IgE directed against NIP (4-hydroxy-3-iodo-5-nitrophenylacetyl). This was associated with altered Syk signaling. The fusion protein also showed increased inhibition of human anti-NP (4-hydroxy-3-nitrophenylacetyl) and anti-dansyl IgE-mediated passive cutaneous anaphylaxis in transgenic mice expressing human Fc epsilon RI alpha. Our results show that this chimeric protein is able to form complexes with both Fc epsilon RI and Fc gamma RII, and inhibit mast-cell and basophil function. This approach, using a Fc gamma Fc epsilon fusion protein to co-aggregate Fc epsilon RI with a receptor containing an immunoreceptor tyrosine-based inhibition motif, has therapeutic potential in IgE- and Fc epsilon RI-mediated diseases.

  3. EAACI taskforce position paper: evidence for autoimmune urticaria and proposal for defining diagnostic criteria.

    Science.gov (United States)

    Konstantinou, G N; Asero, R; Ferrer, M; Knol, E F; Maurer, M; Raap, U; Schmid-Grendelmeier, P; Skov, P S; Grattan, C E H

    2013-01-01

    An autoimmune subset of chronic spontaneous urticaria is increasingly being recognized internationally, based on laboratory and clinical evidence that has accrued over the last 20 years. This evidence has been reviewed by a taskforce of the Dermatology section of the European Academy of Allergy and Clinical Immunology. Functional autoantibodies in chronic urticaria (CU) patient sera have been demonstrated against IgE and FcεRIα by basophil and mast cell histamine release assays and by basophil activation assays. Antibody specificity has been confirmed by immunoassay, but there is a poor correlation between functionality and immunoreactivity. Approximately 25% of CU patients have a positive basophil histamine release assay and show autoreactivity (a positive autologous serum skin test), whereas 50% are negative regarding both. Functionality of CU sera appears to be complement dependent on mast cells but not exclusively on basophils. Basophil activation by CU sera is predominantly restricted to IgG1 and IgG3 subclasses. Circumstantial evidence for CU being an autoimmune disease comes from an observed association with other autoimmune diseases, a strong association between serum functionality and HLA-DR4 haplotype and the good response of CU patients to immunotherapies. It was proposed that a study should be undertaken to prospectively validate potentially relevant clinical criteria (from the history, examination and routinely available clinical investigations) against a new 'gold standard' for the diagnosis of ACU (positive autoreactivity, functional bioassay and immunoassay) to define preliminary criteria sets for the diagnosis of ACU based on clinical and laboratory features with highest individual sensitivity and specificity.

  4. Histamine release-neutralization assay for sera of patients with atopic dermatitis and/or cholinergic urticaria is useful to screen type I hypersensitivity against sweat antigens.

    Science.gov (United States)

    Shindo, Hajime; Ishii, Kaori; Yanase, Yuhki; Suzuki, Hidenori; Hide, Michihiro

    2012-10-01

    We previously reported that about 80 % of patients with atopic dermatitis and 60 % with cholinergic urticaria revealed type I allergy against sweat, by means of skin test against autologous sweat and/or histamine-release test for peripheral blood basophils with semi-purified sweat antigen. In this study, we developed an assay for sera to neutralize histamine-releasing activity of semi-purified sweat antigen. The semi-purified sweat antigen was pre-incubated with serially diluted sera for 30 min at 37 °C and was subjected to histamine-release activity. Histamine release-neutralization (HRN) activities were calculated by measuring the amount of histamine release from basophils in the presence or absence of semi-purified sweat antigen. Of 62 subjects, 39 showed positive histamine release (≥5 %) from their basophils in response to semi-purified sweat antigen, and sera of 34 out of 39 subjects (87.2 %) were also positive in HRN activity (≥10 %). The specificity of the HRN assay was 0.522. Moreover, HRN activities in sera were largely correlated with degrees of histamine release from peripheral blood basophils of the same donors in response to sweat antigen. To identify the substance that neutralizes histamine-release activity, we removed IgE and IgG from the sera of HRN (+) subjects by column chromatography. The HRN activities in 30 out of 42 sera were largely reduced by the removal of IgG. On the other hand, sera of four subjects lost HRN activity by the removal of IgE, suggesting that the majority of HRN (+) subjects have serum IgG against the sweat antigen as well as IgE bound to peripheral basophils. Thus, the HRN assay maybe useful for the screening of type I allergy against sweat antigen.

  5. Purves symposium: 2. Pituitary morphology in relationship to function. General principles.

    Science.gov (United States)

    Purves, H D

    1974-12-25

    It was found that the recognition of the character of a pituitary cell depended on its content of granules. The term "chromophobe" indicated the absence of granules. In the rat pituitary, total thyroxine deficiency resulted in progressive loss of granules from acidophil cells until they became unrecognizable as such. Therefore, counting the cell types as acidophils, basophils, and chromophobes had no significance. However, changes in the apparent proportions of the 3 groups could be used to determine how much thyroxine was needed to reverse changes produced by thyroidectomy. Rats that had been given brassica seed diets developed goiters. In these rats, proliferation of basophil cells occurred. The amount of thyroxine needed to reverse thyroidectomy changes was the same in rats receiving a brassica seed diet as in those on a normal diet. This excluded the possibility of the brassica agent acting as a thyroxine antagonist or of its increasing the thyroxine requirement of the animal. The effect of thyroidectomy was modified by diet. When a diet contained no animal products, the acidophils lost their granules. When acidophil granules were lost, growth ceased. When aciodphil granules had been lost after thyroidectomy, daily doses of thyroxine restored the acidophils and caus ed growth to resume. In animals given potent goitrogens, small doses of thyroxine did not inhibit activation of the thyroid gland, but with larg er doses basophilic changes regressed and the thyroid became inactive. It is concluded that basophil changes were linked with the production of thyrotrophic hormone while acidophil changes which were regressive were related to the failure of growth hormone secretion. When only a single change in hormone production occurs and only a single cell type is modified in the pituitary, that cell type is considered to be the site of production of that particular hormone. By improved technique, it was shown that the basophil cells were of 2 types: "thymotrophs

  6. Acute Effects of Irradiation on People and Animals from Soviet Underground Nuclear Explosions

    Science.gov (United States)

    2008-04-01

    the report, aside from correction of grammatical structure and syntax plus updating radiation units to SI units. Editorial changes were carefully...3.5 1 -6 -eosinophils, % * 3.0 0-5 - basophils , % * 0.5 0- 1 -lymphocytes, % * 28.5 17-40 -monocytes, % * 6.0 2-9 Note: * - index has no sex...1221 3.5 1.8 1.7-5.3 0.3-6.2 0-7.1 Eosinophils, % M 1221 2.7 1.7 1.1-4.4 0.2-5.2 0-6.1 15 Basophils , % M 1221 0.3 0.4 0-0.08 0-1.0 0-1.2

  7. A method for production and determination of histamine releasing activity from human peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Kampen, G T; Poulsen, L K; Reimert, C M

    1997-01-01

    Histamine releasing factors, i.e. cytokines capable of inducing histamine release from basophils or mast cells, have been suggested to be involved in the pathogenesis of, for example, allergic late-phase reactions. Here we describe a controlled method for production and determination of histamine......). The preparations of HRA induced dose- and Ca2+-dependent histamine release from leukocytes. Supernatants of parallel cultures of unstimulated MNC did not induce histamine release. The HRA was neither due to exogenous histamine releasing compounds (e.g. Con A) nor to residual histamine in the preparations of HRA....... The kinetics of HRA induced histamine release (half-maximal release after > 40 min) were slower and more protracted than those of anti-IgE induced histamine release. However, based on a comparison between HRA induced histamine release from leukocytes and purified (97%) basophils, this did not appear to be due...

  8. Novel Functional Aspect of Antihistamines: The Impact of Bepotastine Besilate on Substance P-Induced Events

    Directory of Open Access Journals (Sweden)

    Shun Kitaba

    2009-01-01

    Full Text Available Besides histamine, substance P (SP has been demonstrated to play a crucial role in pruritic skin diseases. Although antihistamines are frequently used for pruritic skin diseases, little is known concerning the effect on an SP-induced event such as mast cell degranulation and the upregulation of adhesion molecules or the nitric oxide (NO synthesis in endothelial cells. Our aim was to study the effect of bepotastine besilate on SP-induced degranulation of rat basophillic leukemia (RBL-2H3 cells and expression of adhesion molecules and NO synthesis in human dermal microvascular endothelial cells (HMVECs. Bepotastine besilate significantly inhibited SP-induced degranulation of RBL-2H3 cells and NO synthesis in HMVECs. Bepotastine besilate significantly inhibited expression of adhesion molecules in HMVESs, while it failed to suppress SP-induced upregulation of the adhesion molecules in HMVECs. Therefore, bepotastine besilate is assumed to act favorably on SP-induced basophil degranulation and NO synthesis in HMVECs.

  9. PERUBAHAN SEBARAN SEL-SEL ASIDOFIL DAN BASOFIL HIPOFISA PENGARUH PEMBERIAN PASAK BUMI (Eurycoma longifolia Jack

    Directory of Open Access Journals (Sweden)

    Hurip Pratomo

    2013-03-01

    Full Text Available Pasak bumi is popular as an aphrodisiac in the traditional medical herb. Boiled drink water of the root powder of pasak bumi is believed can increase the secondary reproduction characters such as libido and the duration of erection. This article is part of the research report that focus on: the distribution of hypophysis acidophil and basophil cells caused of the pasak bumi optimal dose treatment. Research carried out in an experiment using Hemaktosilin Eosin(HE staining method to study micromorphological distribution of the acidophil and basophil cells of the male rat hypophysis after pasak bumi treatment (the dose of 18 mg pasak bumi/200 g body weight during three days, and the control of 1 ml aquadest. Result of the research showed that: Basophyl cells increased significantly in the third days of pasak bumi administered. Meanwhile acidophyl cells decreased in the third days of pasak bumi administered.

  10. An FNA pitfall: Mammary analog secretory carcinoma mistaken for acinic cell carcinoma due to cytoplasmic granules

    Directory of Open Access Journals (Sweden)

    Nouf Hijazi, MD

    2014-12-01

    Full Text Available In the salivary gland, a key differential feature of Mammary analog secretory carcinoma (MASC from acinic cell carcinoma (ACC is the lack of cytoplasmic granules. We report a case of a parotid mass incorrectly diagnosed on fine needle aspirate as acinic cell carcinoma due to many cells with basophilic granules suggesting serous acinar differention. Tumor resection revealed a tumor consistent with low grade adenocarcinoma that had eosinophilic, microvacuolar cytoplasm with distinct basophilic granules staining with PASD and mucicarmine. The diagnosis of MASC was confirmed with stains for GCDF-15, mammoglobin, and S100 and FISH consistent with a t(12;15 translocation. Relying on the absence of cytoplasmic granules as a feature to distinguish ACC from MASC is a diagnostic pitfall.

  11. Acetic orcein staining of prefixed tissue sections.

    Science.gov (United States)

    Reynolds, C; Lillie, R D

    1978-05-01

    Acetic orcein stains formol- and Carnoy-fixed tissues, coloring mast cells, nuclei, basophilic cytoplasm, cerebral corpora amylacea, and cartilage strongly; keratin and erythrocytes moderately; muscle and collagen weakly. Guinea pig Brunner gland and rat colonic goblet cell mucins did not stain. The red nuclear stain contrasts well with the Prussian blue reaction of hemosiderin and the ferric ferricyanide (Turnbull's blue) reaction of enterochromaffin. A weak (0.01%) fast-green FCF stain changes collagen and sometimes smooth muscle to green, without impairing nucleic acid or mast cell staining. Picroindigocarmine gives blue collagen, yellow muscle, and red elastin, nucleic acids and mast cells. Picro-methyl blue tends to override the red nuclear stain. Carnoy fixation is somewhat better for nuclei, formol for basophil cytoplasms.

  12. Induction of histamine release in parasitized individuals by somatic and cuticular antigens from Onchocerca volvulus.

    Science.gov (United States)

    González-Muñoz, M; Gárate, T; Puente, S; Subirats, M; Moneo, I

    1999-06-01

    The host immune response in onchocerciasis is believed to contribute to the clinical manifestations of infection. Mazzotti and chronic inflammatory reactions might be mediated by mechanisms involving specific IgE and reactivity of mast cells and basophils to the parasite antigens. In this report, we show that Onchocerca volvulus antigens are capable of inducing histamine release. Three types of extracts were prepared from the parasite: soluble total, surface, and cuticular collagen. Soluble extracts released histamine in all individuals with onchocerciasis at significantly higher levels (P < 0.05) than those found in endemic controls, but similar levels to those found in patients with mansonellosis. However, cuticular collagen induced significantly (P < 0.01) higher histamine release in patients with onchocerciasis than in those with mansonellosis. No reactivity against human type IV collagen was observed. Implications derived from the presence of sensitized basophils in the pathogenesis of onchocerciasis are discussed.

  13. Two galactose-α-1,3-galactose carrying peptidases from pork kidney mediate anaphylactogenic responses in delayed meat allergy

    DEFF Research Database (Denmark)

    Hilger, Christiane; Fischer, Jörg; Swiontek, Kyra;

    2016-01-01

    BACKGROUND: Serum IgE-antibodies directed at galactose-α-1,3-galactose (α-Gal) are associated with a novel form of delayed anaphylaxis occuring upon consumption of red meat or innards. Pork kidney is known as the most potent trigger of this syndrome, but the culprit allergens have not yet been....... Proteins were identified by peptide mass fingerprinting. Isolated proteins were assayed in ELISA and ELISA inhibition, basophil activation and skin prick test. RESULTS: Several IgE-binding proteins of high molecular weight (100 - >200 kDa) were detected in pork kidney extracts by immunoblot using patient...... resistent to heat denaturation. Pork kidney extract, isolated ACE I and AP-N were able to activate patient basophils and elicit positive responses in skin prick tests. CONCLUSION: Two cell-membrane proteins carrying α-Gal epitopes were identified in pork kidney. For the first time, isolated meat proteins...

  14. Mast cells & Company

    Directory of Open Access Journals (Sweden)

    Friederike eJönsson

    2012-02-01

    Full Text Available Classically, allergy depends on IgE antibodies and on high-affinity IgE receptors expressed by mast cells and basophils. This long accepted IgE/FcεRI/mast cell paradigm, on which the definition of immediate hypersensitivity was based in the Gell and Coomb’s classification, appears too reductionist. Recently accumulated evidence indeed requires that not only IgE but also IgG antibodies, that not only FcεRI but also FcγR of the different types, that not only mast cells and basophils but also neutrophils, monocytes, macrophages, eosinophils, and other myeloid cells by considered as important players in allergy. This view markedly changes our understanding of allergic diseases and, possibly, their treatment.

  15. Immunoglobulin-E reactivity to wine glycoproteins in heavy drinkers

    DEFF Research Database (Denmark)

    Gonzalez-Quintela, Arturo; Gomez-Rial, Jose; Valcarcel, Catalina;

    2011-01-01

    and biological significance of IgE antibodies to N-glycans from wine glycoproteins in heavy drinkers. A structured questionnaire, skin prick tests, serum IgE levels, IgE-immunoblotting to wine extracts, and basophil activation tests were used to characterize 20 heavy drinkers and 10 control subjects. Eleven...... heavy drinkers (55%) showed IgE binding to proteins in wine extracts. The proteins were identified by mass spectrometry as grape-derived vacuolar invertase and thaumatin-like protein. Immunoblot reactivity was closely associated with the presence of IgE to CCDs and was inhibited by preincubation...... with a glycoconjugate containing bromelain-type N-glycans. The same conjugate, CCD-bearing allergens, and wine extracts activated basophils in patients with high-titer CCD-specific IgE but not in healthy controls. There was no relationship between immunoblot reactivity and consumption of any specific type of wine...

  16. [Lead poisoning. A surprising cause of constipation, abdominal pain and anemia].

    Science.gov (United States)

    Hoffmanová, Iva; Kačírková, Petra; Kučerová, Irena; Ševčík, Rudolf; Sánchez, Daniel

    2016-02-01

    This article reports on patient that has been presented with sudden onset of constipation, abdominal pain and normocytic anemia. Gastroscopy and colonoscopy ruled out an organic diseases. In peripheral blood and bone marrow aspirates mears, coarse basophilic stippling of erythrocyte (and erythroblasts) point out a possibility of heavy metal poisoning. The level of plumbemia exceeded 8.4 times the maximal permitted value for common (non-professional) population. A source of poisoning was indentified from a glaze on a ceramic jug, from which the patient had drank tea with lemon for three months. A lead concentration in the tea extract was 227 mg/kg. In developed countries, lead poisoning is a rare diagnosis. As the symptoms are nonspecific, missed diagnoses could occur, especially in sporadic, non-occupational exposure. However, a microscopic evaluation of the peripheral bloods mear with finding of predominantly coarse basophilic stippling of erythrocyte mayle ad to suspicion of lead poisoning.

  17. Journal of Special Operations Medicine. Volume 4, Edition 1, Winter 2004

    Science.gov (United States)

    2004-01-01

    Presenting signs are localized and may include itching, angioedema , or urticaria . These symptoms are caused when mast cells and basophils degranulate...experienced urticaria . One of these had an “allergic reaction” consisting of wheezing, dyspnea, and urticaria --which sounds a lot like anaphylaxis...such as urticaria .2 It was further believed that a nega- tive feedback reaction was associated with the administration of H2 blockers in the setting of

  18. 14-3-3 Proteins, FHA Domains and BRCT Domains in the DNA Damage Response

    OpenAIRE

    Mohammad, Duaa H.; Yaffe, Michael B.

    2009-01-01

    The DNA damage response depends on the concerted activity of protein serine/threonine kinases and modular phosphoserine/threonine binding domains to relay the damage signal and recruit repair proteins. The PIKK family of protein kinases, which includes ATM/ATR/DNA-PK, preferentially phosphorylate Ser-Gln sites, while their basophilic downstream effecter kinases, Chk1/Chk2/MK2 preferentially phosphorylate hydrophobic-X-Arg-X-X-Ser/Thr-hydrophobic sites. A subset of tandem BRCT domains act as p...

  19. AFRRI Reports, First Quarter 1994

    Science.gov (United States)

    1994-06-01

    eosinophil , and basophil numbers, as well as an increase in bone marrow cellularity, GM-CFC, and erythroid burst- forming units (BFU-E) [27]. In... granu - locyte colony-stimulating factor. Blood 78:961 27. Andrews RG, Knitter GH, Bartelmez SH, Langley KE, Far- rar D, Hendren W, Appelbaum FR...cell were found to have degranulated. In addition, endotoxin treatment resulted in the appearance of TNF in the secretory granules of all eosinophils

  20. Food Allergies: The Basics

    OpenAIRE

    Valenta, Rudolf; Hochwallner, Heidrun; Linhart, Birgit; Pahr, Sandra

    2015-01-01

    IgE-associated food allergy affects approximately 3% of the population and has severe effects on the daily life of patients—manifestations occur not only in the gastrointestinal tract but also affect other organ systems. Birth cohort studies have shown that allergic sensitization to food allergens develops early in childhood. Mechanisms of pathogenesis include cross-linking of mast cell– and basophil-bound IgE and immediate release of inflammatory mediators, as well as late-phase and chronic ...

  1. Roles of Macrophages and Neutrophils in the Early Host Response to Bacillus anthracis Spores in a Mouse Model of Infection

    Science.gov (United States)

    2006-01-01

    basophil and lymphocyte populations. In some experiments, control mice were pretreated with saline, and in others, rat IgG (reagent grade, from serum... International . REFERENCES 1. Abalakin, V. A., E. G. Sirina, and T. D. Cherkasova. 1990. The effect of lethal anthrax toxin on the functional activity of...The structure of spores as revealed by mechanical disruption. J. Bacteriol. 66:312–319. 16. Friedlander, A. M. 2000. Anthrax: clinical features

  2. Role of Myelofibrosis in Hematotoxicity of Munition RDX Environmental Degradation Product MNX

    Science.gov (United States)

    2006-09-01

    marrow toxicity including fibrosis. In addition, effects of parent RDX on selected endpoints will be assessed to provide structure -activity information...Fig. 2). Peripheral granulocytes are comprised of neutrophils, basophils , eosinophils and monocytes. Although the CELL-DYNE blood analyzer...colleagues (MacMillan, Denise, et al.) on disposition and metabolism of RDX and MNX is currently undergoing internal review and will be submitted for

  3. Gene Therapy for Fracture Repair

    Science.gov (United States)

    2007-05-01

    structures suggestive of angiogenesis are visible (arrows). (B) Omitting the anti-FGF-2 primary antibody eliminated the immunostaining. 28...Several major families of growth factors, signaling molecules and structural genes are represented, providing one of the most comprehensive surveys...receptor accessory protein NM_012968 IL1 inflammation 1.6 NS 45 IL3 regulated nuclear factor NM_053727 IL3 MHC, eosinphil, basophil stimulation

  4. Histology and Morphology of The Epigonal Organ with Special Referance to the Lymphomyeloid System in Rhinobatos rhinobatos

    OpenAIRE

    Bircan-Yıldırım, Yasemin; Çek, Şehriban; Başusta, Nuri; Atik, Esin

    2011-01-01

    The present study is the first trial to describe the structure of the lymphomyeloid epigonal organ of the Rhinobatos rhinobatos. The morphological structure of the epigonal organ was similar in males and females and it was classified as stingray type. This organ possessed erythrocytes, thrombocytes, lymphocytes and two distinct types of granulocyts (eosinophils and neutrophils). Basophils were not detected in this study. Thrombocytes were variable in shape and lacked visible granules and some...

  5. Intravenous Artesunate: The New Generation of Lifesaving Treatment for Severe Malaria in the Warfighter

    Science.gov (United States)

    2006-11-01

    Artemether Arteether Artesunate Figure 1. Chemical structure of the Artemisinins Artemisinin, extracted from ‘qinghao’ or sweet wormwood...seeking new treatments for malaria, and was first reported in the medical literature in 1979 (QACG, 1979). The structure of artemisinin was...likely related to a temporary arrest of the basophilic erythroblast stage of the developing red blood cells which results in a brief “hole” in the

  6. Necrotizing hepatitis in a domestic pigeon (Columba livia).

    Science.gov (United States)

    Himmel, L; O'Connor, M; Premanandan, C

    2014-11-01

    An adult male domestic pigeon (Columba livia) was presented for necropsy following natural death after a period of chronic weight loss and severe intestinal ascariasis. Histopathologic examination of the liver found moderate to marked, multifocal necrotizing hepatitis with large, basophilic intranuclear inclusion bodies. Transmission electron microscopy of affected hepatocytes demonstrated numerous intra- and perinuclear icosahedral virions arranged in a lattice structure, consistent with adenoviral infection.

  7. Analysis of Long Bone and Vertebral Failure Patterns

    Science.gov (United States)

    1985-02-14

    spinal levels with age. ^ :v ^ In the feline annulus, hexosamine levels were higher in the fe,- I cervical and thoracic annuli compared to the...the annulus are less active in mucopolysaccharide production. In the feline , four cell types have been described in the annulus (Butler and...annulus fibrosus which was eosinophilic peripherally and [v basophilic centrally. Chondrocytes were scattered throughout the annulIIR hetween the

  8. Aspectos morfológicos e citoquímicos dos glóbulos sangüíneos de Caiman crocodilus yacare (Daudin, 1802) (Reptilia, Crocodilia)

    OpenAIRE

    Moura, Weber Leal De [UNIFESP; Matushima, Eliana Reiko; Oliveira, Luiz Waldemar [UNIFESP; EGAMI,MIZUE IMOTO

    1999-01-01

    Samples of peripheral blood were collected from 10 healthy Caiman crocodilus yacare and stained with Leishman, for morphological analysis, or submitted to the following cytochemical methods: PAS, Sudan black B, o-toluidine and mercury bromophenol blue. Seven types of cells were identified: erythrocytes, thrombocytes, heterophils, eosinophils, basophils, lymphocytes and azurofilic monocytes. Erythrocytes were elliptical and positive for bromophenol blue. Some mitotic figures were noted. Spindl...

  9. Calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus

    Directory of Open Access Journals (Sweden)

    Chisoni Mumba

    2014-10-01

    Full Text Available This article reports a first case of calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus. Histopathology demonstrated well defined multiple cystic structures containing granular, dark basophilic materials with peripheral granulomatous reaction, characterized by presence of multinucleated giant cells surrounded by a varying amounts of fibrous connective tissues. Special staining with von Kossa revealed black stained deposits confirming the presence of calcium salts.

  10. Calcinosis circumscripta in a captive African cheetah(Acinonyx jubatus)

    Institute of Scientific and Technical Information of China (English)

    Chisoni; Mumba; David; Squarre; Maxwel; Mwase; John; Yabe; Tomoyuki; Shibahara

    2014-01-01

    This article reports a first case of calcinosis circumscripta in a captive African cheetah(Acinonyx jubatus).Histopathology demonstrated well defined multiple cystic structures containing granular,dark basophilic materials with peripheral granulomatous reaction,characterized by presence of multinucleated giant cells surrounded by a varying amounts of fibrous connective tissues.Special staining with von Kossa revealed black stained deposits confirming the presence of calcium salts.

  11. Perioperative anaphylaxis.

    LENUS (Irish Health Repository)

    Mertes, P M

    2010-07-01

    The incidence of immune-mediated anaphylaxis during anesthesia ranges from 1 in 10,000 to 1 in 20,000. Neuromuscular blocking agents are most frequently incriminated, followed by latex and antibiotics, although any drug or substance used may be a culprit. Diagnosis relies on tryptase measurements at the time of the reaction and skin tests, specific immunoglobulin E, or basophil activation assays. Treatment consists of rapid volume expansion and epinephrine administration titrated to symptom severity.

  12. The C60(FeCp22-Based Cell Proliferation Accelerator

    Directory of Open Access Journals (Sweden)

    Andrei Soldatov

    2013-01-01

    Full Text Available We studied structural and magnetic proprieties of the fulleride C60(FeCp22. The influence of fulleride particles on the cell proliferative activity was also investigated. We found that the proliferative activity of the RINmF5 cells increases (53% versus control in presence of the C60(FeCp22 nanosized particles. Moreover, it was registered that the cell culture became multilayered and secreted basophile matrix.

  13. Omalizumab in Children

    OpenAIRE

    Licari, Amelia; Marseglia, Alessia; Caimmi, Silvia; Castagnoli, Riccardo; Foiadelli, Thomas; Barberi, Salvatore; Marseglia, Gian Luigi

    2014-01-01

    Omalizumab is a recombinant humanized monoclonal antibody that reduces levels of circulating immunoglobulin E (IgE) and expression of IgE high-affinity receptors on mast cells and basophils, interrupting the subsequent allergic inflammatory cascade. Current indications for treatment with omalizumab in pediatric patients are clearly defined and are confined to moderate-to-severe uncontrolled allergic asthma and chronic spontaneous urticaria (CSU). Any other prescription can only be off label. ...

  14. Investigations of the Biologic Activity and Structure Function Relationships of Synthetic Polymers Which Act as Hormones, Behavior Modifiers and Thymic Immunomodulators

    Science.gov (United States)

    1988-07-15

    human peripheral blood basophils . The polyphores produces an energy dependent active release of histamine from both types of cells. The polyphores...demonstrable affinity for calcium, calcium in the medium was required for release of histamine . Each of the hydrophobic copolymers tested had...itr demonstrated that polyphores were ionophores with selectivity for monovalent cations and that they were potent stimulators of histamine release from

  15. Effector Cells and Mechanisms in Chronic Spontaneous Urticaria

    OpenAIRE

    Borzova, Elena

    2014-01-01

    Background: Chronic spontaneous urticaria (CSU) is characterised by weals, angioedema, or both, which occur for six weeks or more. Itchy, red and raised weals in CSU are thought to occur as a result of skin mast cell activation, local vasodilatation and increased vascular permeability which are the cardinal features of the disease. Serum histamine-releasing activity and abnormal basophil function were implicated in the pathophysiology of CSU. We hypothesized that severe and/or persistent CSU ...

  16. Combination treatment of rosuvastatin or atorvastatin, with regular exercise improves arterial wall stiffness in patients with coronary artery disease.

    Directory of Open Access Journals (Sweden)

    Kensuke Toyama

    Full Text Available OBJECTIVE: Statin- and exercise-therapy are both clinically beneficial by preventing cardiovascular events in patients with coronary artery disease (CAD. However, there is no information on the vascular effects of the combination of statins and exercise on arterial wall stiffness in CAD patients. METHODS: The present study is a sub-analysis of PRESET study that determined the effects of 20-week treatment with statins (rosuvastatin, n=14, atorvastatin, n=14 combined with regular exercise on arterial wall stiffness assessed by measurement of brachial and ankle pulse wave velocity (baPWV in CAD patients. RESULTS: The combination of statins and regular exercise significantly improved exercise capacity, lipid profile, including low- and high-density lipoprotein cholesterol, and high-sensitivity C-reactive protein (hs-CRP, baPWV (baseline: 1747 ± 355, at 20 weeks of treatment: 1627 ± 271 cm/s, p=0.008, and basophil count (baseline: 42 ± 32, 20 weeks: 26 ± 15 cells/µL, p=0.007, but had no effect on blood pressure (baseline: 125 ± 22, 20 weeks: 121 ± 16 mmHg. Changes in baPWV correlated significantly with changes in basophil count (r=0.488, p=0.008, but not with age, lipids profile, exercise capacity, or hs-CRP. CONCLUSION: In CAD patients, the combination treatment with statins and exercise resulted in significant amelioration of arterial wall stiffness, at least in part, through reduction of circulating basophils.

  17. cDNA heterogeneity suggests structural variants related to the high-affinity IgE receptor.

    Science.gov (United States)

    Liu, F T; Albrandt, K; Robertson, M W

    1988-08-01

    The high-affinity IgE receptor present on mast cells and basophils is responsible for the IgE-mediated activation of these cells. The current model for this receptor depicts a four-subunit structure, alpha beta gamma 2. A cDNA for the alpha subunit was recently cloned and predicts a structure consisting of two homologous extracellular domains, a transmembrane segment, and a cytoplasmic tail. Using a synthetic oligonucleotide corresponding to the amino-terminal sequence of the alpha subunit, we identified a number of cDNA clones from a rat basophilic leukemia cell cDNA library. Nucleotide sequencing established four different forms of cDNA: one is nearly identical to the published cDNA; the second differs from the first in the 5' untranslated sequence; the other two forms use either one or the other of the 5'-end sequences as above and lack 163 base pairs in the region coding for the second extracellular domain. RNase protection analysis with radioactive RNA probes established the heterogeneity of rat basophilic leukemia cell mRNA with regard to both the 5' and the internal sequences. Our results suggest the existence of at least four different protein forms related to the alpha subunit of the high-affinity IgE receptor.

  18. Haematological values for captive harpy eagle (Harpia harpyja

    Directory of Open Access Journals (Sweden)

    Marcos J. Oliveira

    2014-08-01

    Full Text Available Decreasing of harpy eagle (Harpia harpyja populations in natural environments, mainly in non-preserved areas, makes captive population management an important contribution to genetic diversity conservation. The aim of this study is to evaluate hematological parameters for captive harpy eagles maintained at the wild animals breeding center of Itaipu Binacional, Paraná State, Brazil. Fourteen blood samples from nine harpy eagles were collected from animals of both sexes, of different ages and with no clinical signs of disease. Significant variations were found in haematological values of hematocrit, hemoglobin, mean corpuscular hemoglobin (MCH, mean corpuscular hemoglobin concentration (MCHC, leukocyte, a relative number of heterophils, absolute and relative number of lymphocytes, monocytes, eosinophils, basophils and plasma protein between groups of young (less than six months old and adult birds. Comparing males and females there was variation in the values of erythrocytes, hemoglobin, mean corpuscular volume (MCV, mean corpuscular hemoglobin (MCH on heterophils, absolute and relative number of lymphocytes, eosinophils and basophils. There was also variation in the values of red blood cells, hematocrit, hemoglobin, mean corpuscular volume (MCV, mean corpuscular hemoglobin (MCH, mean corpuscular hemoglobin concentration (MCHC, leukocyte count, absolute number of lymphocytes, eosinophils and basophils among birds that study compared to another reference birds. Due to the limited information available on harpy eagle hematology, this study will be useful to the clinical assessment of birds maintained in captivity.

  19. CHRONIC URTICARIAL IN CHILDHOOD. IMMUNE PATHOLOGY OF CHRONIC URTICARIAL IN CHILDREN (PART I

    Directory of Open Access Journals (Sweden)

    N. A. Sinelnikova

    2013-01-01

    Full Text Available Abstract. This section of our review article deals with prevalence, genetic factors, and some immune mechanisms of chronic urticaria in children. In any form of chronic urticaria, the main pathogenetic role is ascribed to mast cells and basophiles. Increased production of cyto- and chemokines, e.g., CCL2, CХCL8, IL-1, IL-4, IL-6, IL-10, IL-13, IL-17, is observed during chronic inflammatory reactions typical to this disease. Activation of mast cells and basophiles proceeds via specific receptors on the target cell membranes. Autoimmune urticaria is of special importance. In this disorder, autoantibodies are directed against IgE or FCεRIα, thus causing histamine release, due to cross-linking of α-chains of high-affinity IgE receptor, or binding of IgE fixed on the mast cells. Degranulation of basophils is followed by occurrence of activation molecules (СD203с, CD63. Some workers presumet that association between СD203с expression and positive autologous serum skin test may be a sign of autoimmune process. Increased expression of CD40L ligand on the surface of activated T cells suggests enhanced signaling for B cell activation and production of autoantibodies.

  20. Mapping of Lol p I allergenic epitopes by using murine monoclonal antibodies.

    Science.gov (United States)

    Mourad, W; Bernier, D; Jobin, M; Hébert, J

    1989-11-01

    Murine monoclonal antibodies (MAbs) against three non-overlapping epitopes of Lol p I allergen were previously produced and subsequently used for purification of the allergen. In the present study, these MAbs were further characterized, and the biological activity of the purified allergen assessed. The three MAbs were of the IgG isotype and carried a kappa light chain. Their affinity constants were in the range of 7.4-15.1 x 10(-9) mol/l. Purified Lol p I kept its biological activity, as shown by its ability to induce histamine release by basophils of Lol p I-sensitive patients. The profiles of histamine release induced by either Lol p I or crude Lolium perenne extracts were comparable. This observation suggests that human IgE bound to basophils are polyspecific which has been confirmed by immunoblot and inhibition assay. Our data indicated also that Lol p I possesses a major allergenic epitope recognized by all human serum IgE tested. This epitope seems to be partially shared by those recognized by the three MAbs. Finally, preincubation of Lol p I with either one of the Mabs did not affect significantly the basophil-histamine release induced by the purified allergen. This suggests that Lol p I possesses allergenic sites other than the one shared by MAbs and IgE Abs.

  1. Changes in IgE- and Antigen-dependent histamine-release in peripheral blood of Schistosoma mansoni-infected Ugandan fishermen after treatment with praziquantel

    Directory of Open Access Journals (Sweden)

    Kazibwe Francis

    2004-04-01

    Full Text Available Abstract Background Parasite-specific IgE levels correlate with human resistance to reinfection with Schistosoma spp. after chemotherapy. Although the role of eosinophils in schistosomiasis has been the focus of a great deal of important research, the involvement of other Fcε receptor-bearing cells, such as mast cells and basophils, has not been investigated in relation to human immunity to schistosomes. Chemotherapy with praziquantel (PZQ kills schistosomes living in an in vivo blood environment rich in IgE, eosinophils and basophils. This releases parasite Ags that have the potential to cross-link cell-bound IgE. However, systemic hypersensitivity reactions are not induced by treatment. Here, we describe the effects of schistosomiasis, and its treatment, on human basophil function by following changes in total cellular histamine and in vitro histamine-release induced by schistosome Ags or anti-IgE, in blood samples from infected Ugandan fishermen, who are continuously exposed to S. mansoni infection, before and 1-day and 21-days after PZQ treatment. Results There was a significant increase in the total cellular histamine in blood samples at 1-day post-treatment, followed by a very significant further increase by 21-days post-treatment. In vitro histamine-release induced by S. mansoni egg (SEA or worm (SWA Ags or anti-IgE antibody, was significantly reduced 1-day post-treatment. The degree of this reduction correlated with pre-treatment infection intensity. Twenty-1-days post-treatment, SEA-induced histamine-release was still significantly lower than at pretreatment. Histamine-release was not correlated to plasma concentrations of total or parasite-specific IgE, nor to specific IgG4 plasma concentrations. Conclusion The biology of human blood basophils is modulated by S. mansoni infection and praziquantel treatment. Infection intensity-dependent suppression of basophil histamine-release, histamine-dependent resistance to infection, and

  2. 乳头状肾细胞癌的病理及免疫组织化学研究%The Pathological and Immunohistochemical Study of Papillary Renal Cell Carcinoma

    Institute of Scientific and Technical Information of China (English)

    程琦; 宋文刚

    2013-01-01

    Objective To observe the pathological and immunohistochemical study of papillary renal cell carcinoma (PRCC). Methods We analysis 45 cases who had PRCC, all cases were divide into basophilic subtype in 14 cases, acidophilic subtype in 31 cases, according to the organization cell characteristics. Immunohistochemical staining, detection two isoforms epithelial membrane antigen (EMA) and cytokeratin 7 (CK7), CD10, waveform protein (Vim), and two subtypes of Fuhrman classification comparison with the prognosis of patients. Results There had statistical significance of basophilic type and acidophilic type in the EMA, CK7, CD10, Vim expression (P<0.01, P<0.005). Basophilic type and acidophilic type in Fuhrman classification comparison, acidophilic type significantly higher than basophilic type (P<0.005). The 45 patients, follow-up period mortality were 15.6%(7/45), of which type basophil in 2 cases, acidophilic type 5 cases. The rest of the patients were followed up for does not appear during the recurrence or metastases. Conclusion PRCC frequent malignant tumor, its basophilic subtype and acidophilic subtype immune histological characteristics and Fuhrman classification and clinical outcomes are different.%  目的观察乳头状肾细胞癌(PRCC)的病理特点及免疫组织学研究.方法分析PRCC标本共45例,按组织细胞特点嗜碱亚型14例,嗜酸亚型31例.免疫组化染色,检测2种亚型上皮膜抗原(EMA)、细胞角蛋白7(CK7)、CD10、波形蛋白(Vim)及2种亚型Fuhrman分级比较与患者预后分析.结果嗜碱型与嗜酸型在EMA、CK7、CD10、Vim表达方面具有统计学意义(P<0.01,P<0.005).嗜碱型与嗜酸型在Fuhrman分级比较,嗜酸型显著高于嗜碱型,两组分级比较具有统计学意义(P<0.005).本组45例患者,随访期间病死率为15.6%(7/45),其中嗜碱型2例,嗜酸型5例.其余患者随访期间未出现复发或转移.结论乳头状肾细胞癌临床多发恶性肿瘤,其嗜碱亚型和嗜酸亚

  3. Hazelnut (Corylus avellana) vicilin Cor a 11: molecular characterization of a glycoprotein and its allergenic activity.

    Science.gov (United States)

    Lauer, Iris; Foetisch, Kay; Kolarich, Daniel; Ballmer-Weber, Barbara K; Conti, Amedeo; Altmann, Friedrich; Vieths, Stefan; Scheurer, Stephan

    2004-10-15

    In Europe, hazelnuts (Corylus avellana) are a frequent cause of food allergies. Several important hazelnut allergens have been previously identified and characterized. Specific N-glycans are known to induce strong IgE responses of uncertain clinical relevance, but so far the allergenic potential of glycoproteins from hazelnut has not been investigated. The aim of the study was the molecular characterization of the glycosylated vicilin Cor a 11 from hazelnut and the analysis of its allergenic activity. Although MALDI-TOF (matrix-assisted laser-desorption ionization-time-of-flight) MS showed that one of two potential glycosylation sites of Cor a 11 was glycosylated, CD spectroscopy indicated that recombinant and natural Cor a 11 share similar secondary structures. Thus to analyse the impact of the glycan residues of Cor a 11 on IgE binding, the allergenic activity of natural glycosylated Cor a 11 and recombinant Cor a 11 was compared. In addition, the IgE sensitization pattern to recombinant Cor a 11, Cor a 1, Cor a 2 and Cor a 8 of 65 hazelnut allergic patients was determined in vitro. The prevalence of IgE reactivity to hazelnut vicilin Cor a 11 was below 50%. Basophil histamine-release assays were used to determine the allergenic activity of both natural and recombinant Cor a 11 in comparison with Cor a 1, a birch (Betula verrucosa) pollen-related major hazelnut allergen. Both forms of Cor a 11 induced mediator release from basophils to a similar extent, indicating that the hazelnut allergic patients had cross-linking IgE antibodies binding to the protein backbone and not to carbohydrate structures. In comparison to Cor a 1, a 10000-fold higher concentration of Cor a 11 was required to induce similar basophil mediator release. In conclusion, the hazelnut vicilin Cor a 11 is a minor allergen both in regard to prevalence and allergenic potency, whereas its glycan does not contribute to its allergenic activity.

  4. Pathogenic intracellular and autoimmune mechanisms in urticaria and angioedema.

    Science.gov (United States)

    Altman, Katherine; Chang, Christopher

    2013-08-01

    Urticaria and angioedema are common disorders. Chronic urticaria is defined as lasting longer than 6 weeks. Causes of chronic urticaria fall into the following categories: physical, allergic, hereditary, autoimmune, and idiopathic. Basophils and mast cells are the primary effector cells responsible for clinical symptoms and signs. These cells produce and secrete a variety of mediators including histamine, leukotrienes, prostaglandins, cytokines, chemokines, and other pro-inflammatory mediators. This leads to vasodilation, fluid exudation, increased vascular permeability, and accumulation of additional secondary inflammatory cells. Two mechanisms have been investigated as possibly contributing to the pathogenesis of chronic urticaria. One is the development of autoantibodies to FcεRI or IgE on mast cells and basophils. This appears to be responsible for 30-50 % of cases. The other is dysregulation of intracellular signaling pathways involving Syk, SHIP-1, or SHIP-2 in basophils and mast cells. The primary treatment for chronic urticaria is to treat the underlying pathology, if any can be identified. Otherwise, in idiopathic cases, H1 antihistamines, H2 antihistamines, antileukotrienes, and corticosteroids constitute the main pharmacologic treatment modalities. In severe and recalcitrant cases of chronic and autoimmune urticaria, immunosuppressive drugs have been used, most commonly cyclosporin. More recent experimental studies have also suggested that omalizumab, an anti-IgE therapy, may be of benefit. Currently, inhibitors of Syk are also being developed and tested in the laboratory and in animal models. As our understanding of the pathogenesis of idiopathic urticaria increases, development of additional drugs targeting these pathways may provide relief for the significant physical and psychological morbidity experienced by patients with this disorder.

  5. A naturally occurring hypoallergenic variant of vespid Antigen 5 from Polybia scutellaris venom as a candidate for allergen-specific immunotherapy.

    Directory of Open Access Journals (Sweden)

    Sabrina E Vinzón

    Full Text Available Stings by insects from the Hymenoptera order are known to cause life-threatening allergic reactions and impair life quality. Despite the effectiveness of conventional vespid venom immunotherapy, more standardized and safer allergy vaccines are required and recombinant hypoallergenic variants are important clinical tools. Antigen 5 is a major allergen of vespid venoms and it was previously reported that Antigen 5 from Polybia scutellaris (Poly s 5 could be a hypoallergenic variant. In this work we assess the immunological behavior and allergenic activity of Poly s 5 in order to explore its suitability for specific immunotherapy. With this aim, recombinant Poly s 5 was expressed in Pichia pastoris and the presence of cross-reactive epitopes with Pol a 5, a known allergenic Antigen 5, was investigated both at the IgG and IgE levels, by ELISA assays and a basophil-mediator release assay respectively. A molecular model was also built to better understand the relationship between immunological and structural aspects. In mice, Poly s 5 induced IgG antibodies which cross-reacted with Pol a 5. However, Poly s 5 induced only minimal amounts of IgE and was a poor inducer of basophil-mediator release, even when the cells were sensitized with Pol a 5-specific IgE. Moreover, Poly s 5-specific serum showed a specific protective activity and was able to inhibit the Pol a 5-induced basophil degranulation. Structural analysis from the molecular model revealed that a few amino acid substitutions in the N-terminal region of Poly s 5 should lead to an alteration of the surface topography and electrostatic potential of the epitopes which could be responsible for its hypoallergenic behavior. These findings, taken as a whole, show that Poly s 5 is likely a naturally occurring hypoallergenic Antigen 5 variant.

  6. Isoquercitrin suppresses the expression of histamine and pro-inflammatory cytokines by inhibiting the activation of MAP Kinases and NF-κB in human KU812 cells

    Institute of Scientific and Technical Information of China (English)

    LI Li; ZHANG Xiao-Hui; LIU Guang-Rong; LIU Chang; DONG Yin-Mao

    2016-01-01

    Mast cells and basophils are multifunctional effector cells that contain abundant secretory granules in their cytoplasm.Both cell types are involved in a variety of inflammatory and immune events,producing an array of inflammatory mediators,such as cytokines.The aim of the study was to examine whether isoquercitrin modulates allergic and inflammatory reactions in the human basophilic KU812 cells and to elucidate its influence on the phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB activation.The KU812 cells were stimulated with phorbol-12-myristate 13-acetate plus the calcium ionophore A23187 (PMACI).The inhibitory effects of isoquercitrin on the productions of histamine and pro-inflammatory cytokines in the stimulated KU812 cells were measured using cytokine-specific enzyme-linked immunosorbent (ELISA) assays.Western blotting analysis was used to assess the effects of isoquercitrin on the MAPKs and NF-κB protein levels.Our results indicated that the isoquercitrin treatment of PMACI-stimulated KU812 cells significantly reduced the production of histamine and the pro-inflammatory cytokines,such as interleukin (IL)-6,IL-8,IL-1β,and tumor necrosis factor (TNF)-α.The treated cells exhibited decreased phosphorylation of extracellular signal-regulated kinase (ERK),revealing the role of ERK MAPK in isoquercitrin-mediated allergy inhibition.Furthermore,isoquercitrin suppressed the PMACI-mediated activation of NF-κB in the human basophil cells.In conclusion,the results from the present study provide insights into the potential therapeutic use of isoquercitrin for the treatment of inflammatory and allergic reactions.

  7. A naturally occurring hypoallergenic variant of vespid Antigen 5 from Polybia scutellaris venom as a candidate for allergen-specific immunotherapy.

    Science.gov (United States)

    Vinzón, Sabrina E; Marino-Buslje, Cristina; Rivera, Elena; Biscoglio de Jiménez Bonino, Mirtha

    2012-01-01

    Stings by insects from the Hymenoptera order are known to cause life-threatening allergic reactions and impair life quality. Despite the effectiveness of conventional vespid venom immunotherapy, more standardized and safer allergy vaccines are required and recombinant hypoallergenic variants are important clinical tools. Antigen 5 is a major allergen of vespid venoms and it was previously reported that Antigen 5 from Polybia scutellaris (Poly s 5) could be a hypoallergenic variant. In this work we assess the immunological behavior and allergenic activity of Poly s 5 in order to explore its suitability for specific immunotherapy. With this aim, recombinant Poly s 5 was expressed in Pichia pastoris and the presence of cross-reactive epitopes with Pol a 5, a known allergenic Antigen 5, was investigated both at the IgG and IgE levels, by ELISA assays and a basophil-mediator release assay respectively. A molecular model was also built to better understand the relationship between immunological and structural aspects. In mice, Poly s 5 induced IgG antibodies which cross-reacted with Pol a 5. However, Poly s 5 induced only minimal amounts of IgE and was a poor inducer of basophil-mediator release, even when the cells were sensitized with Pol a 5-specific IgE. Moreover, Poly s 5-specific serum showed a specific protective activity and was able to inhibit the Pol a 5-induced basophil degranulation. Structural analysis from the molecular model revealed that a few amino acid substitutions in the N-terminal region of Poly s 5 should lead to an alteration of the surface topography and electrostatic potential of the epitopes which could be responsible for its hypoallergenic behavior. These findings, taken as a whole, show that Poly s 5 is likely a naturally occurring hypoallergenic Antigen 5 variant.

  8. Effect of seasonal variation on adult clinical laboratory parameters in Rwanda, Zambia, and Uganda: implications for HIV biomedical prevention trials.

    Directory of Open Access Journals (Sweden)

    Eugene Ruzagira

    Full Text Available To investigate the effect of seasonal variation on adult clinical laboratory parameters in Rwanda, Zambia, and Uganda and determine its implications for HIV prevention and other clinical trials.Volunteers in a cross-sectional study to establish laboratory reference intervals were asked to return for a seasonal visit after the local season had changed from dry to rainy or vice versa. Volunteers had to be clinically healthy, not pregnant and negative for HIV, Hepatitis B and C, and syphilis infection at both visits. At each visit, blood was taken for measurement of hemoglobin, haematocrit, mean corpuscular volume, red blood cells, platelets, total white blood cells (WBC, neutrophils, lymphocytes, monocytes, eosinophils, basophils, CD4/CD8 T cells, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, direct bilirubin, total bilirubin, total immunoglobulin gamma, total protein, creatinine, total amylase, creatine phosphokinase and lactate dehydrogenase (LDH. Consensus dry season reference intervals were applied to rainy season values (and vice versa and the proportion of 'out-of-range' values determined. Percentage differences between dry and rainy season parameter mean values were estimated.In this cohort of 903 volunteers, less than 10.0% of consensus parameter (except LDH values in one season were "out-of-range" in the other. Twenty-two (22 percent of rainy season LDH values fell outside of the consensus dry season interval with the higher values observed in the rainy season. Variability between consensus seasonal means ranged from 0.0% (total WBC, neutrophils, monocytes, basophils, and direct bilirubin to 40.0% (eosinophils. Within sites, the largest seasonal variations were observed for monocytes (Masaka, 11.5%, LDH (Lusaka, 21.7%, and basophils (Kigali, 22.2%.Seasonality had minimal impact on adult clinical laboratory parameter values in Rwanda, Zambia, and Uganda. Seasonal variation may not be an important factor in the

  9. Identification of autoclave-resistant Anisakis simplex allergens.

    Science.gov (United States)

    Carballeda-Sangiao, Noelia; Olivares, Fabiola; Rodriguez-Mahillo, Ana I; Careche, Mercedes; Tejada, Margarita; Moneo, Ignacio; González-Muñoz, Miguel

    2014-04-01

    Anisakis simplex is a fish parasite able to induce allergic reactions in humans infected when eating raw or undercooked fish parasitized with viable third-stage larvae. Some authors claim that exposure to nonviable Anisakis material can result in allergic symptoms in previously sensitized patients, indicating that parasite allergens are resistant to the thermal treatments of usual cooking procedures. Furthermore, some patients report symptoms after eating canned fish. The aim of this work was the analysis of parasite allergen stability in heating to 121 °C in an autoclave to simulate the thermal process applied to canned fish. Third-stage larvae were subjected to autoclaving for 20, 40, and 80 min, and parasite crude extracts were analyzed by electrophoresis, immunoblotting, and a flow-cytometric basophil activation test. Allergens resistant to autoclaving were separated by reversed-phase high-performance liquid chromatography and identified by ion trap mass spectrometry. Protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that autoclaving considerably reduced the number and intensity of identifiable protein bands in a time-dependent manner. Several allergens were detected by immunoblotting with a pool of A. simplex allergic patients' sera after autoclaving. Allergens of 9 and 14 kDa resistant to autoclaving were identified as Ani s 4 and Ani s 1 allergens, respectively. Functional analysis showed that allergens retain their capacity to activate basophils even after autoclaving for 80 min. In conclusion, some relevant A. simplex allergens retain their capacity to bind immunoglobulin E and activate basophils after being subjected to autoclaving, which is a method equivalent to that used in industrial canning processes.

  10. Human IgE-independent systemic anaphylaxis.

    Science.gov (United States)

    Finkelman, Fred D; Khodoun, Marat V; Strait, Richard

    2016-06-01

    Anaphylaxis is a rapidly developing, life-threatening, generalized or systemic allergic reaction that is classically elicited by antigen crosslinking of antigen-specific IgE bound to the high-affinity IgE receptor FcεRI on mast cells and basophils. This initiates signals that induce cellular degranulation with release and secretion of vasoactive mediators, enzymes, and cytokines. However, IgE-independent mechanisms of anaphylaxis have been clearly demonstrated in experimental animals. These include IgG-dependent anaphylaxis, which involves the triggering of mediator release by IgG/antigen complex crosslinking of FcγRs on macrophages, basophils, and neutrophils; anaphylaxis mediated by binding of the complement-derived peptides C3a and C5a to their receptors on mast cells, basophils, and other myeloid cells; and direct activation of mast cells by drugs that interact with receptors on these cells. Here we review the mechanisms involved in these IgE-independent forms of anaphylaxis and the clinical evidence for their human relevance. We conclude that this evidence supports the existence of all 3 IgE-independent mechanisms as important causes of human disease, although practical and ethical considerations preclude their demonstration to the degree of certainty possible with animal models. Furthermore, we cite evidence that different clinical situations can suggest different mechanisms as having a primal role in anaphylaxis and that IgE-dependent and distinct IgE-independent mechanisms can act together to increase anaphylaxis severity. As specific agents become available that can interfere with mechanisms involved in the different types of anaphylaxis, recognition of specific types of anaphylaxis is likely to become important for optimal prophylaxis and therapy.

  11. A disease marker for aspirin-induced chronic urticaria.

    Science.gov (United States)

    Hsieh, Chia-Wei; Lee, Jeen-Wei; Liao, En-Chih; Tsai, Jaw-Ji

    2014-07-15

    There are currently no diagnostic methods in vitro for aspirin-induced chronic urticaria (AICU) except for the provocation test in vivo. To identify disease markers for AICU, we investigated the single nucleotide polymorphism (SNP) of the promoter loci of high-affinity IgE receptor (FcεRIα) and CD203c expression level in Chinese patients with AICU. We studied two genotypic and allelic frequencies of rs2427827 (-344C/T) and rs2251746 (-66T/C) gene polymorphisms of FcεRIα in 20 patients with AICU, 52 subjects with airway hypersensitivity without aspirin intolerance, and 50 controls in a Chinese population. The results showed that the frequencies of two SNPs (-344C>T, -66C>T) were similar to the normal controls. The allele frequency of -344CC was significantly higher in the patients with AICU compared to those with airway sensitivity (p=0.019). We also studied both histamine release and CD203c expression on KU812 cells to assess aspirin-induced basophil activation. We found that the activity of basophil activation of AICU was significantly higher in the patients with AICU compared to those with airway hypersensitivity without aspirin intolerance. The mean fluorescence intensity of the CD203c expression were 122.5±5.2 vs. 103.3±3.3 respectively, (phistamine release were 31.3%±7.4% vs. -24.0%±17.5%, (phistamine release were significantly up-regulated by aspirin, they were not affected by anti-IgE antibodies. These results suggest that a single SNP of FcεRIα (-344C>T) is less likely to develop AICU and the basophil activation activity in the sera by measuring CD203c expression can be applicable to confirm the diagnosis of AICU.

  12. Comparison of the in vivo autologous skin test with in vitro diagnostic tests for diagnosis of chronic autoimmune urticaria.

    Science.gov (United States)

    Altrich, Michelle L; Halsey, John F; Altman, Leonard C

    2009-01-01

    Previous studies indicate that 30-50% of chronic urticaria patients have an autoimmune etiology. Clinical diagnosis of autoimmune urticaria is supported with the autologous serum skin test. The purpose of this study was to compare two laboratory tests for measurement of IgG autoantibodies to IgE or IgE receptors and compare the results with the autologous serum and plasma skin tests. We performed skin tests and two functional in vitro tests, basophil histamine release, and CD63 up-regulation to detect autoantibodies relevant to autoimmune urticaria. Both sera and citrated plasma were evaluated in the autologous skin test and histamine release assay. Thyroid autoantibodies were also measured. Basophils were incubated with patient plasma, sera, buffer, or anti-IgE. The cells were analyzed for CD63 expression and the supernatants were recovered for histamine analysis. There was high correlation between CD63 up-regulation and histamine release assays, but histamine release was more sensitive. There was a high concordance between sera and citrated plasma for the skin test. Sera from chronic urticaria patients produced higher mean histamine release (23%) compared with citrated plasma (12%). Thirty-one percent of patients positive in the histamine release assay were also positive for thyroid autoantibodies. This compares with 12% who were negative in the histamine release assay. These data show that in vitro basophil histamine release can be used to measure antibodies to FceRI, FceRII/CD23, or IgE and identify patients with autoimmune urticaria.

  13. Isoquercitrin suppresses the expression of histamine and pro-inflammatory cytokines by inhibiting the activation of MAP Kinases and NF-κB in human KU812 cells.

    Science.gov (United States)

    Li, Li; Zhang, Xiao-Hui; Liu, Guang-Rong; Liu, Chang; Dong, Yin-Mao

    2016-06-01

    Mast cells and basophils are multifunctional effector cells that contain abundant secretory granules in their cytoplasm. Both cell types are involved in a variety of inflammatory and immune events, producing an array of inflammatory mediators, such as cytokines. The aim of the study was to examine whether isoquercitrin modulates allergic and inflammatory reactions in the human basophilic KU812 cells and to elucidate its influence on the phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB activation. The KU812 cells were stimulated with phorbol-12-myristate 13-acetate plus the calcium ionophore A23187 (PMACI). The inhibitory effects of isoquercitrin on the productions of histamine and pro-inflammatory cytokines in the stimulated KU812 cells were measured using cytokine-specific enzyme-linked immunosorbent (ELISA) assays. Western blotting analysis was used to assess the effects of isoquercitrin on the MAPKs and NF-κB protein levels. Our results indicated that the isoquercitrin treatment of PMACI-stimulated KU812 cells significantly reduced the production of histamine and the pro-inflammatory cytokines, such as interleukin (IL)-6, IL-8, IL-1β, and tumor necrosis factor (TNF)-α. The treated cells exhibited decreased phosphorylation of extracellular signal-regulated kinase (ERK), revealing the role of ERK MAPK in isoquercitrin-mediated allergy inhibition. Furthermore, isoquercitrin suppressed the PMACI-mediated activation of NF-κB in the human basophil cells. In conclusion, the results from the present study provide insights into the potential therapeutic use of isoquercitrin for the treatment of inflammatory and allergic reactions.

  14. Release of Bet v 1 from birch pollen from 5 European countries. Results from the HIALINE study

    Science.gov (United States)

    The HIALINE working Group; Buters, Jeroen T. M.; Thibaudon, Michel; Smith, Matt; Kennedy, Roy; Rantio-Lehtimäki, Auli; Albertini, Roberto; Reese, Gerald; Weber, Bernhard; Galan, Carmen; Brandao, Rui; Antunes, Celia M.; Jäger, Siegfried; Berger, Uwe; Celenk, Sevcan; Grewling, Łukasz; Jackowiak, Bogdan; Sauliene, Ingrida; Weichenmeier, Ingrid; Pusch, Gudrun; Sarioglu, Hakan; Ueffing, Marius; Behrendt, Heidrun; Prank, Marje; Sofiev, Mikhail; Cecchi, Lorenzo

    2012-08-01

    Exposure to allergens is pivotal in determining sensitization and allergic symptoms in individuals. Pollen grain counts in ambient air have traditionally been assessed to estimate airborne allergen exposure. However, the exact allergen content of ambient air is unknown. We therefore monitored atmospheric concentrations of birch pollen grains and the matched major birch pollen allergen Bet v 1 simultaneously across Europe within the EU-funded project HIALINE (Health Impacts of Airborne Allergen Information Network).Pollen count was assessed with Hirst type pollen traps at 10 l min-1 at sites in France, United Kingdom, Germany, Italy and Finland. Allergen concentrations in ambient air were sampled at 800 l min-1 with a Chemvol® high-volume cascade impactor equipped with stages PM > 10 μm, 10 μm > PM > 2.5 μm, and in Germany also 2.5 μm > PM > 0.12 μm. The major birch pollen allergen Bet v 1 was determined with an allergen specific ELISA. Bet v 1 isoform patterns were analyzed by 2D-SDS-PAGE blots and mass spectrometric identification. Basophil activation was tested in an FcɛR1-humanized rat basophil cell line passively sensitized with serum of a birch pollen symptomatic patient.Compared to 10 previous years, 2009 was a representative birch pollen season for all stations. About 90% of the allergen was found in the PM > 10 μm fraction at all stations. Bet v 1 isoforms pattern did not vary substantially neither during ripening of pollen nor between different geographical locations. The average European allergen release from birch pollen was 3.2 pg Bet v 1/pollen and did not vary much between the European countries. However, in all countries a >10-fold difference in daily allergen release per pollen was measured which could be explained by long-range transport of pollen with a deviating allergen release. Basophil activation by ambient air extracts correlated better with airborne allergen than with pollen concentration.Although Bet v 1 is a mixture of different

  15. Navigating the Updated Anaphylaxis Parameters

    Directory of Open Access Journals (Sweden)

    Kemp Stephen F

    2007-06-01

    Full Text Available Anaphylaxis, an acute and potentially lethal multi-system clinical syndrome resulting from the sudden, systemic degranulation of mast cells and basophils, occurs in a variety of clinical scenarios and is almost unavoidable inmedical practice. Healthcare professionalsmust be able to recognize its features, treat an episode promptly and appropriately, and be able to provide recommendations to prevent future episodes. Epinephrine, administered immediately, is the drug of choice for acute anaphylaxis. The discussion provides an overview of one set of evidence-based and consensus parameters for the diagnosis and management of anaphylaxis.

  16. Innate-Type and Acquired-Type Allergy Regulated by IL-33

    Directory of Open Access Journals (Sweden)

    Tomohiro Yoshimoto

    2014-01-01

    Full Text Available We propose two types of allergic response: IgE-dependent and IgE-independent, and designate these as 'acquired-type allergy' and 'innate-type allergy', respectively. IL-33 stimulates both innate (basophils, mast cells, or group 2 innate lymphoid cells and acquired (Th2 cells allergy-related cells to induce and/or augment Th2 cytokine production, which leads to eosinophilic inflammation in vivo. Thus, IL-33 is an essential regulator for both 'innate-type allergy' and 'acquired-type allergy', and might be an attractive therapeutic target for allergic diseases.

  17. Mammalian Toxicity of Munition Compounds. Phase II. Effects of Multiple Doses. Part III. 2,6-Dinitrotoluene

    Science.gov (United States)

    1976-07-01

    nucleated erythrocytes and granu - locytic cells. Since this was seen in dogs given this dose for 13 weeks (Table 16), and in dogs given Ligher doses, it was...in the testes of the male controls (Tables 48 and 50), minimal microcalculi and focal mononuclear infiltration in the kidney, eosinophil infiltration... Eosinophils (%) 3.2 ± 0.8 2 - 4 Basophils (%) 0 * 0 0 - 0 Monocytes (%) 3.4 ± 0.9 3 - 5 Atypical (%) 0*0 0 - 0 Nucleated RBC (%) 0 0 0 - 0I Methemoglobin

  18. [Calcium polystyrene sulfonate induced colonic necrosis in patient with chronic kidney disease].

    Science.gov (United States)

    Lee, Sung Hoa; Kim, Sung Jung; Kim, Go Eun; Lee, Woo Jin; Hong, Won Ki; Baik, Gwang Ho; Choi, Young Hee; Kim, Dong Joon

    2010-04-01

    A 63-year-old woman was admitted due to right upper quadrant abdominal pain. She was going through hemodialysis due to end stage renal disease and taking calcium polystyrene sulfonate orally and rectally due to hyperkalemia. Colonoscopy showed a circular ulcerative mass on the proximal ascending colon. Biopsy specimen from the mass showed inflammation and necrotic debris. It also revealed basophilic angulated crystals which were adherent to the ulcer bed and normal mucosa. These crystals were morphologically consistent with calcium polystyrene sulfonate. She was diagnosed with calcium polystyrene phosphate induced colonic necrosis and improved with conservative treatment.

  19. The epidemiology of anaphylaxis in Europe

    DEFF Research Database (Denmark)

    Panesar, S S; Javad, S; de Silva, D;

    2013-01-01

    BACKGROUND: Anaphylaxis is an acute, potentially fatal, multi-organ system, allergic reaction caused by the release of chemical mediators from mast cells and basophils. Uncertainty exists around epidemiological measures of incidence and prevalence, risk factors, risk of recurrence, and death due...... and analytical epidemiological investigations, and studies involving analysis of routine data. RESULTS: Our searches identified a total of 5,843 potentially eligible studies, of which 49 satisfied our inclusion criteria. Of these, three were suitable for pooled estimates of prevalence. The incidence rates...

  20. cDNA heterogeneity suggests structural variants related to the high-affinity IgE receptor.

    OpenAIRE

    Liu, F T; Albrandt, K; Robertson, M W

    1988-01-01

    The high-affinity IgE receptor present on mast cells and basophils is responsible for the IgE-mediated activation of these cells. The current model for this receptor depicts a four-subunit structure, alpha beta gamma 2. A cDNA for the alpha subunit was recently cloned and predicts a structure consisting of two homologous extracellular domains, a transmembrane segment, and a cytoplasmic tail. Using a synthetic oligonucleotide corresponding to the amino-terminal sequence of the alpha subunit, w...

  1. Immunotherapy with Allergen Peptides

    Directory of Open Access Journals (Sweden)

    Larché Mark

    2007-06-01

    Full Text Available Specific allergen immunotherapy (SIT is disease-modifying and efficacious. However, the use of whole allergen preparations is associated with frequent allergic adverse events during treatment. Many novel approaches are being designed to reduce the allergenicity of immunotherapy preparations whilst maintaining immunogenicity. One approach is the use of short synthetic peptides which representing dominant T cell epitopes of the allergen. Short peptides exhibit markedly reduced capacity to cross link IgE and activate mast cells and basophils, due to lack of tertiary structure. Murine pre-clinical studies have established the feasibility of this approach and clinical studies are currently in progress in both allergic and autoimmune diseases.

  2. Epitope grafting, re-creating a conformational Bet v 1 antibody epitope on the surface of the homologous apple allergen Mal d 1

    DEFF Research Database (Denmark)

    Holm, Jens; Ferreras, Mercedes; Ipsen, Henrik;

    2011-01-01

    Birch-allergic patients often experience oral allergy syndrome upon ingestion of vegetables and fruits, most prominently apple, that is caused by antibody cross-reactivity of the IgE antibodies in patients to proteins sharing molecular surface structures with the major birch pollen group 1 allergen...... scaffold molecule without loss of epitope functionality. Furthermore, we show that increasing surface similarity to Bet v 1 of Mal d 1 variants by substitution of 6-8 residues increased the ability to trigger basophil histamine release with blood from birch-allergic patients not responding to natural Mal d...

  3. Regulation of the Il4 Gene Is Independently Controlled by Proximal and Distal 3′ Enhancers in Mast Cells and Basophils▿

    OpenAIRE

    Yagi, Ryouji; Tanaka, Shinya; Motomura, Yasutaka; Kubo, Masato

    2007-01-01

    Mast cells and basophils are known to be a critical interleukin 4 (IL-4) source for establishing Th2 protective responses to parasitic infections. Chromatin structure and histone modification patterns in the Il13/Il4 locus of mast cells were similar to those of IL-4-producing type 2 helper T cells. However, using a transgenic approach, we found that Il4 gene expression was distinctly regulated by individual cis regulatory elements in cell types of different lineages. The distal 3′ element con...

  4. Unlocking the allergenic structure of the major house dust mite allergen der f 2 by elimination of key intramolecular interactions.

    Science.gov (United States)

    Takai, T; Ichikawa, S; Yokota, T; Hatanaka, H; Inagaki, F; Okumura, Y

    2000-11-03

    We report on the structural background of the remarkable reduction of allergenicity in engineering of the major house dust mite allergen Der f 2. Disruption of intramolecular disulfide bonds in Der f 2 caused extensive conformational change that was monitored by circular dichroism and gel-filtration analysis. The degree of conformational change correlated well with the degree of reductions in the capacity to bind IgE and to induce histamine release from basophils in mite-allergic patients. Loosening the rigid tertiary structure by elimination of key intramolecular interactions is an effective strategy to reduce the number of high affinity IgE epitopes of allergen vaccine.

  5. Effects of deuterium oxide on cell growth and vesicle speed in RBL-2H3 cells

    Directory of Open Access Journals (Sweden)

    Roshni S. Kalkur

    2014-09-01

    Full Text Available For the first time we show the effects of deuterium oxide on cell growth and vesicle transport in rat basophilic leukemia (RBL-2H3 cells. RBL-2H3 cells cultured with 15 moles/L deuterium showed decreased cell growth which was attributed to cells not doubling their DNA content. Experimental observations also showed an increase in vesicle speed for cells cultured in deuterium oxide. This increase in vesicle speed was not observed in deuterium oxide cultures treated with a microtubule-destabilizing drug, suggesting that deuterium oxide affects microtubule-dependent vesicle transport.

  6. Allergic Rhinitis: Mechanisms and Treatment.

    Science.gov (United States)

    Bernstein, David I; Schwartz, Gene; Bernstein, Jonathan A

    2016-05-01

    The prevalence of allergic rhinitis (AR) has been estimated at 10% to 40%, and its economic burden is substantial. AR patients develop specific immunoglobulin E (IgE) antibody responses to indoor and outdoor environmental allergens with exposure over time. These specific IgE antibodies bind to high-affinity IgE receptors on mast cells and basophils. Key outcome measures of therapeutic interventions include rhinitis symptom control, rescue medication requirements, and quality-of-life measures. A comprehensive multiple modality treatment plan customized to the individual patient can optimize outcomes.

  7. AFRRI Reports Third Quarter, July - September 1991

    Science.gov (United States)

    1991-10-01

    learned ,ine pulmonar arter endothelial cells (251), and rat about the physiology of phagocytic cells, including the basophilic leukemia cells (154). role...m. nge r get r~atiun I n t ht. prtuni I-onU titteci I r I IL-uu.J. channel anasl. of thle potasmom permeailikt3 in lIeLa cancer Thi. (lien, 265... pulmonar . macrophages. 2V7. SEGEL. G. M. AND M. A. LICIITMAN. Potassium transport in Xiernry 21. 6321-6.26. 19F2. hoinam bliood .mphtoe 71 Lreauieaa tilt ph

  8. Combination Chemotherapy in Treating Young Patients With Down Syndrome and Acute Myeloid Leukemia or Myelodysplastic Syndromes

    Science.gov (United States)

    2017-02-07

    Childhood Acute Basophilic Leukemia; Childhood Acute Eosinophilic Leukemia; Childhood Acute Erythroleukemia (M6); Childhood Acute Megakaryocytic Leukemia (M7); Childhood Acute Minimally Differentiated Myeloid Leukemia (M0); Childhood Acute Monoblastic Leukemia (M5a); Childhood Acute Monocytic Leukemia (M5b); Childhood Acute Myeloblastic Leukemia With Maturation (M2); Childhood Acute Myeloblastic Leukemia Without Maturation (M1); Childhood Acute Myelomonocytic Leukemia (M4); Childhood Myelodysplastic Syndromes; de Novo Myelodysplastic Syndromes; Secondary Acute Myeloid Leukemia; Secondary Myelodysplastic Syndromes; Untreated Childhood Acute Myeloid Leukemia and Other Myeloid Malignancies

  9. Bryophyte flora of the Uvac river gorge (Southwest Serbia

    Directory of Open Access Journals (Sweden)

    Veljić M.

    2006-01-01

    Full Text Available In the examined area, 165 taxa were found and identified: 139 taxa from the class Bryopsida and 26 taxa from the class Marchantiopsida. Nine species are red-listed in Serbia. Material was collected from 62 localities, which were analyzed for similarity of chorological and ecological features using the Jaccard similarity index. Analysis of floristic elements and phytogeographic distribution showed that the greatest number of taxa are temperate elements with Holarctic distribution. Results of ecological analysis showed that in regard to the substratum aspect, terricolous, basophilous, and indifferent species were dominant. In relation to the ecological parameter humidity, most species were mesophilous. The majority of identified bryophytes were sciophilous taxa. .

  10. The effects of vitamin A supplementation with measles vaccine on leucocyte counts and in vitro cytokine production

    DEFF Research Database (Denmark)

    Jensen, Kristoffer Jarlov; Fisker, Ane Bærent; Andersen, Andreas;

    2016-01-01

    on overall mortality of providing VAS with vaccines in Guinea-Bissau, we conducted an immunological sub-study of VAS v. placebo with MV, analysing leucocyte counts, whole blood in vitro cytokine production, vitamin A status and concentration of C-reactive protein (CRP). VAS compared with placebo...... was associated with an increased frequency of CRP≥5 mg/l (28 v. 12 %; P=0·005). Six weeks after supplementation, VAS had significant sex-differential effects on leucocyte, lymphocyte, monocyte and basophil cell counts, decreasing them in males but increasing them in females. Mainly in females, the effect of VAS...

  11. Macroscopic and microscopic aspects of collared peccary and white-lipped peccary placenta

    DEFF Research Database (Denmark)

    Santos, T.C.; Dantzer, Vibeke; Jones, C.J.P.

    2006-01-01

    a placental barrier 3 µm or less thick. The columnar uterine glandular epithelium has a subpopulation of granules staining with Perl's Prussion blue reaction, suggesting iron secretion. In areolar areas, the trophoblast cells show apical microvilli, a basophilic cytoplasm with electron-dense intracellular...... placentae show no trophoblast invasion into the uterine epithelium and there is interdigitation between fetal and maternal microvilli. Two distinct regions of the fetomaternal interface can be identified: the interareolar and the areolar regions. The uterine epithelium has eosinophilic cytoplasm...

  12. Crystallization and preliminary X-ray analysis of birch-pollen allergen Bet v 1 in complex with a murine monoclonal IgG Fab' fragment

    DEFF Research Database (Denmark)

    Spangfort, M D; Mirza, Osman Asghar; Gajhede, M

    1999-01-01

    The human type I allergic response is characterized by the presence of allergen-specific serum immunoglobulin E (IgE). Allergen-mediated cross-linking of receptor-bound IgE on the surface of mast cells and circulating basophils triggers the release of mediators, resulting in the development of th......, with unit-cell parameters a = 91.65, b = 99.14, c = 108.90 A, alpha = 105.7, beta = 98.32, gamma = 97.62 degrees, and diffract to 2.9 A resolution when analyzed at 100 K using synchrotron-generated X--rays....

  13. Murine B cells regulate serum IgE levels in a CD23-dependent manner1

    OpenAIRE

    2010-01-01

    The manifestations of allergic disorders are closely tied to the biologic effects of IgE activation with antigen. In immediate hypersensitivity reactions, IgE effector function requires prior binding to innate immune cells, primarily mast cells and basophils, with the blood acting as a reservoir for unbound IgE. As the severity of allergic disease is proportional to the size of this unbound IgE pool, we hypothesized that cellular mechanisms exist to limit the size and/or enhance the clearance...

  14. Studies on the Tolerating Mechanism for Sulfide in Urechis unicinctus ( Echiura : Urechidae) ——Cytological Observation on Urechis unicinctus in Different Hydrogen Sulfide Environment

    Institute of Scientific and Technical Information of China (English)

    张志峰; 邵明瑜; 康庆浩; 金在敏

    2003-01-01

    This study on the cytological changes of the body wall, aspiratory intestine and crissalbursa in Urechis unicinctus by light microscope (LM) and transmission electron microscope (TEM)showed that the difference between the body wall and the natural environment rich in H2 S was notobvious; that the wall color of the aspiratory intestine in H2 S rich environment changed from normalsemitransparency to dark brown; that its epithelia were disassembled and the electron density of itscytoplasm matrix was lower; and that in H2 S rich enviromnent many basophilic granules occurred inthe epithelia of the crissal bursa. Granules with single membrane and myelinefingure were foundwith TEM.

  15. 166 Assessment of Chronic Spontaneous Urticaria by Serum-Induced TNF & ALPHA; and MMP-9 Release

    OpenAIRE

    Falkencronec, Sidsel; Poulsen, Lars; Maurer, Marcus; Bindslev-Jensen, Carsten; Skov, Per Stahl

    2012-01-01

    Background Previous studies from our group have demonstrated that IgE-mediated basophil activation leads to release of TNFα that in turn can induce matrix metallo-proteinase-9 (MMP-9) release from monocytes. We wished to investigate if serum from chronic spontaneous urticaria-patients with auto-antibodies against IgE/IgE-receptor could induce TNFα and MMP-9 release from donor PBMCs, and if release levels could be used to assess severity and activity of chronic spontaneous urticaria (CSU). Met...

  16. Phage particles infecting branchial Rickettsiales-like organisms in banded carpet shell Polititapes virgineus (Bivalvia) from Galicia (NW Spain).

    Science.gov (United States)

    Darriba, S; Ruiz, M; López, C

    2012-09-12

    Basophilic intracellular prokaryotic-like colonies were observed in the gills of banded carpet shell Polititapes virgineus (= Tapes rhomboides) (Linnaeus, 1767) from a natural bed in Galicia (NW Spain). Light microscope observations suggested the presence of 2 types of colonies, but transmission electron microscopy revealed that these were the same Rickettsiales-like colonies, one infected and the other uninfected by phage particles. This is the first report of the presence of phage particles in Rickettsiales-like organisms in the gills of P. virgineus.

  17. Histologia da glândula de albúmen de Pomacea canaliculata (Lamarck, 1822 (Mollusca, Gastropoda, Pilidae

    Directory of Open Access Journals (Sweden)

    Eliane de Fátima Marques de Mesquita

    1990-01-01

    Full Text Available In this paper the authors give a histological analysis of the albumen gland of mature and immature females of Pomacea canaliculata (Lamarck, 1822. In the immaturity, tthe gland doesn't show any secretory activity, although in the maturity this activity is indicated by the presence of a homogeneous and eosinophilic material in its lumen. We could notice in the capsule gland of matures females a calcareous element is present as a granular and basophilic deposit. The albumen gland is envolved by a thin conjunctive layer that can be seen in van Gieson's coloured preparations.

  18. The skin: target organ in immunotoxicology of small-molecular-weight compounds.

    Science.gov (United States)

    Merk, H F; Sachs, B; Baron, J

    2001-01-01

    Immunotoxicology studies two different effects of xenobiotics: immunosuppression and dysregulation of immune responses leading to hypersensitivity or autoimmunity. The skin is a major target organ of immunotoxicity which is provoked by small-molecular-weight compounds. Methods may be helpful for immunotoxicological investigations and screenings for adverse effects of xenobiotics which are used for diagnosis or studies on the pathophysiology of skin disorders such as allergic contact dermatitis, cutaneous drug-allergic reactions or autoimmune diseases of the skin. Examples include well-designed patch tests, assays involving antigen-presenting cells such as dendritic cells, but also T lymphocytes, basophiles or keratinocytes.

  19. Xenotransplanted human prostate carcinoma (DU145) cells develop into carcinomas and cribriform carcinomas: ultrastructural aspects.

    Science.gov (United States)

    Gilloteaux, Jacques; Jamison, James M; Neal, Deborah R; Summers, Jack L; Taper, Henryk S

    2012-10-01

    Androgen-independent, human prostate carcinoma cells (DU145) develop into solid, carcinomatous xenotransplants on the diaphragm of nu/nu mice. Tumors encompass at least two poorly differentiated cell types: a rapidly dividing, eosinophilic cell comprises the main cell population and a few, but large basophilic cells able to invade the peritoneal stroma, the muscular tissue, lymph vessels. Poor cell contacts, intracytoplasmic lumina, and signet cells are noted. Lysosomal activities are reflected by entoses and programmed cell deaths forming cribriform carcinomas. In large tumors, degraded cells may align with others to facilitate formation of blood supply routes. Malignant cells would spread via ascites and through lymphatics.

  20. Systemic allergic reaction to pine nuts.

    Science.gov (United States)

    Nielsen, N H

    1990-02-01

    This case report describes a systemic reaction due to ingestion of pine nuts, confirmed by an open, oral provocation test. Skin prick testing with the aqueous allergen revealed an immediate positive prick test, and histamine release from basophil leukocytes to the aqueous allergen was demonstrated. Radioallergosorbent test demonstrated specific IgE antibodies to pine nuts. In a review of medical literature, we found no reports of either oral provocation tests confirming a systemic reaction due to ingestion of pine nuts or demonstration of specific IgE antibodies.

  1. Pin p 1 is a major allergen in pine nut and the first food allergen described in the plant group of gymnosperms.

    Science.gov (United States)

    Cabanillas, Beatriz; Crespo, Jesus F; Maleki, Soheila J; Rodriguez, Julia; Novak, Natalija

    2016-11-01

    This study aimed to report the complete sequence of a 2S albumin purified from pine nut and to analyze its allergenic properties. Individual recognition of this protein by serum IgE from pine nut-allergic patients was assessed. IgE cross-linking capacity was analyzed in a basophil activation test. Inhibition of IgE-binding and stability to heating was also assessed. The complete nucleotide sequence was obtained and a phylogenetic study was carried out. 2S albumin from pine nut (registered as Pin p 1.0101) was recognized by IgE of 75% of sera. The allergen was heat-stable and had a robust capacity to inhibit IgE-binding to whole pine nut extract. The IgE cross-linking capacity of Pin p 1 on basophils was also demonstrated. Despite the low homology of Pin p 1 sequence with other allergenic 2S albumins from angiosperms, Pin p 1 contains the typical skeleton of 8 cysteine residues, important for its α-helixes enriched structure.

  2. Food allergies: the basics.

    Science.gov (United States)

    Valenta, Rudolf; Hochwallner, Heidrun; Linhart, Birgit; Pahr, Sandra

    2015-05-01

    IgE-associated food allergy affects approximately 3% of the population and has severe effects on the daily life of patients-manifestations occur not only in the gastrointestinal tract but also affect other organ systems. Birth cohort studies have shown that allergic sensitization to food allergens develops early in childhood. Mechanisms of pathogenesis include cross-linking of mast cell- and basophil-bound IgE and immediate release of inflammatory mediators, as well as late-phase and chronic allergic inflammation, resulting from T-cell, basophil, and eosinophil activation. Researchers have begun to characterize the molecular features of food allergens and have developed chip-based assays for multiple allergens. These have provided information about cross-reactivity among different sources of food allergens, identified disease-causing food allergens, and helped us to estimate the severity and types of allergic reactions in patients. Importantly, learning about the structure of disease-causing food allergens has allowed researchers to engineer synthetic and recombinant vaccines.

  3. Glucocorticoid hypersensitivity as a rare but potentially fatal side effect of paediatric asthma treatment: a case report

    Directory of Open Access Journals (Sweden)

    Lehmann Sylvia

    2008-06-01

    Full Text Available Abstract Introduction Immediate-type hypersensitivity to glucocorticosteroids is rare but well known among allergists. Surprisingly, very few reports of glucocorticosteroid hypersensitivity in children exist although glucocorticosteroid treatment is particularly common in this age group. Case presentation We report the case of a 2-year-old boy who developed generalized urticaria, facial angio-oedema, nausea and severe dyspnoea after intravenous application of prednisolone-21-hydrogen succinate. Skin prick testing with prednisolone-21-hydrogen succinate elicited a positive result; no reactions were observed to prednisone, betamethasone or dexamethasone. While fluorescence enzyme immunoassay analysis revealed no specific IgE antibodies against prednisolone-21-hydrogen succinate, CD63-based basophil activation testing with the culprit drug prednisolone-21-hydrogen succinate was positive. In contrast, additional incubation of basophils with prednisone, betamethasone and dexamethasone did not elicit any significant response. Hence, we performed an oral provocation test with betamethasone and a titrated intravenous dexamethasone challenge. As both drugs were tolerated without any complications they were recommended for future treatment. Conclusion In a child with confirmed immediate-type hypersensitivity to glucocorticosteroids, it is still not possible to predict which glucocorticosteroid might be tolerated by solely relying on clinical history or results of skin and in vitro testing. Therefore, incremental glucocorticosteroid challenges under standardized clinical conditions remain necessary in order to facilitate a patient-tailored emergency treatment and to avoid severe reactions to glucocorticosteroids in these patients.

  4. Generation of an allergy vaccine by disruption of the three-dimensional structure of the cross-reactive calcium-binding allergen, Phl p 7.

    Science.gov (United States)

    Westritschnig, Kerstin; Focke, Margarete; Verdino, Petra; Goessler, Walter; Keller, Walter; Twardosz, Anna; Mari, Adriano; Horak, Friedrich; Wiedermann, Ursula; Hartl, Arnulf; Thalhamer, Josef; Sperr, Wolfgang R; Valent, Peter; Valenta, Rudolf

    2004-05-01

    The grass pollen allergen, Phl p 7, belongs to a family of highly cross-reactive calcium-binding pollen allergens. Because Phl p 7 contains most of the disease-eliciting epitopes of pollen-derived calcium-binding allergens, hypoallergenic variants were engineered according to the x-ray crystal structure of Phl p 7 for allergy vaccination. In three recombinant variants, amino acids essential for calcium binding were mutated, and two peptides comprising the N- and C-terminal half were obtained by synthetic peptide chemistry. As determined by circular dichroism analysis and size exclusion chromatography coupled to mass spectrometry, recombinant mutants showed altered structural fold and lacked calcium-binding capacity, whereas the two synthetic peptides had completely lost their structural fold. Allergic patients' IgE Ab binding was strongest reduced to the variant containing two mutations in each of the two calcium-binding sites and to the peptides. Basophil histamine release and skin test experiments in allergic patients identified the peptides as the vaccine candidates with lowest allergenic activity. Immunization of rabbits with the peptides induced IgG Abs that blocked allergic patients' IgE binding to Phl p 7 and inhibited allergen-induced basophil degranulation. Our results indicate that disruption of an allergen's three-dimensional structure represents a general strategy for the generation of hypoallergenic allergy vaccines, and demonstrate the importance of allergen-specific IgG Abs for the inhibition of immediate allergic symptoms.

  5. New insights in the structure and biology of the high affinity receptor for IgE (Fc epsilon RI) on human epidermal Langerhans cells.

    Science.gov (United States)

    Bieber, T; Kraft, S; Jürgens, M; Strobel, I; Haberstok, J; Tomov, H; Regele, D; de la Salle, H; Wollenberg, A; Hanau, D

    1996-10-01

    The recent structural and functional analysis of the high affinity receptor for IgE (Fc epsilon RI) expressed on human epidermal Langerhans cells (LC) revealed new aspects of the biology of this structure. In contrast to basophils and mast cells where this receptor seems to be expressed constitutively at a constant level, the expression of Fc epsilon RI on LC varies on the donor and the inflammatory environment of the cells and lacks the classical beta-chain. This also implies functional differences most probably related to the expression level. Although the signalling pathway seems to be similar to that of basophils or mast cells, LC from individuals with atopic dermatitis are fully activated by receptor ligation while LC from normal individuals fail to exhibit calcium mobilization under the same conditions. Finally, LC from normal and atopic individuals use Fc epsilon RI to maximize antigen uptake via specific IgE and subsequent presentation to T cells. Thus, Fc epsilon RI expressed on LC differs in terms of structure and function from that expressed on effector cells of anaphylaxis.

  6. Alpha-actinin is a new type of house dust mite allergen.

    Science.gov (United States)

    An, Su; Shen, Chuanbing; Liu, Xiaoyu; Chen, Lingling; Xu, Xuemei; Rong, Mingqiang; Liu, Zhigang; Lai, Ren

    2013-01-01

    Main indoor allergens for humans are from house dust mites. There are more than 30 allergens in Dermatophagoides farinae but only fourteen allergens have been identified from this mite including Der f 1-3, 6, 7, 10, 11, 13-18, and 22. A native allergen protein (Der f 24, 90 kDa) was purified from D. farinae by gel filtration and anionic exchange liquid chromatography combined with IgE immunodetection. Its primary structure was determined by Edman degradation, mass spectrometry analysis and cDNA cloning. Enzyme-linked immunosorbent assay inhibition tests (ELISA-IT), immunoblots, basophil activation test (BAT) and skin prick test (SPT) were performed to evaluate the allergenicity. It was identified as an alpha (α)-actinin containing a CaM-like domain with EF-hand motifs. Der f 24 reacted to sera from 85.4% (35/41) of patients on western blot analysis. It reduced ∼20% sera IgE reactivity to D. farinae extracts on a competitive ELISA. Eighty percent (8/10) of patients with D. farinae allergy showed positive reactions to Der f 24 in skin prick test. The expression of CD63 on basophils from patients was up-regulated by Der f 24 by ∼5.4-fold. Alpha-actinin was identified as a new type of house dust mite allergen. To the best of our knowledge, this is the first report of α-actinin as an allergen.

  7. Macroscopic and microscopic aspects of collared peccary and white-lipped peccary placenta.

    Science.gov (United States)

    Santos, T C; Dantzer, V; Jones, C J P; Oliveira, M F; Miglino, M A

    2006-01-01

    This study examines middle and late gestational placentae from 13 Tayassu tajacu (collared peccary) and 3 Tayassu pecari (white-lipped peccary), which are Artiodactyla belonging to the Family Tayassuidae. The chorionic sac of Tayassu species is diffuse and chorioallantoic. These epitheliochorial placentae show no trophoblast invasion into the uterine epithelium and there is interdigitation between fetal and maternal microvilli. Two distinct regions of the fetomaternal interface can be identified: the interareolar and the areolar regions. The uterine epithelium has eosinophilic cytoplasm with dispersed, basophilic and electron-dense granules. Trophoblast cells are irregularly cuboidal on top of the fetal ridges and columnar on troughs, where cells have cytoplasmic vesicles and large basal vacuoles, surrounded by whorls of smooth membranes. Capillaries indent the trophoblast cells forming a placental barrier 3 microm or less thick. The columnar uterine glandular epithelium has a subpopulation of granules staining with Perl's Prussian blue reaction, suggesting iron secretion. In areolar areas, the trophoblast cells show apical microvilli, a basophilic cytoplasm with electron-dense intracellular vacuoles and cisternae. The placenta can therefore be classified as non-deciduate. The ultrastructural aspects of this study reveal features that have not previously been described and extend our knowledge of functions relating to materno-fetal transport in these species.

  8. Evidence for novel age-dependent network structures as a putative primo vascular network in the dura mater of the rat brain

    Institute of Scientific and Technical Information of China (English)

    Ho-Sung Lee; Dai-In Kang; Seung Zhoo Yoon; Yeon Hee Ryu; Inhyung Lee; Hoon-Gi Kim; Byung-Cheon Lee; Ki Bog Lee

    2015-01-01

    With chromium-hematoxylin staining, we found evidence for the existence of novel age-depen-dent network structures in the dura mater of rat brains. Under stereomicroscopy, we noticed that chromium-hematoxylin-stained threadlike structures, which were barely observable in 1-week-old rats, were networked in specific areas of the brain, for example, the lateral lobes and the cerebella, in 4-week-old rats. In 7-week-old rats, those structures were found to have become larger and better networked. With phase contrast microscopy, we found that in 1-week-old rats, chromium-hematoxylin-stained granules were scattered in the same areas of the brain in which the network structures would later be observed in the 4- and 7-week-old rats. Such age-depen-dent network structures were examined by using optical and transmission electron microscopy, and the following results were obtained. The scattered granules fused into networks with increas-ing age. Cross-sections of the age-dependent network structures demonstrated heavily-stained basophilic substructures. Transmission electron microscopy revealed the basophilic substructures to be clusters with high electron densities consisting of nanosized particles. We report these data as evidence for the existence of age-dependent network structures in the dura mater, we discuss their putative functions of age-dependent network structures beyond the general concept of the dura mater as a supporting matrix.

  9. A correlative light microscopic, transmission and scanning electron microscopic study of the dorsum of human tongue.

    Science.gov (United States)

    Boshell, J L; Wilborn, W H; Singh, B B

    1980-01-01

    The dorsum of the human tongue has three types of papillae, filiform, fungiform and circumvallate. Some investigators have studied these by light and transmission electron microscopy. Since knowledge of the morphology through studies by scanning of the morphology through studies by scanning electron microscopy (SEM) is scant, this investigation was started with the purpose of studying human tongues at different ages. One fetal tongue and portions of three tongues from newborns were removed. Additional specimens were biopsied from the anterior region of three adult tongues. Samples were processed routinely for light microscopy, transmission electron microscoy (TEM) and scanning electron microscopy (SEM). Two distinct features were evident on the fetal tongue. The first was that the surface epithelial layer of the tongue appeared to be periderm. The second was that fungiform papillae began their development earlier than filiform. At birth, the putative periderm had disappeared and a few filiform papillae were observed. On the adult tongue, filiform papillae were numerous and were comprised of two cell populations. One cell population contained numerous keratohyalin granules (KHG). The KHG were two types, eosinophilic and basophilic. Ultrastructurally, the eosinophilic granules were less electron dense and larger in size than the basophilic KHG.

  10. Saiboku-to, a Kampo herbal medicine, inhibits LTC4 release from eosinophils

    Directory of Open Access Journals (Sweden)

    Koichi Hirai

    1998-01-01

    Full Text Available Saiboku-to (TJ-96, a traditional Kampo herbal formation, has been used in the treatment of bronchial asthma in Japan as an anti-allergy herbal medicine. We investigated the effect of TJ-96 on leukotriene (LTC4 release from eosinophils and basophils isolated from healthy volunteers. Pre-incubation of eosinophils with TJ-96 inhibited ionophore- or formyl-methionyl-leucyl-phenylalanine (FMLP-induced LTC4 generation by eosinophils in a dose-dependent fashion. The TJ-96 was more potent in the release by ionophore (IC50 = 60 mg/mL than the release induced by FMLP (IC50 = 300 mg/mL. Maximal inhibition was observed when eosinophils were pretreated with TJ-96 for 5 min. Although TJ-96 at high concentrations inhibited IgE-mediated histamine release from human basophils, inhibition of IgE-mediated LTC4 release was not statistically significant. The potent inhibitory activity was found in the extract of Glycyrrhiza root, one of the herbal components of TJ-96, but the inhibitory effects were not due to either glycyrrhizin or liquiritin, the main elements of the Glycyrrhiza root. These results raise the possibility that the clinical efficacy of TJ-96 is derived, at least in part, from its potent inhibitory effect on LTC4 release from eosinophils.

  11. Histamine release from gut mast cells from patients with inflammatory bowel diseases

    DEFF Research Database (Denmark)

    Nolte, Hendrik; Spjeldnæs, Nikolaj; Kruse, Aksel

    1990-01-01

    Inflammatory mediators from intestinal mast cells may serve as initiators of acute and delayed inflammation. Mast cell histamine release was measured in 19 patients with inflammatory bowel diseases using gut mast cells from enzymatically dispersed endoscopic forceps biopsy specimens of macroscopi......Inflammatory mediators from intestinal mast cells may serve as initiators of acute and delayed inflammation. Mast cell histamine release was measured in 19 patients with inflammatory bowel diseases using gut mast cells from enzymatically dispersed endoscopic forceps biopsy specimens...... colitis was increased compared with that in control subjects and patients with Crohn's disease, and the mast cell count obtained from inflamed tissue was greater than that of normal tissue. The study also shows the heterogeneity of the responsiveness of the histamine releasing cells to various...... secretagogues. Thus, mast cells released 0.4 (0.0-2.0) (median (range)) ng histamine per sample at anti-IgE challenge, and basophils were also anti-IgE responsive. In contrast, mast cells did not respond to FMLP but the corresponding basophils did. Gut mast cells released 0.3 (0.0-1.0) (median (range)) ng...

  12. Presentation and Diagnosis of Hypersensitivity to Platinum Drugs.

    Science.gov (United States)

    Caiado, Joana; Castells, Mariana

    2015-04-01

    Hypersensitivity reactions (HSRs) to platinum drugs are increasing due to their extensive use in a wide variety of malignancies and the repeated exposures in patients with increased life expectancy. Understanding the incidence of HSR to platinum drugs and associated risk factors can help with the diagnosis and may provide protection against severe HSRs. A thorough clinical history with identification of the typical and atypical symptoms, the relationship with the platin administration, and the number of previous exposures are the key to the diagnosis. An elevated serum tryptase at the time of the HSR indicates that IgE and/or mast cells/basophils were involved in the HSR. Skin testing to platinum drugs is a highly sensitive and specific diagnostic tool, which helps provide risk stratification and management recommendations. Platinum specific IgE measurement and basophil activation test (BAT) are emerging as new diagnostic tools and in combination with skin testing can help support the diagnosis and the cross-reactivity between the three most commonly used platinum drugs, namely carboplatin, cisplatin, and oxaliplatin.

  13. RARE BENIGN EYELID TUMOR IN CHILDREN (EPITHELIOMA OF MALHERBE, PILOMATRIXOMA, OR TRICHELEMMOMA

    Directory of Open Access Journals (Sweden)

    A. A. Ryabtseva

    2015-01-01

    Full Text Available Aim. To describe clinical manifestations of rare eyelid tumor (epithelioma Malherbe and to improve differential diagnosis of benign eyelid tumors in children. Patients and methods. We observed 8 children aged 3,5‑8 years (sex ratio was 1:1. In all cases, examination, palpation, surgical excision of the tumor with histological examination were performed. Results. Trichilemmoma, or pilomatricoma, was suggested from clinical manifestations. Epithelioma Malherbe was diagnosed by histology only. Microscopically, the tumor is surrounded by a capsule which includes two cell types. Peripheral basophilic cells are small cells with poor cytoplasm, indistinct borders, and deeply basophilic nucleus. Central shadow cells have a distinct border and a central unstained area. Islands of small basaloid epithelial cells with squamous cell focuses and cornification are embedded in the stroma. Epithelial lesions are often necrotized. Epithelial mass is surrounded by granulations with giant cells. Osseous trabeculae are often adjacent to necrotic lesions. Further follow-up revealed no complications or recurrences. Conclusions. Our observations and literature data suggest that epithelioma Malherbe is occured in 1.3 % of benign eyelid tumors in childern. Tumor growth is slow and non-invasive. 

  14. White spot syndrome virus isolates of tiger shrimp Penaeus monodon (Fabricious) in India are similar to exotic isolates as revealed by polymerase chain reaction and electron microscopy.

    Science.gov (United States)

    Mishra, S S; Shekhar, M S

    2005-07-01

    Microbiological analysis of samples collected from cases of white spot disease outbreaks in cultured shrimp in different farms located in three regions along East Coast of India viz. Chidambram (Tamil Nadu), Nellore (Andhra Pradesh) and Balasore (Orissa), revealed presence of Vibrio alginolyticus, Vibrio parahaemolyticus, and Aeromonas spp. but experimental infection trials in Penaeus monodon with these isolates did not induce any acute mortality or formation of white spots on carapace. Infection trials using filtered tissue extracts by oral and injection method induced mortality in healthy P. monodon with all samples and 100% mortality was noted by the end of 7 day post-inoculation. Histopathological analysis demonstrated degenerated cells characterized by hypertrophied nuclei in gills, hepatopancreas and lymphoid organ with presence of intranuclear basophilic or eosino-basophilic bodies in tubular cells and intercellular spaces. Analysis of samples using 3 different primer sets as used by other for detection of white spot syndrome virus (WSSV) generated 643, 1447 and 520bp amplified DNA products in all samples except in one instance. Variable size virions with mean size in the range of 110 x 320 +/- 20 nm were observed under electron microscope. It could be concluded that the viral isolates in India involved with white spot syndrome in cultured shrimp are similar to RV-PJ and SEMBV in Japan, WSBV in Taiwan and WSSV in Malaysia, Indonesia, Thailand, China and Japan.

  15. Occurrence, Development and Histochemical Characteristics of Lymphocystis in Cultured Japanese Flounder (Paralichthys olivaceus)

    Institute of Scientific and Technical Information of China (English)

    Sheng Xiuzhen(绳秀珍); Zhan Wenbin

    2004-01-01

    Occurrence and development of lymphocystis cells in flounder Paralichthys olivaceus, and some histochemical features are studied using light and electron microscopy and histochemical methods. Fibroblasts engulf virus particles forming phagocytosis vacuole in the cytoplasm. The infected cells proliferate and turn round and hypertrophied; basophilic inclusion bodies, Feulgen-positive, are present in the cytoplasm, and a hyaline capsule, containing acid ploysaccharide and protein, appears outside cell membrane. The mature lymphocystis cells possess numerous virus particles (200~220nm in diameter with a central core 120~140nm) and high electron-dense inclusions with budding virions at the surface in the cytoplasm, and show margination of chromatin. Moderate electron density particles (70~80nm in diameter) fill out perinuclear cisterna and arrange in crystalloid arrays in nucleus, liberated into the cytoplasm by rupture of nuclear membrane. No virus particles are observed in nucleus. Therefore, inclusion bodies are the sites of virus assembly and maturity, while replication processes of lymphocystis virus are involved in both nuclear and cytoplasmic phases. Besides, strongly basophilic link-like structures are present between adjacent cells or through connective tissue, and virus particles are observed in somewhere of the capsule, thus where may be the spread route of virus from one cell to another or to the environment. The senile lymphocystis cells release virus by the rupture of cells.

  16. A combination vaccine for allergy and rhinovirus infections based on rhinovirus-derived surface protein VP1 and a nonallergenic peptide of the major timothy grass pollen allergen Phl p 1.

    Science.gov (United States)

    Edlmayr, Johanna; Niespodziana, Katarzyna; Linhart, Birgit; Focke-Tejkl, Margarete; Westritschnig, Kerstin; Scheiblhofer, Sandra; Stoecklinger, Angelika; Kneidinger, Michael; Valent, Peter; Campana, Raffaela; Thalhamer, Josef; Popow-Kraupp, Theresia; Valenta, Rudolf

    2009-05-15

    Allergens and rhinovirus infections are among the most common elicitors of respiratory diseases. We report the construction of a recombinant combination vaccine for allergy and rhinovirus infections based on rhinovirus-derived VP1, the surface protein which is critically involved in infection of respiratory cells, and a nonallergenic peptide of the major grass pollen allergen Phl p 1. Recombinant hybrid molecules consisting of VP1 and a Phl p 1-derived peptide of 31 aa were expressed in Escherichia coli. The hybrid molecules did not react with IgE Abs from grass pollen allergic patients and lacked allergenic activity when exposed to basophils from allergic patients. Upon immunization of mice and rabbits, the hybrids did not sensitize against Phl p 1 but induced protective IgG Abs that cross-reacted with group 1 allergens from different grass species and blocked allergic patients' IgE reactivity to Phl p 1 as well as Phl p 1-induced basophil degranulation. Moreover, hybrid-induced IgG Abs inhibited rhinovirus infection of cultured human epithelial cells. The principle of fusing nonallergenic allergen-derived peptides onto viral carrier proteins may be used for the engineering of safe allergy vaccines which also protect against viral infections.

  17. Cutaneous mastocytomas in the neotenic caudate amphibians Ambystoma mexicanum (axolotl) and Ambystoma tigrinun (tiger salamander)

    Science.gov (United States)

    Harshbarger, J.C.; Chang, S.C.; DeLanney, L.E.; Rose, F.L.; Green, D.E.

    1999-01-01

    Spontaneous mastocytomas studied in 18 axolotls (Ambystoma mexicanum) and six tiger salamanders (Ambystoma tigrinum) were gray-white, uni- to multilobular cutaneous protrusions from 2mm to 2cm in diameter. Tumors were moderately cellular unencapsulated masses that usually infiltrated the dermis and hypodermis with the destruction of intervening tissues. Some tumors were invading superficial bundles of the underlying skeletal muscle. Tumors consisted of mitotically active cells derived from a single lineage but showing a range of differentiation. Immature cells had nearly smooth to lightly cleft or folded basophilic nuclei bordered by a band of cytoplasm with few cytoplasmic processes and containing a few small uniform eccentric granules. Mature cells had basophilic nuclei with deep clefts or folds and abundant eosinophilic cytoplasm with multiple long intertwining cytoplasmic extensions packed with metachromatic granules. The axolotls were old individuals from an inbred laboratory colony. The tiger salamanders were wild animals from a single polluted pond. They could have been old and inbred. Both groups were neotenic. These are the first mastocytomas discovered in cold-blooded animals.

  18. Cutaneous mastocytomas in the neotenic caudate amphibians Ambystoma mexicanum (axolotl) and Ambystoma tigrinum (tiger salamander).

    Science.gov (United States)

    Harshbarger, J C; Chang, S C; DeLanney, L E; Rose, F L; Green, D E

    1999-01-01

    Spontaneous mastocytomas studied in 18 axolotls (Ambystoma mexicanum) and six tiger salamanders (Ambystoma tigrinum) were gray-white, uni- to multilobular cutaneous protrusions from 2 mm to 2 cm in diameter. Tumors were moderately cellular unencapsulated masses that usually infiltrated the dermis and hypodermis with the destruction of intervening tissues. Some tumors were invading superficial bundles of the underlying skeletal muscle. Tumors consisted of mitotically active cells derived from a single lineage but showing a range of differentiation. Immature cells had nearly smooth to lightly cleft or folded basophilic nuclei bordered by a band of cytoplasm with few cytoplasmic processes and containing a few small uniform eccentric granules. Mature cells had basophilic nuclei with deep clefts or folds and abundant eosinophilic cytoplasm with multiple long intertwining cytoplasmic extensions packed with metachromatic granules. The axolotls were old individuals from an inbred laboratory colony. The tiger salamanders were wild animals from a single polluted pond. They could have been old and inbred. Both groups were neotenic. These are the first mastocytomas discovered in cold-blooded animals.

  19. Combination Chemotherapy With or Without PSC 833, Peripheral Stem Cell Transplantation, and/or Interleukin-2 in Treating Patients With Acute Myeloid Leukemia

    Science.gov (United States)

    2013-06-03

    Adult Acute Basophilic Leukemia; Adult Acute Eosinophilic Leukemia; Adult Acute Erythroid Leukemia (M6); Adult Acute Megakaryoblastic Leukemia (M7); Adult Acute Minimally Differentiated Myeloid Leukemia (M0); Adult Acute Monoblastic Leukemia (M5a); Adult Acute Monoblastic Leukemia and Acute Monocytic Leukemia (M5); Adult Acute Monocytic Leukemia (M5b); Adult Acute Myeloblastic Leukemia With Maturation (M2); Adult Acute Myeloblastic Leukemia Without Maturation (M1); Adult Acute Myeloid Leukemia With 11q23 (MLL) Abnormalities; Adult Acute Myeloid Leukemia With Del(5q); Adult Acute Myeloid Leukemia With Inv(16)(p13;q22); Adult Acute Myeloid Leukemia With t(16;16)(p13;q22); Adult Acute Myeloid Leukemia With t(8;21)(q22;q22); Adult Acute Myelomonocytic Leukemia (M4); Adult Erythroleukemia (M6a); Adult Pure Erythroid Leukemia (M6b); Childhood Acute Basophilic Leukemia; Childhood Acute Eosinophilic Leukemia; Childhood Acute Erythroleukemia (M6); Childhood Acute Megakaryocytic Leukemia (M7); Childhood Acute Minimally Differentiated Myeloid Leukemia (M0); Childhood Acute Monoblastic Leukemia (M5a); Childhood Acute Monoblastic Leukemia and Acute Monocytic Leukemia (M5); Childhood Acute Monocytic Leukemia (M5b); Childhood Acute Myeloblastic Leukemia With Maturation (M2); Childhood Acute Myeloblastic Leukemia Without Maturation (M1); Childhood Acute Myelomonocytic Leukemia (M4); Childhood Myelodysplastic Syndromes; de Novo Myelodysplastic Syndromes; Untreated Adult Acute Myeloid Leukemia; Untreated Childhood Acute Myeloid Leukemia and Other Myeloid Malignancies

  20. Syrian hamsters (Mesocricetus auratus) with simultaneous intestinal Giardia sp., Spironucleus sp., and trichomonad infections.

    Science.gov (United States)

    Sheppard, Barbara J; Stockdale Walden, Heather D; Kondo, Hirotaka

    2013-11-01

    A commercial facility producing hamsters with a history of infection by dwarf tapeworm (Hymenolepis nana) submitted 15 animals for necropsy and postmortem parasitological and microscopic examination. No tapeworms were detected grossly or microscopically. Fecal examination including gastrointestinal mucosal smears demonstrated mixed intestinal bacteria and low numbers of Giardia sp. Histologic examination of small intestine demonstrated filling of the small intestinal crypts by large numbers of 7-9 µm × 3 µm, rod to crescent or teardrop-shaped flagellates consistent with Spironucleus sp. These organisms had two 1-µm, basophilic, oval nuclei and multiple superficial flagella-like structures. Much larger 10-15 µm × 8-10 µm, oval to pear-shaped organisms were also present in lower numbers and usually located with the crypts. These larger flagellates had multiple flagella and a basophilic rod-shaped nucleus. The larger flagellates included Giardia sp., which had an intimate interface with the surface of the mucosal epithelium, bilaterally symmetry, and binucleation. Lower numbers of trichomonads were also present and were distinguished by an undulating surface membrane and a single nucleus. The mucosa was hyperplastic and moderately inflamed. Although the tapeworm infection was resolved, diagnosis of multiple intestinal flagellates by fecal examination is complicated by the varying sensitivity and diagnostic accuracy of different types of fecal analysis for different flagellate types. Key differences in the morphology and location of the different types of flagellates as observed by histology of intestinal tissues provide important additional diagnostic information to distinguish trichomonads, Spironucleus sp., and Giardia sp.

  1. Novel strategy to create hypoallergenic peanut protein-polyphenol edible matrices for oral immunotherapy.

    Science.gov (United States)

    Plundrich, Nathalie J; Kulis, Mike; White, Brittany L; Grace, Mary H; Guo, Rishu; Burks, A Wesley; Davis, Jack P; Lila, Mary Ann

    2014-07-23

    Peanut allergy is an IgE-mediated hypersensitivity. Upon peanut consumption by an allergic individual, epitopes on peanut proteins bind and cross-link peanut-specific IgE on mast cell and basophil surfaces triggering the cells to release inflammatory mediators responsible for allergic reactions. Polyphenolic phytochemicals have high affinity to bind proteins and form soluble and insoluble complexes with unique functionality. This study investigated the allergenicity of polyphenol-fortified peanut matrices prepared by complexing various polyphenol-rich plant juices and extracts with peanut flour. Polyphenol-fortified peanut matrices reduced IgE binding to one or more peanut allergens (Ara h 1, Ara h 2, Ara h 3, and Ara h 6). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) suggested changes in secondary protein structure. Peanut protein-cranberry polyphenol fortified matrices triggered significantly less basophil degranulation than unmodified flour in an ex vivo assay using human blood and less mast cell degranulation when used to orally challenge peanut-allergic mice. Polyphenol fortification of peanut flour resulted in a hypoallergenic matrix with reduced IgE binding and degranulation capacity, likely due to changes in protein secondary structure or masking of epitopes, suggesting potential applications for oral immunotherapy.

  2. Alpha-actinin is a new type of house dust mite allergen.

    Directory of Open Access Journals (Sweden)

    Su An

    Full Text Available Main indoor allergens for humans are from house dust mites. There are more than 30 allergens in Dermatophagoides farinae but only fourteen allergens have been identified from this mite including Der f 1-3, 6, 7, 10, 11, 13-18, and 22. A native allergen protein (Der f 24, 90 kDa was purified from D. farinae by gel filtration and anionic exchange liquid chromatography combined with IgE immunodetection. Its primary structure was determined by Edman degradation, mass spectrometry analysis and cDNA cloning. Enzyme-linked immunosorbent assay inhibition tests (ELISA-IT, immunoblots, basophil activation test (BAT and skin prick test (SPT were performed to evaluate the allergenicity. It was identified as an alpha (α-actinin containing a CaM-like domain with EF-hand motifs. Der f 24 reacted to sera from 85.4% (35/41 of patients on western blot analysis. It reduced ∼20% sera IgE reactivity to D. farinae extracts on a competitive ELISA. Eighty percent (8/10 of patients with D. farinae allergy showed positive reactions to Der f 24 in skin prick test. The expression of CD63 on basophils from patients was up-regulated by Der f 24 by ∼5.4-fold. Alpha-actinin was identified as a new type of house dust mite allergen. To the best of our knowledge, this is the first report of α-actinin as an allergen.

  3. In Vitro Research Tools in the Field of Human Immediate Drug Hypersensitivity and Their Present Use in Small Animal Veterinary Medicine

    Directory of Open Access Journals (Sweden)

    Lavergne S. Lavergne

    2016-12-01

    Full Text Available Drug hypersensitivity reactions (DHR are immune-mediated idiosyncratic adverse drug events. Type I DHR are often referred to as “immediate” and involve B lymphocyte-secreted IgE that bind to the membrane of basophils and mast cells, inducing their degranulation. This review presents various in vitro tests that were developed in the field of human type I HS and implemented as clinical diagnostic tools in human cases of immediate DHR. The respective strengths and weaknesses of each test will be discussed in parallel of validation data such as specificity and sensitivity whenever available. Some of them have also been used as diagnostic tools in veterinary medicine, but not in cases of immediate DHR. Most of these diagnostic tools can be categorized into humoral and cellular tests. The former tests measure serum concentrations of factors, such as histamine, tryptase, and drug-specific IgE. The latter assays quantify markers of drug-induced basophil activation or drug-specific lymphocyte proliferation. Pharmacogenetic markers have also been investigated in immediate DHR, but not as extensively as in non-immediate ones. Throughout, practical aspects and limitations of the tests, as well as sensitivity and specificity parameters, will be presented. In addition, the experience of veterinary medicine with these diagnostic tools will be summarized. However, to date, none of them has ever been reported in a veterinary case of type I DHR.

  4. A fluorescence-based centrifugal microfluidic system for parallel detection of multiple allergens

    Science.gov (United States)

    Chen, Q. L.; Ho, H. P.; Cheung, K. L.; Kong, S. K.; Suen, Y. K.; Kwan, Y. W.; Li, W. J.; Wong, C. K.

    2010-02-01

    This paper reports a robust polymer based centrifugal microfluidic analysis system that can provide parallel detection of multiple allergens in vitro. Many commercial food products (milk, bean, pollen, etc.) may introduce allergy to people. A low-cost device for rapid detection of allergens is highly desirable. With this as the objective, we have studied the feasibility of using a rotating disk device incorporating centrifugal microfluidics for performing actuationfree and multi-analyte detection of different allergen species with minimum sample usage and fast response time. Degranulation in basophils or mast cells is an indicator to demonstrate allergic reaction. In this connection, we used acridine orange (AO) to demonstrate degranulation in KU812 human basophils. It was found that the AO was released from granules when cells were stimulated by ionomycin, thus signifying the release of histamine which accounts for allergy symptoms [1-2]. Within this rotating optical platform, major microfluidic components including sample reservoirs, reaction chambers, microchannel and flow-control compartments are integrated into a single bio-compatible polydimethylsiloxane (PDMS) substrate. The flow sequence and reaction time can be controlled precisely. Sequentially through varying the spinning speed, the disk may perform a variety of steps on sample loading, reaction and detection. Our work demonstrates the feasibility of using centrifugation as a possible immunoassay system in the future.

  5. Bone marrow contribution to eosinophilic inflammation

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    Denburg Judah A

    1997-01-01

    Full Text Available Allergen-induced bone marrow responses are observable in human allergic asthmatics, involving specific increases in eosinophil-basophil progenitors (Eo/B-CFU, measured either by hemopoietic assays or by flow cytometric analyses of CD34-positive, IL-3Ralpha-positive, and/or IL-5-responsive cell populations. The results are consistent with the upregulation of an IL-5-sensitive population of progenitors in allergen-induced late phase asthmatic responses. Studies in vitro on the phenotype of developing eosinophils and basophils suggest that the early acquisition of IL-5Ralpha, as well as the capacity to produce cytokines such as GM-CSF and IL-5, are features of the differentiation process. These observations are consistent with findings in animal models, indicating that allergen-induced increases in bone marrow progenitor formation depend on hemopoietic factor(s released post-allergen. The possibility that there is constitutive marrow upregulation of eosinophilopoiesis in allergic airways disease is also an area for future investigation.

  6. Optineurin and amyotrophic lateral sclerosis.

    Science.gov (United States)

    Maruyama, Hirofumi; Kawakami, Hideshi

    2013-07-01

    Amyotrophic lateral sclerosis is a devastating disease, and thus it is important to identify the causative gene and resolve the mechanism of the disease. We identified optineurin as a causative gene for amyotrophic lateral sclerosis. We found three types of mutations: a homozygous deletion of exon 5, a homozygous Q398X nonsense mutation and a heterozygous E478G missense mutation within its ubiquitin-binding domain. Optineurin negatively regulates the tumor necrosis factor-α-induced activation of nuclear factor kappa B. Nonsense and missense mutations abolished this function. Mutations related to amyotrophic lateral sclerosis also negated the inhibition of interferon regulatory factor-3. The missense mutation showed a cyotoplasmic distribution different from that of the wild type. There are no specific clinical symptoms related to optineurin. However, severe brain atrophy was detected in patients with homozygous deletion. Neuropathologically, an E478G patient showed transactive response DNA-binding protein of 43 kDa-positive neuronal intracytoplasmic inclusions in the spinal and medullary motor neurons. Furthermore, Golgi fragmentation was identified in 73% of this patient's anterior horn cells. In addition, optineurin is colocalized with fused in sarcoma in the basophilic inclusions of amyotrophic lateral sclerosis with fused in sarcoma mutations, and in basophilic inclusion body disease. These findings strongly suggest that optineurin is involved in the pathogenesis of amyotrophic lateral sclerosis.

  7. A COMPARATIVE STUDY OF THE OVINE HAEMOGRAM: CELL-DYN 3500 VERSUS MANUAL METHODS

    Directory of Open Access Journals (Sweden)

    Labrini V. Athanasiou

    2013-01-01

    Full Text Available Manual hematology techniques traditionally applied in farm animal medicine are time consuming and labor-intensive, especially when large numbers of samples have to be processed. As a result, several automated hematology instruments have been developed for use in these species. An automated hematology analyzer (Abbott Cell-Dyne 3500 system was used in performing a complete blood cell count and differential counts of white blood cells in sheep blood samples. The system was compared with basic manual hematologic techniques. A linear regression was used to assess correlation between the two methods. Correlation coefficients (R2 were good for the hematocrit, the total white blood cell count, the neutrophils, the lymphocytes and the platelet count, while a poor correlation existed in monocytes, eosinophils and basophils. The automated and the manual technique were also compared in terms of sensitivity using the Sensitivity Ratio (SR. The automated analyzer was slightly more sensitive than the manual technique for all parameters tested except for monocytes, eosinophils and basophils were the difference was greater and the automated analyzer was 1.5, 2.5 and 2 times more reliable, respectively. The method bias was also calculated. It seems that the overall performance of the automated analyzer justifies its utilization in sheep blood analysis, although as for any analyzer used in any species, a stained blood film evaluation remains an indispensable technique to confirm the results being reported by the automated analyzer and provide additional information for the ovine haemogram.

  8. Apprecation on the HRBT was Induced by Anti- IgE with Efficiency of Cross- Link%交联效应在抗IgE抗体诱导致敏嗜碱粒细胞脱颗粒实验中的应用

    Institute of Scientific and Technical Information of China (English)

    朱玲; 陆惠民; 余传星; 何小丽

    2002-01-01

    目的将二个IgE分子在靶细胞表面通过双价配体产生交联,这一诱导Ⅰ型变态反应的理论,运用到由抗IgE诱导致敏嗜碱粒细胞组胺释放试验(histamine release of basophils test,HRBT)中,以获得组胺(histamine,HA)的高释放率.方法将IgE的浓度与抗IgE的浓度按不同的比例进行组合,用微量HA荧光测定法观察各配对组对人血细胞中嗜碱粒细胞(basophils,BOS)释放HA的影响.结果交联组的HA释放率明显高于对照组,t=4.604,P<0.01.结论当IgE:抗IgE的浓度=2:1时,BOS的HA释放率随该比值的增高而增高;若该比值倒置,则HA释放受到抑制.

  9. Chronic autoimmune urticaria : Where we stand ?

    Directory of Open Access Journals (Sweden)

    Goh C

    2009-01-01

    Full Text Available It is well-recognized that 30-40% of chronic idiopathic urticaria is autoimmune in nature. Chronic autoimmune urticaria is caused by anti-FcåRI and less frequently, by anti-IgE autoantibodies that lead to mast cell and basophil activation, thereby giving rise to the release of histamine and other proinflammatory mediators. Activation of the classical complement pathway and formation of C5a are important in dermal mast cell activation. C5a is also a neutrophil and eosinophil chemoattractant. Chronic autoimmune urticaria has been found to be associated with autoimmune thyroid disease. The autologous serum skin test is used as a screening test for chronic autoimmune urticaria and has a sensitivity and specificity of about 70 and 80%, respectively. The current gold standard diagnostic test is the basophil histamine release assay. The treatment of chronic autoimmune urticaria, as in chronic idiopathic urticaria, is with H1 antihistamines. Oral corticosteroids may be used during acute flares. Refractory cases have been shown to respond to cyclosporine and other immunomodulators. The prevalence of chronic autoimmune urticaria in Singapore is similar to that reported in Western countries at about 42%. The presence of thyroid autoimmunity appears to be higher than reported, with 22.5% of patients with chronic idiopathic urticaria here, exhibiting presence of thyroid autoantibodies.

  10. Limited influence of aspirin intake on mast cell activation in patients with food-dependent exercise-induced anaphylaxis: comparison using skin prick and histamine release tests.

    Science.gov (United States)

    Fukunaga, Atsushi; Shimizu, Hideki; Tanaka, Mami; Kikuzawa, Ayuko; Tsujimoto, Mariko; Sekimukai, Akiko; Yamashita, Junji; Horikawa, Tatsuya; Nishigori, Chikako

    2012-09-01

    Food-dependent exercise-induced anaphylaxis (FDEIA) is a severe systemic syndrome induced by physical exercise after ingesting causative food. Aspirin is a well-known trigger for anaphylaxis in patients with FDEIA. Possible mechanisms by which symptoms are aggravated by aspirin include enhanced antigen absorption and mast cell activation. The aim of this study was to determine whether aspirin intake has an influence on mast cell/basophil activation in patients with FDEIA. Provocation tests revealed that adding aspirin to the causative food challenge in 7 of 9 (77.8%) patients with FDEIA provoked symptoms. In most cases, pretreatment with aspirin did not enhance skin tests (71.4%) or histamine release tests (88.9%) with food allergen challenges. The study confirms that histamine release and skin prick tests can be adjunctive tools for diagnosing FDEIA. In addition, our results suggest that exacerbation of FDEIA symptoms by aspirin is not mediated by direct effects of aspirin on mast cell/basophil activation.

  11. Enzymatic hydrolysis: a method in alleviating legume allergenicity.

    Science.gov (United States)

    Kasera, Ramkrashan; Singh, A B; Lavasa, S; Prasad, Komarla Nagendra; Arora, Naveen

    2015-02-01

    Legumes are involved in IgE mediated food allergy in many countries. Avoidance of allergenic food is the only way to avoid symptomatic reaction. The present study investigated the effect of enzymatic hydrolysis on the allergenicity of three legumes - kidney bean (Phaseolus vulgaris), black gram (Vigna mungo) and peanut (Arachis hypogaea). Soluble protein extracts of the study legumes were sequentially treated by Alcalase(®) and Flavourzyme(®). Allergenicity of hydrolysates was then determined by ELISA, immunoblot, stripped basophil histamine release and skin prick test (SPT). Hydrolysis resulted in the loss of all IgE binding fractions determined by immunoblot in the three legumes. Specific IgE binding in ELISA was reduced by 62.2 ± 7.7%, 87.1 ± 9.6% and 91.8 ± 7.2% in the hydrolysates of kidney bean, black gram and peanut, respectively (p release of histamine was decreased significantly when sensitized basophils were challenged with hydrolysates as compared to raw extracts. Significant reduction in the biopotency of hydrolysates was also observed in SPT where only 1/10 kidney bean-sensitive individuals, 2/6 black gram-sensitive individuals and 1/7 peanut-sensitive individuals were found positive to their respective hydrolysates. In conclusion, enzymatic hydrolysis is effective in attenuating allergenicity of legume proteins and may be employed for preparing hypoallergenic food extracts.

  12. [Automated measurement of reticulocyte count by flow cytometry. II: Analysis of the blood containing abnormal erythrocytes or giant platelets].

    Science.gov (United States)

    Oyamatsu, T; Shimizu, N; Takeuchi, K; Yamamoto, M; Kawai, Y; Watanabe, K; Iri, H

    1989-07-01

    We have examined the influence of erythrocytes containing inclusion bodies, nucleated red cells or giant platelets on the measurement of reticulocyte count by automated machine, R-1000. Correlation of the reticulocyte count between automated and conventional method was extremely good in the blood containing red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling . However, correlation was poor when the sample contained the nucleated red cells. Reticulocyte count was decreased in the blood with significant amounts of nucleated red cells. Since nucleated red cells themselves are not counted as reticulocytes in the machine, this was considered to be due to increased young reticulocytes which frequently appeared with nucleated red cells. Both cold agglutinated red cells and giant platelets apparently influenced the reticulocyte count by the R-1000. These results suggest that red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling do not influence the automatic counting of reticulocytes. Although nucleated red cells, cold agglutinated red cells and giant platelets affected the reticulocyte count, the machine shows abnormal flags in most of above cases (except highly agglutinated red cells), so that one can recount reticulocytes by conventional method. We conclude the machine can safely count the reticulocytes even in the blood containing abnormal red cells or platelets.

  13. Repeat-dose toxicology evaluation in cynomolgus monkeys of AVI-4658, a phosphorodiamidate morpholino oligomer (PMO) drug for the treatment of duchenne muscular dystrophy.

    Science.gov (United States)

    Sazani, Peter; Ness, Kirk P Van; Weller, Doreen L; Poage, Duane W; Palyada, Kiran; Shrewsbury, Stephen B

    2011-05-01

    AVI-4658 is a phosphorodiamidate morpholino oligomer (PMO) drug designed to restore dystrophin expression in a subset of patients with Duchenne muscular dystrophy (DMD). Previous reports demonstrated this clinical proof-of-principle in patients with DMD following intramuscular injection of AVI-4658. This preclinical study evaluated the toxicity and toxicokinetic profile of AVI-4658 when administered either intravenously (IV) or subcutaneously (SC) to cynomolgus monkeys once weekly over 12 weeks, at doses up to the maximum feasible dose of 320 mg/kg per injection. No drug-related effects were noted on survival, clinical observations, body weight, food consumption, opthalmoscopic or electrocardiographic evaluations, hematology, clinical chemistry, urinalysis, organ weights, and macroscopic evaluations. Drug-related microscopic renal effects were dose-dependent, apparently reversible, and included basophilic granules (minimal), basophilic tubules (minimal to moderate), and tubular vacuolation (minimal to mild). These data establish the tolerability of AVI-4658 at doses up to and including the maximum feasible dose of 320 mg/kg by IV bolus or SC injection.

  14. Changes in white blood pictures and some biochemical parameters of dairy cows in peripartum period and early lactation

    Directory of Open Access Journals (Sweden)

    Mirjana Joksimović Todorović

    2012-06-01

    Full Text Available The objective of this research was to determine the changes in the number of cells of white blood line and some biochemical parameters: concentration of glucose, concentration of non-esterified fatty acids (NEFA, activity of enzymes of aspartat aminotransferase (AST, alanin aminotransferase (ALT and gamma glutamyltranferase (GGT, in 17 Holstein-Friesian breed dairy cows in peripartum period and early lactation (on d 15 prepartum, on the day of parturition and on d 15, 30, and 45 postpartum. The highest values of total leukocytes count, neutrophiles, monocytes and basophiles count were recorded on the day of parturition, and then decreased, so that the minimum value established on d 45 of lactation. The lymphocytes count was the lowest on the day of parturation, while the eosinophils and basophiles counts were significantly different in observed time intervals. The concentration of glucose ranged from 3.07 mmol/L on d 15 prepartum to 2.71 mmol/L on d 15 postpartum. A high concentration of non-esterified fatty acids (NEFA in the postpartum period is a consequence of changes in energy balance and more intensive process of lypolisis in fat tissue, due to energy disbalance at the beginning of lactation. Established activity of enzymes AST, ALT and GGT was approximately the same in studied time intervals and was within physiological limits. The results of hematological and biochemical analysis in this paper do not indicate the development of postpartum ailments, because the investigated parameters were within physiological limits.

  15. Leukocyte Populations are Associated with Heart Rate Variability After a Triathlon.

    Science.gov (United States)

    Cruz, Germán Hernández; Orellana, José Naranjo; Taraco, Adrián Rosas; Colmenero, Blanca Rangel

    2016-12-01

    The purpose of this study was to analyze cellular immune components and their association with heart rate variability in triathlon athletes. Twelve athletes were included (age 36.41 ± 5.57 years, body mass 81.84 ± 10.97 kg) and blood samples were taken one week before, immediately, at 2 and 48 hours, and one week after competition. Total lymphocytes and their subpopulations, neutrophils, basophils, eosinophils and monocytes were analyzed. At the same time, heart rate variability was recorded for 30 minutes using Polar Team2®. A significant difference between lymphocyte subpopulations and heart rate variability was found in the different study periods. A positive correlation was found between total lymphocytes and rMSSD (r = .736, p <0.05), CD3+ and rMSSD (r = .785, p <0.05), and CD4+ and rMSSD (r = .795, p < 0.05) at the end of the competition. After one week of competition, a negative correlation was found between eosinophils and MRR, SDNN, pNN50, and rMSSD (p <0.01); and basophils and MRR, SDNN, pNN50, and rMSSD (p <0.01); while a positive correlation was found between CD19+ (B cells) and pNN50 (r = .678, p <0.05). Our results suggest that it is possible to predict the effect of training with regard to the athlete's performance.

  16. Leukocyte Populations are Associated with Heart Rate Variability After a Triathlon

    Directory of Open Access Journals (Sweden)

    Cruz Germán Hernández

    2016-12-01

    Full Text Available The purpose of this study was to analyze cellular immune components and their association with heart rate variability in triathlon athletes. Twelve athletes were included (age 36.41 ± 5.57 years, body mass 81.84 ± 10.97 kg and blood samples were taken one week before, immediately, at 2 and 48 hours, and one week after competition. Total lymphocytes and their subpopulations, neutrophils, basophils, eosinophils and monocytes were analyzed. At the same time, heart rate variability was recorded for 30 minutes using Polar Team2®. A significant difference between lymphocyte subpopulations and heart rate variability was found in the different study periods. A positive correlation was found between total lymphocytes and rMSSD (r = .736, p <0.05, CD3+ and rMSSD (r = .785, p <0.05, and CD4+ and rMSSD (r = .795, p < 0.05 at the end of the competition. After one week of competition, a negative correlation was found between eosinophils and MRR, SDNN, pNN50, and rMSSD (p <0.01; and basophils and MRR, SDNN, pNN50, and rMSSD (p <0.01; while a positive correlation was found between CD19+ (B cells and pNN50 (r = .678, p <0.05. Our results suggest that it is possible to predict the effect of training with regard to the athlete's performance.

  17. Role of interleukin-18 in the pathophysiology of allergic diseases.

    Science.gov (United States)

    Sanders, Nathan L; Mishra, Anil

    2016-12-01

    Interleukin (IL)-18 is an IL-1 family cytokine expressed by macrophages, dendritic cells, epithelial cells, and keratinocytes and is implicated in various aspects of both the innate and adaptive immune systems. IL-18 signals similar to IL-1β intracellularly to activate gene transcription. Since its discovery, IL-18 has been demonstrated to play a key role in pathogen defense from helminths and some bacteria. Recently however, evidence has accumulated that IL-18 expression is increased in many presentations of allergic disease. A pathologic role for IL-18 includes stimulating mast cell and basophil degranulation, recruiting granulocytes to sites of inflammation, increasing cytotoxic activity of natural killer (NK) and NK-T cells, inducing Immunoglobulin (Ig)E production and isotype switching, and affecting a broad range of T cells to promote a type II helper T cell (Th2) response. Evidence and importance of these effects are presented, including novel results from our lab implicating IL-18 in the direct expansion of mast cells, basophils, and other myeloid-lineage cells from bone-marrow precursors. The development of urticaria, asthma, dermatitis, rhinitis, and eosinophilic disorders all have demonstrated correlations to increased IL-18 levels either in the tissue or systemically. IL-18 represents a novel site of immune regulation in not only allergic conditions, but also autoimmune diseases and other instances of aberrant immune functioning. Diagrammatic summarized abstract for readers convinance is presented in Fig. 1.

  18. Diagnostic Significance of BAT in Anaphylaxis to Non-ionic Contrast Media%BAT在非离子型造影剂过敏反应中的诊断价值

    Institute of Scientific and Technical Information of China (English)

    张皓月; 许素军; 唐笑先; 牛记军; 郭相杰; 高彩荣

    2015-01-01

    Objective To investigate the diagnostic significance of basophil activation test (B A T) in ana-phylaxis to non-ionic contrast media through testing the content of CD 63, m ast cell-carboxypeptidase A 3 (M C-CPA 3), and term inal com plem ent com plex SC5b-9 of the individuals by testing their levels in the norm al im m une group and the anaphylaxis groups to β-lactam drugs and non-ionic contrast media. Methods The CD 63 expression of basophilic granulocyte in blood w as detected by flow cytom etry. The levels of M C-CPA 3 in blood serum and SC5b-9 in blood plasm a w ere detected by ELISA . Results The CD 63 expression of basophilic granulocyte in blood, the levels of M C-CPA 3 and SC5b-9 of anaphylaxis to non-ionic contrast media and β-lactam drugs w ere significantly higher than that in norm al im m une group (P<0.05). Conclusion There is activation of basophilic granulocytes, m ast cells and com plem ent system in anaphylaxis to non-ionic contrast media. B A Tcan be used to diagnose the anaphylaxis to non-ionic contrast media.%目的:通过测定免疫正常人、β-内酰胺类药物过敏者以及非离子型造影剂过敏者血液中CD63、肥大细胞羧肽酶A3(MC-CPA3)、人末端补体复合物SC5b-9,探讨嗜碱性粒细胞活化试验(basophil activation test,BAT)在非离子型造影剂过敏反应中的诊断价值。方法采用流式细胞术测定全血中嗜碱性粒细胞CD63表达率。采用ELISA法测定血清中MC-CPA3、血浆中SC5b-9含量。结果非离子型造影剂和β-内酰胺类药物过敏者血液中嗜碱性粒细胞CD63表达率、MC-CPA3以及SC5b-9含量均较免疫正常人升高(P<0.05)。结论非离子型造影剂过敏者血液中嗜碱性粒细胞和肥大细胞发生活化,补体系统的激活也参与了非离子型造影剂过敏反应的发生。流式细胞术分析BAT可作为非离子型造影剂过敏反应的诊断方法。

  19. In vitro and in vivo anti-allergic effects of Arctium lappa L.

    Science.gov (United States)

    Knipping, Karen; van Esch, Elisabeth C A M; Wijering, Selva C; van der Heide, Sicco; Dubois, Anthony E; Garssen, Johan

    2008-11-01

    The discovery of drugs that can be used for the treatment of allergic disease is important in human health. Arctium lappa Linne (Compositae) (AL) has been used as a traditional medicine in Brazil and throughout Asia and is known to have an anti-inflammatory effect. In this study, the inhibitory effects of AL on degranulation and the release of mediators as well as on inhibition of cys-leukotriene biosynthesis by basophils were investigated. AL was selected out of 10,000 herbal extracts in a set-up for high throughput screening in which the degree of degranulation was monitored by the release of beta-hexosaminidase from rat basophil leukemia (RBL-2H3) cells. The AL extract significantly reduced degranulation and biosynthesis of cys-leukotrienes of human basophils in peripheral blood mono-nuclear cells (PBMCs) (50% inhibitory concentration [IC(50)] = 8.3 and 11.4 microg/ml, respectively). Viability and metabolic activity of the PBMCs were not affected. Although arctiin, the active component of AL that has been described in the literature, was not able to reduce degranulation in RBL-2H3 cells, a single high-performance liquid chromatography (HPLC) fraction from the AL extract inhibited beta-hexosaminidase release (IC(50) = 22.2 microg/ml). Topical administration of an aqueous extract of AL (5 mg/ear) on the ear of whey-sensitized mice 4 hrs before challenge with whey in the ear inhibited acute ear swelling by 50% in an in vivo cow's milk allergic model. The extract had no effect in this model when administered orally. In conclusion, the active component present in the active HPLC fraction of the AL extract was able to significantly reduce the release of inflammatory mediators through inhibition of degranulation and cys-leukotriene release in vitro. In addition, this active component was able to inhibit acute skin response in mice in vivo, indicating that AL is a very promising natural component for use in anti-allergic treatment.

  20. 杂交鲍苗种“病毒性萎缩症(AbSV)”病理学研究%Studies on Pathology of “Shriveling Syndrome Associated Virus(AbSV)” in Juvenile Hybrid Abalone

    Institute of Scientific and Technical Information of China (English)

    杨小强

    2013-01-01

    从福州地区患病杂交鲍苗种组织中分离到一种球形病毒用以感染健康鲍,通过对感染发病的鲍苗进行光镜和电镜观察,发现:病毒主要在消化腺嗜碱性细胞的细胞核中发生;消化腺消化细胞和嗜碱性细胞细胞核异形、染色质固缩,内质网扩张,线粒体嵴断裂、消失;消化腺小叶基膜肌纤维断裂、消失;消化道肠道具微绒毛的柱状细胞和纤毛柱状细胞发生自溶现象;足部肌细胞核浓缩,染色质固缩、边集,肌纤维断裂出现空泡.据此推测,病毒以鲍消化腺嗜碱性细胞为靶细胞,造成消化腺和胃、肠道等消化系统病变,机体因长期营养不良而萎缩,最后因全身性感染出现死亡.%Pathological analysis was done on juvenile hybrid abalone (Haliotisdiscus discus Haliotis discus hannai)artificially infected by a spherical virus isolated from the diseased ones in Fuzhou.Examination of artificially infected juvenile hybrid abalone by optical and electron microscopy showed that the virion mainly developed in nucleus of basophilic cells in digestive gland,and led to nuclear heteromorphosis,chromatin condensation,endoplasmic reticulum expansion,and mitochondria cristae breakage in digestive gland cell and basophilic cell.Meanwhile,broken digestive gland lobule basal lamina muscle fibers,autolysis and pyknosis of microvillus and cilium columnar cell in digestive gland intestinal tract were observed.In foot muscle cells,muscle fibers were broken and vesicles emerged,together with chromatin condensation and margination.These results suggested that the digestive gland basophilic cells maybe the special target cell of this virus,the infection could cause lesion in digestive gland and digestive organs such as stomach,intestines.Atrophy due to long-term malnutrition and systemic infection would finally led to death.

  1. Mast cell proteases as pharmacological targets.

    Science.gov (United States)

    Caughey, George H

    2016-05-05

    Mast cells are rich in proteases, which are the major proteins of intracellular granules and are released with histamine and heparin by activated cells. Most of these proteases are active in the granule as well as outside of the mast cell when secreted, and can cleave targets near degranulating mast cells and in adjoining tissue compartments. Some proteases released from mast cells reach the bloodstream and may have far-reaching actions. In terms of relative amounts, the major mast cell proteases include the tryptases, chymases, cathepsin G, carboxypeptidase A3, dipeptidylpeptidase I/cathepsin C, and cathepsins L and S. Some mast cells also produce granzyme B, plasminogen activators, and matrix metalloproteinases. Tryptases and chymases are almost entirely mast cell-specific, whereas other proteases, such as cathepsins G, C, and L are expressed by a variety of inflammatory cells. Carboxypeptidase A3 expression is a property shared by basophils and mast cells. Other proteases, such as mastins, are largely basophil-specific, although human basophils are protease-deficient compared with their murine counterparts. The major classes of mast cell proteases have been targeted for development of therapeutic inhibitors. Also, a human β-tryptase has been proposed as a potential drug itself, to inactivate of snake venins. Diseases linked to mast cell proteases include allergic diseases, such as asthma, eczema, and anaphylaxis, but also include non-allergic diseases such as inflammatory bowel disease, autoimmune arthritis, atherosclerosis, aortic aneurysms, hypertension, myocardial infarction, heart failure, pulmonary hypertension and scarring diseases of lungs and other organs. In some cases, studies performed in mouse models suggest protective or homeostatic roles for specific proteases (or groups of proteases) in infections by bacteria, worms and other parasites, and even in allergic inflammation. At the same time, a clearer picture has emerged of differences in the

  2. A Disease Marker for Aspirin-Induced Chronic Urticaria

    Directory of Open Access Journals (Sweden)

    Chia-Wei Hsieh

    2014-07-01

    Full Text Available There are currently no diagnostic methods in vitro for aspirin-induced chronic urticaria (AICU except for the provocation test in vivo. To identify disease markers for AICU, we investigated the single nucleotide polymorphism (SNP of the promoter loci of high-affinity IgE receptor (FcεRIα and CD203c expression level in Chinese patients with AICU. We studied two genotypic and allelic frequencies of rs2427827 (–344C/T and rs2251746 (–66T/C gene polymorphisms of FcεRIα in 20 patients with AICU, 52 subjects with airway hypersensitivity without aspirin intolerance, and 50 controls in a Chinese population. The results showed that the frequencies of two SNPs (–344C>T, –66C>T were similar to the normal controls. The allele frequency of –344CC was significantly higher in the patients with AICU compared to those with airway sensitivity (p = 0.019. We also studied both histamine release and CD203c expression on KU812 cells to assess aspirin-induced basophil activation. We found that the activity of basophil activation of AICU was significantly higher in the patients with AICU compared to those with airway hypersensitivity without aspirin intolerance. The mean fluorescence intensity of the CD203c expression were 122.5 ± 5.2 vs. 103.3 ± 3.3 respectively, (p < 0.05, and the percentages of histamine release were 31.3% ± 7.4% vs. −24.0% ± 17.5%, (p < 0.05 respectively. Although the mean fluorescence intensity of CD203c expression and the percentage of histamine release were significantly up-regulated by aspirin, they were not affected by anti-IgE antibodies. These results suggest that a single SNP of FcεRIα (–344C>T is less likely to develop AICU and the basophil activation activity in the sera by measuring CD203c expression can be applicable to confirm the diagnosis of AICU.

  3. Haemopedia: An Expression Atlas of Murine Hematopoietic Cells

    Directory of Open Access Journals (Sweden)

    Carolyn A. de Graaf

    2016-09-01

    Full Text Available Hematopoiesis is a multistage process involving the differentiation of stem and progenitor cells into distinct mature cell lineages. Here we present Haemopedia, an atlas of murine gene-expression data containing 54 hematopoietic cell types, covering all the mature lineages in hematopoiesis. We include rare cell populations such as eosinophils, mast cells, basophils, and megakaryocytes, and a broad collection of progenitor and stem cells. We show that lineage branching and maturation during hematopoiesis can be reconstructed using the expression patterns of small sets of genes. We also have identified genes with enriched expression in each of the mature blood cell lineages, many of which show conserved lineage-enriched expression in human hematopoiesis. We have created an online web portal called Haemosphere to make analyses of Haemopedia and other blood cell transcriptional datasets easier. This resource provides simple tools to interrogate gene-expression-based relationships between hematopoietic cell types and genes of interest.

  4. Anaphylaxis from atracurium without skin manifestation

    Directory of Open Access Journals (Sweden)

    Souvik Maitra

    2014-01-01

    Full Text Available Anaphylaxis is an acute, potentially fatal allergic reaction involving multi organ system that is triggered by a wide range of antigens with a subsequent release of chemical mediators from mast cells and basophils. Diagnosis is primarily clinical though laboratory studies are helpful in further confirmation. Anaphylactic reactions during anesthesia are rare, but can be fatal if not promptly recognized and treated. Among all drugs commonly used in anesthesia, muscle relaxants are the most notorious to trigger anaphylactic reactions and benzylisoquinolinium group of drugs are known to be less common an offender than the steroidal compounds. We report severe anaphylactic reaction after administration of atracurium that was promptly diagnosed and managed without any further morbidity.

  5. Generation, isolation, and maintenance of rodent mast cells and mast cell lines

    DEFF Research Database (Denmark)

    Jensen, Bettina M; Swindle, Emily J; Iwaki, Shoko;

    2006-01-01

    Antigen-mediated mast cell activation, with subsequent mediator release, is a major initiator of the inflammatory allergic response associated with such conditions as asthma. A comprehensive understanding of the principles involved in this process therefore is key to the development of novel...... therapies for the treatment of these disease states. In vitro models of mast cell function have allowed significant progress to be made in the recognition of the fundamental principles of mast cell activation via the high-affinity IgE receptor (FcvarepsilonRI) and, more recently, other receptors expressed...... on mast cells. In addition to human mast cells, the major cell culture systems employed to investigate these responses are rat and mouse peritoneal mast cells, mouse bone-marrow-derived mast cells, the rat basophilic leukemia cell line RBL-2H3, and the mouse MC/9 mast cell line. In this unit, we describe...

  6. Allergen-specific IgE testing in the diagnosis of food allergy and the event of a positive match in the bioinformatics search

    DEFF Research Database (Denmark)

    van Ree, Ronald; Vieths, Stefan; Poulsen, Lars K.

    2006-01-01

    Current documents on risk assessment of genetically modified foods recommend including IgE-binding tests on sera from allergic patients. However, there is no generally accepted recommendation on technical aspects of the testing procedures or on the interpretation of the results, despite that fact...... that the use of sera from clinically well-characterized subjects is of high importance. In the case of a positive test result, the biological activity of the detected IgE antibodies, i. e., the potential to trigger mediator release from basophils or mast cells in an allergen-specific manner, should be taken...... screening, i. e., the testing of gene products from organisms not known to be allergenic with sera from subjects allergic to related species....

  7. The prevalence of suspected and challenge-verified penicillin allergy in a university hospital population

    DEFF Research Database (Denmark)

    Borch, Jacob Eli; Andersen, Klaus Ejner; Bindslev-Jensen, Carsten

    2006-01-01

    Suspected penicillin allergy is common among hospitalised patients, but the quality of the information given by the patient is often doubtful. Alleged penicillin allergic are likely to be treated with more toxic, broad-spectrum, and more expensive antibiotics, with effects on microbial resistance...... patterns and public economy as a consequence. We performed a cross-sectional case-control study with two visits to all clinical departments of a large university hospital in order to find in-patients with medical files labelled "penicillin allergy" or who reported penicillin allergy upon admission. Patient...... histories were obtained via a questionnaire, and they were offered investigation for penicillin allergy with specific IgE, basophil histamine release, skin prick tests, intradermal tests and drug challenge tests. Finally, the pharmaco-economical consequences of the penicillin allergy were estimated...

  8. The use of ciclosporin A in veterinary dermatology.

    Science.gov (United States)

    Kovalik, Marcel; Thoday, Keith L; van den Broek, Adri H M

    2012-08-01

    Ciclosporin A (CsA) has potent immunosuppressive and immunomodulatory activity that has been exploited in human medicine to prevent the rejection of transplanted organs and to manage atopic dermatitis and psoriasis. Over the past decade, CsA has been employed more frequently in veterinary dermatology and its value in the management of several canine and feline dermatoses is now well established. CsA inhibits calcineurin phosphatase, suppressing T cell activation and the synthesis of T cell cytokines consequently impairing the activity of B cells, antigen-presenting cells, mast cells, basophils and eosinophils. The pharmacokinetics of CsA are similar in humans, dogs and cats and the drug has a wide safety margin in dogs, cats and rabbits. Adverse effects, principally transient vomiting and soft faeces/diarrhoea, may be seen shortly after instituting treatment but often resolve despite continuing treatment. Gingival hyperplasia and cutaneous effects such as hirsutism may occur after prolonged treatment.

  9. [Is Malherbe calcifying epithelioma in the ENT area of a rare tumor?].

    Science.gov (United States)

    Ganz, H

    1986-07-01

    The calcifying epithelioma (pilomatrixoma) is a benign tumour of the skin originating from hair sheath cells. The head, especially the cheek and preauricular region with the parotid gland, is a very common site (about 50%). The tumour arises predominantly in younger people, with a 2:1 female to male ratio. It is not so rare as suggested by the few ENT publications. The histologic picture shows two characteristic epithelial cells, the basophilic and ghost cell. Calcification is present in most cases. The prognosis after removal of the tumour is good. Two personal cases are reported, multiple pilomatrixomas on the cheek of an 12 year old girl, and a tumour on the auricle, an unusual site.

  10. Healing of bone in the rat following surgery with the erbium-YAG laser

    Science.gov (United States)

    Dickinson, Mark R.; Devlin, Hugh; El Montaser, Monsour A.; Sloan, Philip

    1996-12-01

    Background and objectives: the aim of this study was to examine the pattern of healing in rat calvarial defects prepared with the erbium-YAG laser, using the 'guided tissue regeneration' technique. Materials and method: PTFE membranes were placed over lased skull defects, and the skin wounds sutured. Rats were killed humanely at intervals after surgery, and the skulls processed for paraffin wax histology. A further group of mature rats were also killed humanely and the calvariae removed. Slots were prepared using the erbium-YAG laser and immediately examined under the environmental scanning electron microscope (ESEM) in hydrated conditions, which avoided drying artifacts. Results: An amorphous, mineral-rich carbon layer surrounds the lased bone defect, which in the in vivo experiments was seen as a basophilic zone which was resistant to resorption.

  11. Candidatus Renichlamydia lutjani, a Gram-negative bacterium in internal organs of blue striped snapper Lutjanus kasmira from Hawaii.

    Science.gov (United States)

    Corsaro, Daniele; Work, Thierry M.

    2012-01-01

    The blue-striped snapper Lutjanus kasmira (Perciformes, Lutjanidae) are cosmopolitan in the Indo-Pacific but were introduced into Oahu, Hawaii, USA, in the 1950s and have since colonized most of the archipelago. Studies of microparasites in blue-striped snappers from Hawaii revealed chlamydia-like organisms (CLO) infecting the spleen and kidney, characterized by intracellular basophilic granular inclusions containing Gram-negative and Gimenez-positive bacteria similar in appearance to epitheliocysts when seen under light microscopy. We provide molecular evidence that CLO are a new member of Chlamydiae, i.e. Candidatus Renichlamydia lutjani, that represents the first reported case of chlamydial infection in organs other than the gill in fishes.

  12. Detection and significance of neuropeptide Y and CD63 in patients with urticaria factitia%人工荨麻疹患者神经肽Y和CD63的检测及意义

    Institute of Scientific and Technical Information of China (English)

    王爱学; 李玉平; 王瑞敏; 卢凤敏; 郭兵申; 张环环; 石丽萍; 姚丽美; 四荣联

    2013-01-01

    Objective To investigate the role of neuropeptide Y and activated basophilic granulocytes in the pathogenesis of urticaria factitia.Methods This study included 24 outpatients with urticaria factitia collected from August 2011 to December 2011 at the Department of Dermatology,Second Hospital of Hebei Medical University,as well as 16 healthy volunteers without allergic diseases,chronic somatic diseases,nervous system disorders or psychiatric disorders.Venous blood samples were collected from these subjects.Radioimmunoassay was carried out to determine the plasma level of neuropeptide Y,and flow cytometric analysis to count activated basophilic granulocytes after triple staining with CD45-peridinin-chlorophyll-protein complex/CD203c-phycoerythrin/CD63-fluorescein isothiocyanate.Results The plasma level of neuropeptide Y was (111.155 ±36.832) ng/L and (104.456 ± 44.697) ng/L in the patients and healthy volunteers respectively (t =0.198,P >0.05).The median number of activated basophilic granulocytes was significantly higher in the patients than in the healthy controls (280.5 × 106/L vs.18.1 × 106/L,P <0.05).Conclusisons Activated basophilic granulocytes appear to be involved in the pathogenesis of urticaria factitia,while the role of neuropeptide Y remains unknown.%目的 探讨神经肽Y和活化的嗜碱性粒细胞在人工荨麻疹发病机制中的意义.方法 人工荨麻疹患者24例来自2011年8-12月河北医科大学第二医院皮肤科门诊,16例对照为除外过敏性疾病、躯体慢性疾病、神经系统及精神疾病的同期健康志愿者.用放射免疫分析法检测血浆神经肽Y的表达;用CD45-PERCP(多甲藻黄素-叶绿素-蛋白质复合物)、CD203c-藻红蛋白、CD63-异硫氰酸荧光素三联抗体标记嗜碱性粒细胞,流式细胞仪检测CD63的表达水平,计数活化嗜碱性粒细胞数量.结果 人工荨麻疹患者与健康对照组神经肽Y的含量分别为(111.155±36.832) ng/L和(104.456±44.697) ng/L

  13. Ara h 6 complements Ara h 2 as an important marker for IgE reactivity to peanut.

    Science.gov (United States)

    Koid, Audrey E; Chapman, Martin D; Hamilton, Robert G; van Ree, Ronald; Versteeg, Serge A; Dreskin, Stephen C; Koppelman, Stef J; Wünschmann, Sabina

    2014-01-08

    The similarities of two major peanut allergens, Ara h 2 and Ara h 6, in molecular size, amino acid sequence, and structure have made it difficult to obtain natural Ara h 6 free of Ara h 2. The objectives of this study were to purify natural Ara h 6 that is essentially free of Ara h 2 and to compare its IgE reactivity and potency in histamine release assays to Ara h 2. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the highly purified allergen (basophil histamine release assays than Ara h 2. Purified Ara h 6 will be useful for diagnostic IgE antibody assays as well as molecular and cellular studies to investigate the immunological mechanisms of peanut allergy.

  14. DSCG binding protein and process for preparing same

    Energy Technology Data Exchange (ETDEWEB)

    Pecht, I.; Mazurek, N.

    1987-07-28

    An essentially pure protein is described consisting essentially of the protein, (CBP), present in nature in membranes of basophile cells and in mast cells, having a molecular weight of about 60,000 +- 2,000 determined by SDS polyacrylamide electrophoresis an isoelectric point of about 3.9 and an amino acid composition of about 4 units of asparagine, 3 units of threonine and serine, 3 units glycine, 2 units alanine, 2 units proline, 1 unit cysteine, 2 units valine, 1 unit methionine, 1 unit isoleucine, 2 units leucine, 1 unit tyrosine, 1 unit phenylalanine, 2 units histamine, 2 units lysine and 1 unit arginine. The protein is able to build calcium and having a calcium dependent affinity to the disodium salt of 1,2 bis(-2 carboxychromon-5-yloxy)-2-hydroxy propane (DSCG).

  15. The second PGD(2) receptor CRTH2: structure, properties, and functions in leukocytes.

    Science.gov (United States)

    Nagata, Kinya; Hirai, Hiroyuki

    2003-01-01

    Prostaglandin (PG) D(2) plays a broad range of physiological and pathophysiological functions. Until just a few years ago, it was thought that most of the biological actions of PGD(2) are mediated via the classical PGD(2) receptor DP. Recently, we identified a second PGD(2) receptor, chemoattractant receptor-homologous molecule expressed on T helper (Th)2 cells (CRTH2), with different functions relative to DP. Here, we review the recent findings on the structure, tissue distribution, ligand selectivity, signalling pathways, and functions in leukocytes of this receptor. The data suggest that the PGD(2)/CRTH2 system play important roles in allergic inflammation through its stimulatory effects on Th2 cells, eosinophils, and basophils.

  16. Effects of cellular fine structure on scattered light pattern.

    Science.gov (United States)

    Liu, Caigen; Capjack, Clarence E

    2006-06-01

    Biological cells are complex in both morphological and biochemical structure. The effects of cellular fine structure on light scattered from cells are studied by employing a three-dimensional code named AETHER which solves the full set of Maxwell equations by using the finite-difference time-domain method. It is shown that changes in cellular fine structure can cause significant changes in the scattered light pattern over particular scattering angles. These changes potentially provide the possibility for distinguishability of cellular intrastructures. The effects that features of different intrastructure have on scattered light are discussed from the viewpoint of diagnosing cellular fine structure. Finally, we discuss scattered light patterns for lymphocyte-like cells and basophil-like cells.

  17. Regulation of hematopoietic cell function by inhibitory immunoglobulin G receptors and their inositol lipid phosphatase effectors.

    Science.gov (United States)

    Cady, Carol T; Rice, Jeffrey S; Ott, Vanessa L; Cambier, John C

    2008-08-01

    Numerous autoimmune and inflammatory disorders stem from the dysregulation of hematopoietic cell activation. The activity of inositol lipid and protein tyrosine phosphatases, and the receptors that recruit them, is critical for prevention of these disorders. Balanced signaling by inhibitory and activating receptors is now recognized to be an important factor in tuning cell function and inflammatory potential. In this review, we provide an overview of current knowledge of membrane proximal events in signaling by inhibitory/regulatory receptors focusing on structural and functional characteristics of receptors and their effectors Src homology 2 (SH2) domain-containing tyrosine phosphatase 1 and SH2 domain-containing inositol 5-phosphatase-1. We review use of new strategies to identify novel regulatory receptors and effectors. Finally, we discuss complementary actions of paired inhibitory and activating receptors, using Fc gammaRIIA and Fc gammaRIIB regulation human basophil activation as a prototype.

  18. Structure of capsule around acanthocephalan Corynosoma strumosum from uncommon paratenic hosts-lizards of two species.

    Science.gov (United States)

    Skorobrechova, Ekaterina M; Nikishin, Vladimir P; Lisitsyna, Olga I

    2012-01-01

    Micromorphology and ultrastructure of capsule forming around acanthocephalan Corynosoma strumosum in uncommon paratenic hosts-lizards Lacerta agilis and Lacerta viridis-have been studied. Experimental infestation of the lizards by acanthocephalans obtained from naturally infested sea fishes showed that only small amount of parasites occurred in the intestine of the host was able to migrate into body cavity and to be encapsulated. Micromorphology of capsules of different ages from different species of lizards and micromorphology and ultrastructure of capsules at the age of 1.5 and 10 days appeared to be similar. In the capsule's structure cells of inflammatory rank were prevailing: mononuclear and multinuclear macrophages, eosinophils, and basophils. Fibroblasts were not numerous and were located only in the outer part of a capsule; exocellular collagen fibers were absent. Inflammatory character of capsule confirms the idea that lizards are unsuitable paratenic hosts for corynosomes.

  19. Tomato ( Lycopersicon esculentum ) seeds: new flavonols and cytotoxic effect.

    Science.gov (United States)

    Ferreres, Federico; Taveira, Marcos; Pereira, David M; Valentão, Patrícia; Andrade, Paula B

    2010-03-10

    In this study, seeds of Lycopersicon esculentum Mill. were analyzed by HPLC/UV-PAD/MS(n)-ESI. Fourteen flavonoids were identified, including quercetin, kaempferol, and isorhamnetin derivatives, with 13 of them being reported for the first time in tomato seeds. The major identified compounds were quercetin-3-O-sophoroside, kaempferol-3-O-sophoroside, and isorhamnetin-3-O-sophoroside. A significant cell proliferation inhibition (>80%), against rat basophile leukemia (RBL-2H3) cell line, was observed with this extract (IC(50) = 5980 microg/mL). For acetylcholinesterase inhibitory activity, a concentration-dependent effect was verified (IC(20) = 2400 microg/mL). The same behavior was noted regarding antioxidant capacity, evaluated against DPPH (IC(10) = 284 microg/mL), nitric oxide (IC(25) = 396 microg/L), and superoxide radicals (IC(25) = 3 microg/mL).

  20. Lead Toxicity Resulting from Chronic Ingestion of Opium

    Directory of Open Access Journals (Sweden)

    Jalili, Mohammad

    2009-11-01

    Full Text Available A 32-year-old man presented to the emergency department (ED with lower abdominal pain and constipation. He related chronic ingestion of large amounts of opium. Physical examination showed mild abdominal tenderness and gingival discoloration. Diagnostic studies showed a mild hypochromic, microcytic anemia with basophilic stippling of the red blood cells. Abdominal imaging showed no intra-abdominal pathology. A diagnosis of lead toxicity was confirmed through serum lead levels. The patient was put on chelation therapy and his signs and symptoms started to resolve. As a comprehensive search for other sources of lead was unsuccessful, opium adulterants were considered as the culprit. Chemical analysis of the opium confirmed this. Contaminated drugs have been reported as a source of exposure to toxins such as arsenic or lead. While other reports deal with patients from clinics, this report illustrates lead toxicity from ingestion of contaminated opium in the ED.[West J Emerg Med. 2009;10(4:244-246.

  1. Correlated TEM, SEM, and histological observations of filiform papillae of the cow tongue.

    Science.gov (United States)

    Steflik, D E; Singh, B B; Mckinney, R V; Boshell, J L

    1983-01-01

    The ultrastructural and histological characteristics of the filiform papillae of the cow tongue are described. These large, conical papillae display a fully keratinized external projection, which demonstrates a dominant hard keratin core partially ensheathed by soft keratin. This biostructure differentiates from a dominant posterior cell line, leading to the hard keratin core, and a thinner anterior cell line which, with contributions from an interpapillary cell line, appears responsible for an attenuating sheath of soft keratin around the hard keratin core. The hard keratin cell line differentiates from cells devoid of keratohyalin granules (KHG), which are rich in thick bundles of tonofilaments. The soft keratin cell line differentiates from cells containing both eosinophil and basophil KHG and dispersed tonofilaments. The bovine filiform papilla also appears similar in biostructure to the mammalian hair shaft and the 'filiform hairs' of the rat penis.

  2. IL-10-Producing Type 1 Regulatory T Cells and Allergy

    Institute of Scientific and Technical Information of China (English)

    Kui Wu; Yutian Bi; Kun Sun; Changzheng Wang

    2007-01-01

    As an important subset of regulatory T (Treg) cells, IL-10-producing type 1 regulatory T cells (Tr1), have some different features to thymic-derived naturally occurring CD4+CD25+Foxp3+ Treg cells(nTreg cells). Similar to nTreg cells, Tr1 also play important roles in the control of allergic inflammation in several ways. There is a fine balance between Tr1 and Th2 responses in healthy subjects. Skewing of allergic-specific effctor T cells to a Tr1 phenotype appears to be a critical event in successful allergen-specific immunotherapy and glucocorticoids and β2-agonists treatment. Tr1 suppress Th2 cells and effector cells of allergic inflammation, such as eosinophils, mast cells, basophils, through producing IL-10, and perhaps TGF-β. Understanding of Tr1 may be helpful in developing new strategies for treatment of allergic diseases.

  3. Bacteria-induced histamine release from human bronchoalveolar cells and blood leukocytes

    DEFF Research Database (Denmark)

    Clementsen, P; Milman, N; Struve-Christensen, E

    1991-01-01

    Histamine release induced by Staphylococcus aureus was examined in cells obtained by bronchoalveolar lavage (BAL) in non-atopic individuals. Approximately half of the individuals responded with mediator release to the bacterium, and the release was found to be time- and concentration dependent....... No difference was found between the patients who responded and those who did not respond in regard to age, sex, smoker/non-smoker, % recovery of BAL-fluid, total cell count, differential cell counts, histamine content per mast cell, or diagnoses. Also stimulation of the BAL-cells with the calcium-ionophore A......23187 resulted in histamine release. S. aureus-induced histamine release from basophils was examined in leukocyte suspensions obtained from the same individuals, and in all experiments release was found. The dose-response curves were similar to those obtained with BAL cells. The bacteria...

  4. Accelerated phase chronic myeloid leukemia: evaluation of clinical criteria as predictors of survival, major cytogenetic response and progression to blast phase

    Directory of Open Access Journals (Sweden)

    Vanessa Fiorini Furtado

    2015-10-01

    Full Text Available BACKGROUND: Published criteria defining the accelerated phase in chronic myeloid leukemia are heterogeneous and little is known about predictors of poor outcome.METHODS: This is a retrospective study of 139 subjects in the accelerated phase of chronic myeloid leukemia treated with imatinib at a single center in Brazil. The objective was to identify risk factors for survival, major cytogenetic response and progression to blast phase in this population. The factors analyzed were: blasts 10-29%, basophils ≥ 20%, platelets > 1 × 106/µL or 1 × 105/µL in the peripheral blood, as well as clonal evolution, splenomegaly, hemoglobin 12 months (p-value = 0.030.CONCLUSION: These data indicate that patients with the above risk factors have a worse prognosis. This information can guide the therapy to be used.

  5. Haemato-biochemical alterations induced by lead acetate toxicity in wistar rats

    Directory of Open Access Journals (Sweden)

    S. G. Suradkar

    Full Text Available An experiment was conducted to study the haemato-biochemical alterations induced by lead acetate toxicity in 48 Wistar rats of either sex, divided uniformly into four different groups. The rats of group I received only deionised water as control while, group II, III and IV were given lead acetate @ 1 PPM, 100 PPM and 1000 PPM, in drinking deionised water respectively for 28 days. In group III and IV dose dependant significant (P<0.05 reductions in TEC, Hb, PCV and TLC were observed. No significant change was observed in neutrophil, eosinophil, basophil and monocyte count in any treatment groups, whereas the lymphocyte count decreased significantly (P<0.05 in group III and IV. A dose dependant significant (P<0.05 increase in AST, ALP, AKP, GGT, BUN and creatinine was experiential while TP and albumin levels were decreased in group III and IV. [Vet World 2009; 2(11.000: 429-431

  6. Ileum and colon perforation following peritoneal dialysis-related peritonitis and high-dose calcium polystyrene sulfonate.

    Science.gov (United States)

    Kao, Chih-Chin; Tsai, Yi-Chiun; Chiang, Wen-Chih; Mao, Tsui-Lien; Kao, Tze-Wah

    2015-10-01

    A rare but severe complication, intestinal necrosis, has been reported after sodium polystyrene sulfonate (SPS; Kayexalate) and sorbitol intake. Some case reports described bowel perforation following calcium polystyrene sulfonate (CPS; Kalimate) administration. We report a case of ileum and colon perforation following peritoneal dialysis-related peritonitis and high-dose Kalimate in a 59-year-old female patient. The patient had a history of hypertension, diabetes mellitus, and end-stage renal disease (ESRD). During hospitalization for peritoneal dialysis-related peritonitis, she developed hyperkalemia, and Kalimate was administered orally. However, severe abdominal distension and pain occurred just one day after Kalimate intake. An urgent surgery disclosed several perforations in the ileum and sigmoid colon. Pathology of the resected gut showed transmural necrosis and perforation with basophilic angulated crystals. The patient finally expired during hospitalization due to refractory septic shock.

  7. Traditional medicine: a rare cause of lead poisoning in Western countries [v1; ref status: indexed, http://f1000r.es/2c6

    Directory of Open Access Journals (Sweden)

    Halima Muller

    2013-11-01

    Full Text Available A 42-year-old man from Bhutan was admitted to the emergency department with a 5-day history of abdominal pain, nausea and vomiting. Enhanced abdominal CT scan was found negative, however laboratory tests showed hemolytic anemia and basophilic stippling which are often seen in lead and heavy metal poisoning. Additional tests revealed a high level of lead in blood and urine. The patient was administered a chelator treatment with rapid improvement of the symptoms. A detailed interview revealed that the patient had been taking daily Bhutanese traditional medicines to treat a Bell’s palsy from which he had been suffering for a few months. The analysis of these medicines confirmed the presence of a high level of lead.

  8. A case of taurine-containing drink induced anaphylaxis.

    Science.gov (United States)

    Lee, Seung-Eun; Lee, Suh-Young; Jo, Eun-Jung; Kim, Mi-Young; Yang, Min-Suk; Chang, Yoon-Seok; Kim, Sae-Hoon

    2013-01-01

    Taurine is one of most abundant free amino acids in mammalian tissue. It has been used for various health functional foods as a main ingredient in food industry. A 33-year-old female patient repeatedly experienced generalized itching, urticaria, dyspnea and dizziness after drinking taurine-containing drinks. The patient showed positive response to oral challenge tests with taurine-containing drinks. The patient also showed positive response with synthetic taurine but not with natural taurine. Skin prick test and basophil activation test with the synthetic taurine were negative. To our knowledge, there has been no report of taurine-induced hypersensitivity reactions. We herein report the first case of taurine-containing drink induced anaphylaxis, especially by synthetic taurine.

  9. Cutaneous hybrid tumor composed of epidermal cyst and cystic pilomatricoma expressing p53 and high Ki-67 labeling.

    Science.gov (United States)

    Terada, Tadashi

    2013-01-01

    Hybrid tumor composed of epidermal cyst and pilomatricoma has been reported in only four times in the English literature. Herein, a cutaneous hybrid tumor composed of epidermal cyst and cystic pilomatricoma in a 58-year-old woman was presented. The tumor was located in the scalp, and measured 1 x 1 x 1 cm. The tumor was cystic and contained atheromatous materials. Histologically, the cyst was composed of epidermal cyst (50% in area) and pilomatricoma (50% in area). The pilomatricoma was composed mostly of basophilic cells. Interestingly, the pilomatricoma element of the cyst showed immunoreactive p53 and Ki-67 (labeling=40%). In conclusion, the fifth case of hybrid tumor composed of epidermal cyst and pilomatricoma was presented.

  10. Outbreak of herpesviral conjunctivitis and respiratory disease in gouldian finches.

    Science.gov (United States)

    Paulman, A; Lichtensteiger, C A; Kohrt, L J

    2006-11-01

    An outbreak of tracheitis, sinusitis, and conjunctivitis, originating in recently imported birds, caused high morbidity and mortality in a flock of finches in Central Illinois. Although several species were present, Gouldian finches (Erythrura [Chloebia] gouldiae) were most commonly and severely affected. Birds submitted for necropsy displayed microscopic lesions characteristic of herpesviral infection, including epithelial cytomegaly and karyomegaly with basophilic, intranuclear inclusion bodies in the nasopharynx, sinuses, trachea, parabronchi, conjunctiva, and occasionally the lacrimal gland or proximal proventricular glands. Viral particles consistent with herpesvirus were visualized within affected epithelial cells with electron microscopy. Based on a partial sequence of the viral DNA polymerase gene, this virus was found to be identical to a herpesvirus previously implicated in a similar outbreak in Canada and is most likely an alphaherpesvirus.

  11. Identification and characterization of a novel adenovirus in the cloacal bursa of gulls.

    Science.gov (United States)

    Bodewes, R; van de Bildt, M W G; Schapendonk, C M E; van Leeuwen, M; van Boheemen, S; de Jong, A A W; Osterhaus, A D M E; Smits, S L; Kuiken, T

    2013-05-25

    Several viruses of the family of Adenoviridae are associated with disease in birds. Here we report the detection of a novel adenovirus in the cloacal bursa of herring gulls (Larus argentatus) and lesser black-backed gulls (Larus fuscus) that were found dead in the Netherlands in 2001. Histopathological analysis of the cloacal bursa revealed cytomegaly and karyomegaly with basophilic intranuclear inclusions typical for adenovirus infection. The presence of an adenovirus was confirmed by electron microscopy. By random PCR in combination with deep sequencing, sequences were detected that had the best hit with known adenoviruses. Phylogenetic analysis of complete coding sequences of the hexon, penton and polymerase genes indicates that this novel virus, tentatively named Gull adenovirus, belongs to the genus Aviadenovirus. The present study demonstrates that birds of the Laridae family are infected by family-specific adenoviruses that differ from known adenoviruses in other bird species.

  12. Blood cell profiles of the tadpoles of the Dubois's tree frog, Polypedates teraiensis Dubois, 1986 (Anura: Rhacophoridae).

    Science.gov (United States)

    Das, Madhusmita; Mahapatra, Pravati Kumari

    2012-01-01

    The present paper describes a sequential study of the leukocyte profiles and the changes in morphometry and morphology of erythrocytes in the tadpoles of Polypedates teraiensis during their development and metamorphosis, that is, transfer from an aquatic mode to a terrestrial mode of life. Blood smears of 21 different stages (Gosner stage 26 to 46) of tadpoles were investigated. Population of erythrocytes was heterogeneous in population represented by various forms (oval, elliptical or rounded cells, comma shaped, teardrop shaped, schistocytes, senile erythrocytes, crenulated RBCs). Correlation between various morphometric values of erythrocytes was determined with different developing stages of tadpoles. Amongst the leucocytes, the lymphocytes were the most abundant cells followed by neutrophils. Neutrophils and monocytes showed varied morphologic forms. The percentage of lymphocytes and neutrophils showed a negative whereas percentage of eosinophil, basophil, and monocytes showed a positive correlation with the developmental stages of tadpoles. Blood platelets were also observed, which were rounded in shape and found in aggregates.

  13. Using bacteria to determine protein kinase specificity and predict target substrates.

    Directory of Open Access Journals (Sweden)

    Michael F Chou

    Full Text Available The identification of protein kinase targets remains a significant bottleneck for our understanding of signal transduction in normal and diseased cellular states. Kinases recognize their substrates in part through sequence motifs on substrate proteins, which, to date, have most effectively been elucidated using combinatorial peptide library approaches. Here, we present and demonstrate the ProPeL method for easy and accurate discovery of kinase specificity motifs through the use of native bacterial proteomes that serve as in vivo libraries for thousands of simultaneous phosphorylation reactions. Using recombinant kinases expressed in E. coli followed by mass spectrometry, the approach accurately recapitulated the well-established motif preferences of human basophilic (Protein Kinase A and acidophilic (Casein Kinase II kinases. These motifs, derived for PKA and CK II using only bacterial sequence data, were then further validated by utilizing them in conjunction with the scan-x software program to computationally predict known human phosphorylation sites with high confidence.

  14. NF-κB Regulates B-Cell-Derived Nerve Growth Factor Expression

    Institute of Scientific and Technical Information of China (English)

    Klaus Heese; Noriko Inoue; Tohru Sawada

    2006-01-01

    In the mammalian brain, four neurotrophins have been identified: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5). NGF exerts an important role in the development and functions of the central and peripheral nervous system. However, it has recently been documented that several types of immune cells, such as mast cells, lymphocytes, basophils and eosinophils, produce,store and release NGF. Accumulating preclinical and clinical data indicate that dysfunctions of NGF and the other neurotrophins may contribute to impaired immune responses and concentration of NGF frequently correlates with disease severity. Thus, the aim of this study was to elucidate the potential signaling mechanisms of cytokineneurotrophins interactions contributing to increased NGF levels. Our data show that the transcription factorNF-κB plays a pivotal role in regulating B-cell-derived NGF expression.

  15. Histopathological Observation of Lymphocystis Disease and Lymphocystis Disease Virus (LCDV) Detection in Cultured Diseased Sebastes schlegeli

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Lymphocystis nodules occurring in the cultured sting fish Sebastes schlegeli were observed under light and electron microscope. Lymphocystis disease virus (LCDV) in the tissues of diseased fish was detected with indirect immunofluorescence test (IFAT). Results showed that lymphocystis cells had overly irregular nuclei, basophilic intracytoplasmic inclusion bodies with virions budding from the surface, and hyaline capsules outside the cell membrane. Numerous virus particles about 200 nm in diameter scattered in the cytoplasm, electron-dense particles 70-80nm in diameter filled in perinuclear cisterna, and membrane-enveloped particles with electron-dense core of 70-80 nm appeared around cellular nucleus. IFAT using monoclonal antibody against LCDV from Paralichthys olivaceus revealed that specific green fluorescence was present in the cytoplasm of lymphocystis cells, epithelium of stomach, gill lamellae, and muscular fibers under epidermis of S. schlegeli, just as that in the cytoplasm of lymphocystis cells of P. olivaceus, suggesting the presence of LCDV in these tissues.

  16. Granulocyte and granulocyte macrophage colony-stimulating factors as therapy in human and veterinary medicine.

    Science.gov (United States)

    Fernández-Varón, Emilio; Villamayor, Lucía

    2007-07-01

    Granulocyte colony-stimulating factors (G-CSFs) and granulocyte macrophage colony-stimulating factors (GM-CSFs) are endogenous cytokines that regulate granulocyte colonies and play a major role in the stimulation of granulopoiesis (neutrophils, basophils and eosinophils) and in the regulation of microbicidal functions. There are numerous pathological conditions in which neutrophils are decreased, the most common being neutropenia associated with cancer chemotherapy, which increases the risk of serious microbial infections developing with the potential for high morbidity and mortality. New methods in molecular biology have led to the identification and cloning of CSF genes and biopharmaceutical production. Since then, CSFs have been widely used for the prevention and treatment of neutropenia associated with cancer chemotherapy, for mobilising haematopoietic cell precursors, and for other neutropenia-related pathologies. This review focuses on the use of CSFs within both human and veterinary medicine. Clinical applications, pharmacology, tolerability and the potential role of these factors in veterinary medicine are considered.

  17. [Differential analysis of BM cell morphology, immunophenotypic, cytogenetic characters and prognosis between myeloblastic and lymphoblastic crisis of CML].

    Science.gov (United States)

    Liu, Ya-Lin; Wang, Xiao-Ning; Liu, Hua-Sheng

    2014-06-01

    This study was purposed to investigate the difference of morphology, immunophenotype, cytogenetic features and prognosis between myeloid blast crisis and lymphoid blast crisis of chronic myelogenous leukemia (CML). A total of 31 patients with CML in blastic crisis in Department of Hematology, the First Affiliated Hospital of Xi'an Jiaotong University school of Medicine from 2009 January to 2014 January were enrolled in this study. Out of 31 CML patients, 24 cases were patients with myeloid blast crisis and other 7 cases were patients with lymphoblastic crisis. The clinical data, blast cell percentage in peripheral blood and bone marrow, eosinophil and basophil percentage, immunophenotype, cytogenetic characteristics and prognosis were analyzed. The results indicated that there was no significant difference of blastic cell percentage in peripheral blood and bone marrow of CML with myeloid blast crisis, and the eosinophil and basophil cells could be easily detected. The ratio of blastic cells in BM was higher than that in PB in lymphoid blastic crisis of CML, eosinophil and basophil cells were rare. 7 cases of CML with lymphoid blastic crisis were B ALL with CD10, CD19, CD34, HLA-DR expression, and 2 cases with CD13 and CD33 expression. The lymphoid score was in all CML patients with lymphoid blastic crisis was greater than or equal to 1.5;and 2 patients with CD13 and CD33 expression, and with 1 myeloid score.24 cases of myeloid blastic crisis of CML patients mainly expressed CD33, CD13, CD38, CD34, CD11b and HLA-DR, and their myeloid score greater than or equal to 2, among them the lymphoid scores of 2 patients were 0.5 and 1 score, respectively. All the 31 patients showed 100% Ph(+) chromosome, among them 3 cases also showed other new chromosome aberrations. There was no significant difference of overall survival rate between lymphoid and myeloid blastic crisis of CML, but the overall survival rate of patients treated with tyrosine kinase inhibitor (TKI ) was

  18. Mechanisms of immune tolerance to allergens: role of IL-10 and Tregs

    Science.gov (United States)

    Akdis, Cezmi A.; Akdis, Mübeccel

    2014-01-01

    During the past 20 years, major advances have been made in understanding the molecular and cellular mechanisms of allergen tolerance in humans. The demonstration of T cell tolerance, particularly that mediated by the immune-suppressive functions of IL-10, led to a major conceptual change in this area. Currently, the known essential components of allergen tolerance include the induction of allergen-specific regulatory subsets of T and B cells, the immune-suppressive function of secreted factors, such as IL-10 and TGF-β, the production of IgG4 isotype allergen–specific blocking antibodies, and decreased allergic inflammatory responses by mast cells, basophils, and eosinophils in inflamed tissues. PMID:25365074

  19. Immunology in the Clinic Review Series; focus on allergies: immunotherapy for food allergy.

    Science.gov (United States)

    Mousallem, T; Burks, A W

    2012-01-01

    There is no approved therapy for food allergy. The current standard of care is elimination of the triggering food from the diet and accessibility to epinephrine. Immunotherapy is a promising treatment approach. While desensitization to most foods seems feasible, it remains unclear if a permanent state of tolerance is achievable. The research team at Duke is pioneering immunotherapy for food allergies. Work here has evolved over time from small open-label pilot studies to larger randomized designs. Our data show that immunological changes associated with immunotherapy include reduction in mast cell reactivity, decreased basophil responses, decreased specific-immunoglobulin (Ig)E, increased IgG4 and induction of regulatory T cells. Immunotherapy has generated much excitement in the food allergy community; however, further studies are needed before it is ready for clinical use.

  20. Systemic herpesvirus and morbillivirus co-infection in a striped dolphin (Stenella coeruleoalba).

    Science.gov (United States)

    Soto, S; González, B; Willoughby, K; Maley, M; Olvera, A; Kennedy, S; Marco, A; Domingo, M

    2012-01-01

    During 2007 a dolphin morbillivirus epizootic affected the western Mediterranean and several striped dolphins (Stenella coeruleoalba) stranded on the Catalonian coasts. One of those animals had severe lymphoid depletion, necrosis and syncytial formation in lymph nodes and spleen, with large basophilic nuclear inclusions compatible with herpesvirus detected by immunohistochemical and ultrastructural examination. Non-suppurative encephalitis with associated morbillivirus antigen and morbillivirus antigen within alveolar macrophages were also observed. A pan-herpesvirus nested polymerase chain reaction amplified a sequence virtually identical to two cetacean herpesvirus sequences previously identified in systemic infections in an Atlantic Cuvier's beaked whale (Ziphius cavirostris) and in a Mediterranean striped dolphin. The herpesviral infection was probably secondary to the immunosuppression caused by the morbillivirus. To our knowledge, this is the first report of a cetacean co-infected by dolphin morbillivirus and herpesvirus with evidence of lesions attributable to both viruses.

  1. Lactoferrin, myeloperoxidase, lysozyme and eosinophil cationic protein in exudate in delayed type hypersensitivity

    DEFF Research Database (Denmark)

    Lerche, A; Bisgaard, H; Christensen, J D;

    1988-01-01

    A skin window technique was used to study the morphology of leucocytes in upper dermis and exudate during nickel challenge in patients with contact allergy to nickel. Contact allergic patients and healthy volunteers tested with a skin widow without addition of nickel to the chamber medium served...... as controls. The morphology of the leucocytes in dermis was studied in biopsies taken 8, 24, or 48 h after skin window application, and in a parallel test the morphology of the exudate was examined by sequential collection of the chamber medium during a 48 h period. The infiltrate in dermis of contact...... in contact allergic patients exposed to nickel showed a dominance of polymorphonuclear granulocytes throughout the study period, while mononuclear cells, eosinophils and basophils were detected at a much lower quantity and with a considerable delay. Further, we studied the kinetics of the leucocyte granule...

  2. The high-affinity immunoglobulin E receptor as pharmacological target.

    Science.gov (United States)

    Blank, Ulrich; Charles, Nicolas; Benhamou, Marc

    2016-05-05

    The high-affinity receptor for immunoglobulin E is expressed mainly on mast cells and basophils, but also on neutrophils, eosinophils, platelets, monocytes, Langerhans and dendritic cells, airway smooth muscle cells and some nerve cells. Its main function is, upon its engagement by IgE and specific antigen, to trigger a powerful defense against invading pathogens and a rapid neutralization of dangerous toxic substances introduced in the body. This powerful response could be wielded against tumors. But, when control over this receptor is lost, its unchecked activation can induce an array of diseases, some of which can lead to death. In this review we will summarize the pharmacological approaches and strategies that are currently used, or under study, to harness or wield activation of this receptor for therapeutic purposes.

  3. Harvey Cushing: Cushing's disease.

    Science.gov (United States)

    Ellis, Harold

    2012-09-01

    In 1932 Harvey Cushing published an article entitled 'Basophil adenomas of the pituitary and their clinical manifestations'. These are rare tumours of the anterior pituitary gland, at the base of the brain, with striking manifestations, which include adiposity of the face and trunk, but not the limbs, weakness, fatigue, abdominal striae, facial hair, high blood pressure and raised blood sugar. Although I spent two years in an army neurosurgical unit, I never saw a case. More frequently, the same clinical picture is produced by a benign adenoma of the suprarenal cortex, but today by far the commonest cause of this condition is seen in the patient receiving high dosage of cortisone, when we talk about the person exhibiting 'Cushingoid features'. Most readers of this journal will have seen such cases.

  4. Histopathological study of feline eosinophilic dermatoses.

    Science.gov (United States)

    Fondati, A; Fondevila, D; Ferrer, L

    2001-12-01

    A retrospective study was conducted on skin specimens from 24 cats with eosinophilic granuloma complex. The specimens were stained with haematoxylin and eosin and Gallego's trichrome stain. In all specimens, flame figures and/or large foci of so-called "collagen degeneration" were detected and histopathological features were not predictive of the clinical picture. Use of the term eosinophilic dermatosis was advocated in diagnostic dermatopathology. On trichrome-stained sections, normally stained collagen fibres were identified in the middle of both flame figures and large foci of "collagen degeneration" and the debris surrounding collagen bundles showed the same tinctorial properties as eosinophil granules. Eosinophil degranulation around collagen bundles seemed to represent the major pathogenetic event in these lesions, analogous with human flame figures. The term flame figures might therefore be more accurately used to designate those foci of eosinophilic to partly basophilic debris commonly referred to as "collagen degeneration".

  5. Design and synthesis of a vialinin A analog with a potent inhibitory activity of TNF-α production and its transformation into a couple of bioprobes.

    Science.gov (United States)

    Ye, Yue Qi; Onose, Jun-ichi; Abe, Naoki; Koshino, Hiroyuki; Takahashi, Shunya

    2012-04-01

    Vialinin A (1) is an extremely potent inhibitor against tumor necrosis factor (TNF)-α production in rat basophilic leukemia (RBL-2H3) cells. This Letter describes the design and synthesis of its advanced analog, 5',6'-dimethyl-1,1':4'1″-terphenyl-2',3',4,4″-tetraol (2) with a comparable inhibitory activity (IC(50)=0.02 nM) to that of 1. The synthesis involved double Suzuki-Miyaura coupling as a key step, and required only five steps from commercially available 3,4-dimethylphenol. For identification of the target molecule, fluorescent and biotinylated derivatives of 2 were prepared through a 'click' coupling process.

  6. Inclusion body hepatitis (IBH outbreak associated with fowl adenovirus type 8b in broilers

    Directory of Open Access Journals (Sweden)

    Zadravec M.

    2013-01-01

    Full Text Available The causative agent of inclusion body hepatitis (IBH was identified as fowl adenovirus (FAdV type 8b, a member of the Fowl adenovirus E species, based on PCR results of adenoviral polymerase and the hexon gene in an outbreak of acute mortality that affected a broiler flock of 12,000 animals. In two waves of elevated mortality rate, a total of 264 chickens were found dead. Affected birds showed ruffled feathers, depression, watery droppings and limping. The most common pathological lesions seen on necropsy were pale, swollen and friable livers. On histological examination, acute hepatitis characterized by necrosis of hepatocytes, with large basophilic intranuclear inclusion bodies, were observed. In addition, infectious bursal disease virus and infectious bronchitis virus were detected in the same flock.

  7. Human, recombinant interleukin-2 induces in vitro histamine release in a dose-dependent manner

    DEFF Research Database (Denmark)

    Nielsen, Hans Jørgen; Petersen, L J; Skov, P S

    1995-01-01

    We previously observed that human, recombinant interleukin-2 in a pharmacologic dose (200 u/ml) induced histamine release from monocyte-depleted peripheral blood mononuclear cells in vitro. Therefore, we studied the role of various pharmacologic doses of rIL-2 on in vitro histamine release....... Peripheral blood mononuclear cells (5 x 10(6) cells/ml), which also contain basophils, from 13 patients scheduled for elective colorectal cancer surgery and 10 age and sex matched healthy volunteers were stimulated with rIL-2 in concentrations of 0, 50, 100, 200, 450, 900, 1,800 and 3,600 u/ml, respectively......, for 1, 24 and 48 hours under standard conditions. Histamine was analysed in supernatants using the glass fiber method. Simultaneously, total cell-bound histamine was analysed in lysate from 5 x 10(6) mononuclear cells from all patients and volunteers, thus allowing determination of percent histamine...

  8. Oral leiomyomatous hamartoma of the median maxillary gingiva: a case report and review of the literature.

    Science.gov (United States)

    ALQahtani, Dalal; Qannam, Ahmed

    2013-08-01

    Oral leiomyomatous hamartoma (OLH) is a rare lesion seen in the oral cavity. It mainly presents on the median maxilla and tongue. In the literature in English, there are only 10 reported cases of OLH of the median maxilla. Most of the cases were found in patients of Japanese and Latin American origin. We report a case of OLH in an 18-month-old boy of Middle Eastern ancestry. The lesion presented as a pedunculated, light pink, soft swelling that was located on the labial gingiva of tooth number 21. Microscopically, it showed proliferative smooth-muscle fascicles dispersed in loose fibrous stroma and multiple small vessels. The lesional cells looked mature and elongated and were deeply eosinophilic spindle cells with basophilic, central "cigar-shaped" nuclei. The diagnosis of OLH was supported by positive immunohistochemical reactivity of smooth-muscle actin and desmin. To our knowledge, this is the first reported case of OLH in a Middle Eastern patient.

  9. Fifty years later: Emerging functions of IgE antibodies in host defense, immune regulation, and allergic diseases.

    Science.gov (United States)

    Oettgen, Hans C

    2016-06-01

    Fifty years ago, after a long search, IgE emerged as the circulating factor responsible for triggering allergic reactions. Its extremely low concentration in plasma created significant hurdles for scientists working to reveal its identity. We now know that IgE levels are invariably increased in patients affected by atopic conditions and that IgE provides the critical link between the antigen recognition role of the adaptive immune system and the effector functions of mast cells and basophils at mucosal and cutaneous sites of environmental exposure. This review discusses the established mechanisms of action of IgE in pathologic immediate hypersensitivity, as well as its multifaceted roles in protective immunity, control of mast cell homeostasis, and its more recently revealed immunomodulatory functions.

  10. Histopathological changes in the head kidney induced by cadmium in a neotropical fish Colossoma macropomum.

    Science.gov (United States)

    Salazar-Lugo, R; Vargas, A; Rojas, L; Lemus, M

    2013-01-01

    We evaluated the effect of cadmium (Cd) on the structure and function of the head kidney in the freshwater fish Colossoma macropomum (C. macropomum). Juveniles were exposed to 0.1 mg/L CdCl2 for 31 days. Blood samples were examined using hematological tests and head kidney histology was determined by light microscopy. The concentration of Cd in the head and trunk kidneys was measured using an atomic absorption spectrophotometer. Cd produced histopathological changes in the head kidney, the most evident of these being: the thickening of the vein wall, an increase in the number of basophils/mast cells close to blood vessels and a severe depletion of hematopoietic precursors especially the granulopoietic series. In the blood, a decrease in the total leucocytes and hemoglobin concentration was observed. Cd-exposed fish showed higher Cd concentrations in the trunk kidney than the head kidney. In conclusion, exposure to Cd affected precursor hematopoietic cells in C. macropomum.

  11. Polyphenol Interactions Mitigate the Immunogenicity and Allergenicity of Gliadins.

    Science.gov (United States)

    Pérot, Maxime; Lupi, Roberta; Guyot, Sylvain; Delayre-Orthez, Carine; Gadonna-Widehem, Pascale; Thébaudin, Jean-Yves; Bodinier, Marie; Larré, Colette

    2017-03-01

    Wheat allergy is an IgE-mediated disorder. Polyphenols, which are known to interact with certain proteins, could be used to reduce allergic reactions. This study screened several polyphenol sources for their ability to interact with gliadins, mask epitopes, and affect basophil degranulation. Polyphenol extracts from artichoke leaves, cranberries, apples, and green tea leaves were examined. Of these extracts, the first three formed insoluble complexes with gliadins. Only the cranberry and apple extracts masked epitopes in dot blot assays using anti-gliadin IgG and IgE antibodies from patients with wheat allergies. The cranberry and artichoke extracts limited cellular degranulation by reducing mouse anti-gliadin IgE recognition. In conclusion, the cranberry extract is the most effective polyphenol source at reducing the immunogenicity and allergenicity of wheat gliadins.

  12. Haematological studies of Gaolao Heifers

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    P.M.Kapale,D.G.Jagtap

    2008-02-01

    Full Text Available Hematological study of 15 healthy Gaolao heifers has been carried out. The mean ±S.E. values of total erythrocyte (10.3 ±0.34 million/cu mm, Hb (11.92 ±0.33 gm%, PCV (34.07±1.11%, MCV (33.14±0.75cuμ, MCH (11.63±0.32 μμgm, MCHC (35.07 ±0.70%,TLC (9.51±0.92 thousand/cmm, Lymphocytes (64.93 ±1.74%,Neutrophil (28.93 ±1.8%, Monocyte (2.2 ±0.34%, Basophil (0.2±0.11 and Eosinophil (3.7 ± 0.44% have been reported. [Veterinary World 2008; 1(1.000: 16-16

  13. Haematological Studies of Gaolao Calves

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    P. M. Kapale

    2008-02-01

    Full Text Available The hematological study in fifteen healthy Gaolao calves ranging from birth to 12 months of age indicate the mean ± S.E. value of TEC (11.00± 0.20 million/cu.mm, Hb (14.27±0.43 gm%, PCV (35.00±0.79%, MCV (31.70±0.92 cu.μ, MCH (13.04±0.84 μ. μgm, MCHC (41.16±1.76%, TLC (07.49±0.39 thousand/cu.mm, Lymphocytes (71.87 ±1.45%, Neutrophil (23.27±1.27%, Monocyte (1.3 ±0.25%, Basophil (0.27±0.12% and Esonophil (3.4 ±0.49%. [Veterinary World 2008; 1(1.000: 17-17

  14. Several distinct properties of the IgE repertoire determine effector cell degranulation in response to allergen challenge

    DEFF Research Database (Denmark)

    Christensen, Lars Harder; Holm, Jens-Christian; Lund, Gitte;

    2008-01-01

    BACKGROUND: On cross-linking of receptor-bound IgE antibodies by allergens, effector cells (basophils and mast cells) involved in type I allergic reactions degranulate and release the potent chemical mediators stored inside their granules. Total and allergen-specific IgE concentrations, Ig......E affinity for allergen, and IgE clonality are all distinct properties of allergic patients' IgE repertoires. However, the inability to isolate individual IgE antibodies from allergic patients' sera presents a major barrier to understanding the importance of patient-specific IgE repertoires...... for the manifestation and severity of allergic symptoms. OBJECTIVE: We sought to investigate how individual properties of an IgE repertoire affect effector cell degranulation. METHODS: A panel of recombinant IgE (rIgE) antibodies specific for the major house dust mite allergen Der p 2 was developed and characterized...

  15. Aberrant expression of regulatory cytokine IL-35 and pattern recognition receptor NOD2 in patients with allergic asthma.

    Science.gov (United States)

    Wong, Chun Kwok; Leung, Ting Fan; Chu, Ida Miu Ting; Dong, Jie; Lam, Yvonne Yi On; Lam, Christopher Wai Kei

    2015-02-01

    We investigated the plasma concentration of the novel regulatory cytokine IL-35 and intracytosolic pattern recognition receptors nucleotide-binding oligomerization domain (NOD)-like receptors in granulocytes and explored their potential implication in disease severity monitoring of allergic asthma. The expression of circulating IL-35 and other pro-inflammatory mediators in asthmatic patients or control subjects were evaluated using enzyme-linked immunosorbent assay (ELISA). The intracellular expressions of NOD1 and NOD2 in CCR3+ granulocytes were assessed using flow cytometry. Plasma concentrations of IL-35, IL-17A, basophil activation marker basogranulin, and eosinophilic airway inflammation biomarker periostin were significantly elevated in allergic asthmatic patients compared to non-atopic control subjects (all probability (p) IL-35 concentration in asthmatic patients (all p IL-35 and periostin with disease severity score in asthmatic patients (both p IL-35 (p IL-35 may serve as a potential surrogate biomarker for disease severity of allergic asthma.

  16. Morphologic, cytometric and functional characterization of the common octopus (Octopus vulgaris) hemocytes.

    Science.gov (United States)

    Castellanos-Martínez, S; Prado-Alvarez, M; Lobo-da-Cunha, A; Azevedo, C; Gestal, C

    2014-05-01

    The hemocytes of Octopus vulgaris were morphologically and functionally characterized. Light and electron microscopy (TEM and SEM), and flow cytometry analyses revealed the existence of two hemocyte populations. Large granulocytes showed U-shaped nucleus, a mean of 11.6 μm±1.2 in diameter with basophilic granules, polysaccharide and lysosomic deposits in the cytoplasm. Small granulocytes measured a mean of 8.1 μm±0.7 in diameter, and have a round nucleus occupying almost the entire cell and few or not granules in the cytoplasm. Flow cytometry analysis showed that large granulocytes are the principal cells that develop phagocytosis of latex beads (rising up to 56%) and ROS after zymosan stimulation. Zymosan induced the highest production of both ROS and NO. This study is the first tread towards understanding the O. vulgaris immune system by applying new tools to provide a most comprehensive morpho-functional study of their hemocytes.

  17. Japanese Guideline for Food Allergy 2014

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    Atsuo Urisu

    2014-01-01

    The therapy for food allergies includes treatment of and prophylactic measures against hypersensitivity such as anaphylaxis. A fundamental prophylactic measure is the elimination diet. However, elimination diets should be used only if necessary because of the patient-related burden. For this purpose, it is very important that causative foods be accurately identified. There are a number of means available to identify causative foods, including the history taking, a skin prick test, detection of antigen-specific IgE antibodies in the blood, the basophil histamine release test, the elimination diet test, and the oral challenge test, etc. Of these, the oral challenge test is the most reliable. However, it should be conducted under the supervision of experienced physicians because it may cause adverse reactions, such as anaphylaxis.

  18. Japanese Guideline for Food Allergy

    Directory of Open Access Journals (Sweden)

    Atsuo Urisu

    2011-01-01

    Therapy for food allergy includes treatments of and prophylactic measures against hypersensitivity like anaphylaxis. A fundamental prophylactic measure is the elimination diet. However, elimination diets should be conducted only if they are inevitable because they places a burden on patients. For this purpose, it is highly important that causative foods are accurately identified. Many means to determine the causative foods are available, including history taking, skin prick test, antigen specific IgE antibodies in blood, basophil histamine release test, elimination diet test, oral food challenge test, etc. Of these, the oral food challenge test is the most reliable. However, it should be conducted under the supervision of experienced physicians because it may cause adverse reactions such as anaphylaxis.

  19. Avian polyomavirus infection of a fledgling budgerigar (Melopsittacus undulatus) and differential diagnoses of viral inclusions in psittacine birds--case report and mini-review.

    Science.gov (United States)

    Herder, Vanessa; König, Anett; Seehusen, Frauke; Wohlsein, Peter

    2011-01-01

    A two-week-old budgerigar (Melopsittacus undulatus) of an outdoor aviary died suddenly and was submitted for determination the cause of illness and death. Macroscopically, the sparsely feathered animal was in a poor body condition. Histopathological examination revealed in various mesenchymal and epithelial tissues, numerous up to 15 microm in diameter large intranuclear, amphophilic to basophilic inclusion bodies with a clearing of the centre. Additionally, a feather dysplasia and retention hyperkeratosis of feather follicles was found. Ultrastructurally, viral particles of approximately 35 nm in diameter were detected in the feather follicle epithelium. A PCR for Avian Polyomavirus on fresh skin samples was negative whereas on formalin-fixed kidney samples with a high amount of viral inclusion bodies yielded a positive result. In addition, viral inclusion body diseases, like Avian Poxvirus, Psittacine Beak and Feather disease virus, Avian Adenovirus, Psittacine Herpesvirus and papillomavirus of psittacines are summarized and compared in the present article.

  20. Haematological characteristics of freshwater-reared and wild mullet, Mugil platanus Günther (Osteichthyes, Mugilidae

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    Maria José T Ranzani-Paiva

    1996-01-01

    Full Text Available Mullets, Mugilplalanus Günther, 1880. were obtained from the estua-rine area of CananÉia (São Paulo, Brazil and from freshwater tearing tanks. For each fish the body weight, body length, gonad weight and gonad maturation stage were recorded and the allometric index (K was calculated. Blood was collected for hematocrit (Ht, hemoglobin level (lib, total cell count (Er, and the determination of hematological indices (MCV. MCH, MCHC, as well as the differential leukocyte count (lymphocytes, monocytes, neutrophils, basophils and '"other cells". The averages of the hematological analyses, hematological indeces, condition factor and leukocytes percentage of the specimens acclimated to fresh water were significantly higher than those of the specimens from the estuary.

  1. ROLE OF D2 DOPAMINE RECEPTOR ON MODULATION OF THE LEUKOCYTE FORMULA IN RESTRAINT STRESSED RATS

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    Lucian Hritcu

    2006-08-01

    Full Text Available Dopamine is a monoamine neurotransmitter of both central and peripheral nervous system. Its role in the neural-immune communication has been discussed in the present study. Results reveal that in vivo blockade of D2 dopamine receptor by means of sulpiride, a selective antagonist for D2 dopamine receptor produce changes in functional activities of the immune effector cells. Adults rats pretreated once with LPS (a bacterial product (25µg/250µl, i.p., produce an immune response, were subjected to i.p. injection with sulpiride (4 mg/kg b.w., i.p., a selective antagonist for D2 dopamine receptors, after 3 days postimmunization. After 18 days later, we assessed the total leukocyte number, neutrophils, eosinophils and basophils number. In summary, we provide that D2 dopamine receptor blockade suppress or enhance the immune effector cells number in restraint stress.

  2. Of ITIMs, ITAMs, and ITAMis: revisiting immunoglobulin Fc receptor signaling.

    Science.gov (United States)

    Getahun, Andrew; Cambier, John C

    2015-11-01

    Receptors for immunoglobulin Fc regions play multiple critical roles in the immune system, mediating functions as diverse as phagocytosis, triggering degranulation of basophils and mast cells, promoting immunoglobulin class switching, and preventing excessive activation. Transmembrane signaling associated with these functions is mediated primarily by two amino acid sequence motifs, ITAMs (immunoreceptor tyrosine-based activation motifs) and ITIMs (immunoreceptor tyrosine-based inhibition motifs) that act as the receptors' interface with activating and inhibitory signaling pathways, respectively. While ITAMs mobilize activating tyrosine kinases and their consorts, ITIMs mobilize opposing tyrosine and inositol-lipid phosphatases. In this review, we will discuss our current understanding of signaling by these receptors/motifs and their sometimes blurred lines of function.

  3. 水疱型毛母质瘤1例%Pilomatricoma with Bullous Appearance:A Case Report

    Institute of Scientific and Technical Information of China (English)

    孔玉龙; 王克玉

    2014-01-01

    患者,男,12岁。右侧颈部及右上臂囊性肿物2年。皮损组织病理示:表皮正常,真皮内见镜影细胞及嗜碱性粒细胞浸润,可见多处钙化区,其周围被纤维组织包绕。诊断:水疱型毛母质瘤。%A 12-year-old male patient presented with purplish red bullous on right neck and right upper arm. The histopathology of the skin lession showed a normal epidermis. Shadow cells and basophilic cell infil-tration with calcifying areas, surrounded by fibrous tissue. He was diagnosed as pilomatricoma with bullous appearance.

  4. Clinical and diagnostic study of trypanosomiasis of cattle in mosul, Iraq

    Directory of Open Access Journals (Sweden)

    M. S. H. Rhaymah; B. A. AL-Badrani

    2012-02-01

    Full Text Available This study reported the occurrence of Trypanosoma vivax in cattle in Mosul, Iraq and its effect on their haemotology. The main clinical sings were anemia, lethargy, weakness, general weight loss and, loss of appetite. Blood samples from infected cattle were collected which showed decrease of hemoglobin concentration, packed cell volume and total erythrocytes count with leucopenia especially neutropenia and esonopenia. Blood smears showed short form of trypanosome in acute cases and long forms in chronic cases. Morphological changes of erythrocytes showed the presence of anisocytosis, poikelocytosis, target cells, macrocytes, punctuate basophile and sometimes monocytes. Anemia was basically normocytic normochromic. This study provides the baseline data for further studies on this parasite.

  5. Anatomia e histologia do conduto genital feminino de Pomacea canaliculata (Lamarck, 1822 (Mollusca, Gastropoda, Pilidae

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    Eliana de Fátima Marques de Mesquita

    1990-01-01

    Full Text Available In this paper the authors give an anatomical and histological analysis of the genital duct in mature and immature females of Pomacea canaliculata (Lamarck, 1822. Anatomically, the vagina has an equal dimension in its extension, varying in form and volume, according to the maturation period. In the immaturity, it has a smaller diameter and volume. In the maturity, the vagina increases in volume, having the aspect of a tumescent organ, and in certain specimens shows an albuminous pink thread in its lumen. Histologically, the calcigenic activity of the gland is evidentiated by the presence of an amorphous and basophilic mass, without nuclear material. This material has a fragmented aspect in the vagina lumen that reacts positively in van Kossa's coloured preparations for calcareous salts.

  6. Metastatic Calcinosis Cutis: A Case in a Child with Acute Pre-B Cell Lymphoblastic Leukemia

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    Juan Pablo Castanedo-Cázares

    2015-01-01

    Full Text Available Hypercalcemia in children with malignancy is an uncommon condition. It has been described in leukemia patients with impaired renal excretion of calcium or osteolytic lesions. Metastatic calcinosis cutis (MCC may develop if hypercalcemia persists. We report the case of a 5-year-old girl with an atypical dermatosis and unspecific gastrointestinal symptoms. Considered clinical diagnoses were xanthomas, histiocytosis, molluscum contagiosum, and nongenital warts. Cutaneous histological analysis showed amorphous basophilic deposits in the dermis suggestive of calcium deposits. Laboratory tests confirmed serum hypercalcemia. Extensive investigations such as bone marrow biopsy established the diagnosis of an acute pre-B cell lymphoblastic leukemia. Hypercalcemia in hematopoietic malignancies is unusual, especially as initial manifestation of the disease. Careful review of the literature fails to reveal previous reports of these peculiar cutaneous lesions of MCC in children with leukemia.

  7. Morphologic and cytochemical characteristics of green turtle (Chelonia mydas) blood cells

    Science.gov (United States)

    Work, T.M.; Raskin, R.E.; Balazs, G.H.; Whittaker, S.D.

    1998-01-01

    Objective - To identify and characterize blood cells from free-ranging Hawaiian green turtles, Chelonia mydas. Sample Population - 26 green turtles from Puako on the island of Hawaii and Kaneohe Bay on the island of Oahu. Procedure - Blood was examined, using light and electron microscopy and cytochemical stains that included benzidine peroxidase, chloroacetate esterase, alpha naphthyl butyrate esterase, acid phosphatase, Sudan black B, periodic acid-Schiff, and toluidine blue. Results - 6 types of WBC were identified: lymphocytes, monocytes, thrombocytes, heterophils, basophils, and eosinophils (small and large). Morphologic characteristics of mononuclear cells and most granulocytes were similar to those of cells from other reptiles except that green turtles have both large and small eosinophils. Conclusions - Our classification of green turtle blood cells clarifies imporoper nomenclature reported previously and provides a reference for future hematologic studies in this species.

  8. A 13-Month-Old With Xanthogranulomatous Pyelonephritis With Features of Renal Malakoplakia

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    Tova Appleson DO

    2016-02-01

    Full Text Available Xanthogranulomatous pyelonephritis is an uncommon chronic inflammatory renal disorder caused by chronic infection with gram-negative bacteria leading to destruction of the renal parenchyma and replacement with foamy lipid-laden macrophages. Renal malakoplakia is another rare form of chronic inflammatory granulomatous disease in the kidney associated with infection usually occurring in adults with immunocompromised status or debilitating disease. It is hallmarked by the finding of foamy histiocytes with distinctive basophilic inclusions (Michaelis-Gutmann bodies. We present a case of a 13-month-old male with history of congenital hydronephrosis who presented with clinical and radiologic findings suggestive of xanthogranulomatous pyelonephritis. However, further pathologic studies revealed the presence of Michaelis-Gutmann bodies, which are pathognomonic for renal malakoplakia. With this case we hope to bring further evidence to support that these two conditions are not mutually exclusive but rather represent two pathologic processes on the same disease spectrum.

  9. Monoterpene derivatives with anti-allergic activity from red peony root, the root of Paeonia lactiflora.

    Science.gov (United States)

    Shi, Yan-Hong; Zhu, Shu; Ge, Yue-Wei; He, Yu-Min; Kazuma, Kohei; Wang, Zhengtao; Yoshimatsu, Kayo; Komatsu, Katsuko

    2016-01-01

    The methanolic extract and its subfractions from red peony root, the dried roots of Paeonia lactiflora Pallas showed potent antiallergic effects, as inhibition of immunoglobulin E (IgE)-mediated degranulation in rat basophil leukemia (RBL)-2H3 cells. Bioassay-guided fractionation led to the isolation of 16 monoterpene derivatives, including 3 new compounds, paeoniflorol (1), 4'-hydroxypaeoniflorigenone (2) and 4-epi-albiflorin (3), together with 13 known ones (4-16). The chemical structures of the new compounds were elucidated on the basis of spectroscopic and chemical evidences. Among the isolated monoterpene derivatives, nine compounds showed potent anti-allergic effects and compound 1 was the most effective. A primary structure-activity relationship of monoterpene derivatives was discussed.

  10. Omalizumab is efficacious for management of recalcitrant, antihistamine-resistant chronic urticaria.

    Science.gov (United States)

    Lang, D M

    2015-06-01

    Chronic urticaria continues to be a challenging condition for both patients and physicians. Despite improved understanding of chronic urticaria, many patients continue to experience ongoing symptoms and impaired quality of life. Omalizumab is a recombinant humanized monoclonal antibody that binds to the domain at which IgE binds to the high-affinity IgE receptor on mast cells and basophils. The efficacy of omalizumab for antihistamine-resistant chronic urticaria has been demonstrated in several randomized controlled trials as well as observational studies. Omalizumab is generally well tolerated, and is associated with less potential for harm compared with other therapeutic alternatives (e.g., calcineurin inhibitors) for recalcitrant chronic urticaria. Omalizumab has become the best-studied agent for treatment of antihistamine-resistant chronic urticaria, and the agent for which the data in support of its efficacy is most methodologically sound. Omalizumab is an effective therapeutic option for patients with recalcitrant chronic urticaria.

  11. Effect of simulated gastro-duodenal digestion on the allergenic reactivity of beta-lactoglobulin

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    Bossios Apostolos

    2011-08-01

    Full Text Available Abstract Background Cow's milk (CM allergy affects about 2% of infants. The allergenicity of dietary proteins, including those from CM, has been related to their digestibility although the generality of the link and its causality remains to be demonstrated. In this study we use an in vitro digestion system, to investigate the digestibility of β-lactoglobulin (blg during gastrointestinal transit and to assess the impact of this process on blg allergenic reactivity in CM allergic children. Methods Blg digesta were prepared using an in vitro digestion protocol simulating either gastric digestion alone or followed by duodenal digestion with or without phosphatidylcholine (PC. Biochemical analysis of blg digesta was performed by SDS-PAGE and their concentration was measured by a sandwich ELISA. Assessment of their allergenic reactivity was done in vitro by EAST inhibition, specific basophil activation (basotest and lymphocyte proliferation (PCNA-flow cytometry assays using sera and cells from patients allergic to blg and in vivo by skin prick testing (SPT of these patients. Results Blg was only broken down to smaller peptides after gastro-duodenal digestion although a sizeable amount of intact protein still remained. Digestion did not modify the IgE binding capacity of blg except for gastro-duodenal digestion performed in the absence of PC. These results are consistent with the quantity of intact blg remaining in the digesta. Overall both gastric and gastroduodenal digestion enhanced activation of sensitized basophils and proliferation of sensitized lymphocytes by blg. However, there was a tendency towards reduction in mean diameter of SPT following digestion, the PC alone during phase 1 digestion causing a significant increase in mean diameter. Conclusions Digestion did not reduce the allergenic reactivity of blg to a clinically insignificant extent, PC inhibiting digestion and thereby protecting blg allergenic reactivity. SPT reactivity was

  12. Functional role of human NK cell receptor 2B4 (CD244) isoforms.

    Science.gov (United States)

    Mathew, Stephen O; Rao, Krithi K; Kim, Jong R; Bambard, Nowland D; Mathew, Porunelloor A

    2009-06-01

    2B4 (CD244), a member of the signaling lymphocyte-activation molecule (SLAM/CD150), is expressed on all NK cells, a subpopulation of T cells, monocytes and basophils. Human NK cells express two isoforms of 2B4, h2B4-A and h2B4-B that differ in a small portion of the extracellular domain. In the present investigation, we have studied the functions of h2B4-A and h2B4-B. Our study demonstrated that these two isoforms differ in their binding affinity for CD48, which results in differential cytotoxic activity as well as intracellular calcium release by NK cells upon target cell recognition. Analysis of the predicted 3-D structure of the two isoforms showed conformational differences that could account for their differences in binding affinity to CD48. h2B4-A was able to mediate natural cytotoxicity against CD48-expressing K562 target cells and induce intracellular calcium release, whereas h2B4-B showed no effects. NK-92MI, U937, THP-1, KU812, primary monocytes, basophils and NK cells showed expression of both h2B4-A and h2B4-B whereas YT and IL-2-activated NK cells did not show any h2B4-B expression. Stimulation of NK cells through 2B4 resulted in decreased mRNA levels of both h2B4-A and h2B4-B indicating that down-regulation of 2B4 isoforms may be an important factor in controlling NK cell activation during immune responses.

  13. Genetic engineering of the major timothy grass pollen allergen, Phl p 6, to reduce allergenic activity and preserve immunogenicity.

    Science.gov (United States)

    Vrtala, Susanne; Focke, Margarete; Kopec, Jolanta; Verdino, Petra; Hartl, Arnulf; Sperr, Wolfgang R; Fedorov, Alexander A; Ball, Tanja; Almo, Steve; Valent, Peter; Thalhamer, Josef; Keller, Walter; Valenta, Rudolf

    2007-08-01

    On the basis of IgE epitope mapping data, we have produced three allergen fragments comprising aa 1-33, 1-57, and 31-110 of the major timothy grass pollen allergen Phl p 6 aa 1-110 by expression in Escherichia coli and chemical synthesis. Circular dichroism analysis showed that the purified fragments lack the typical alpha-helical fold of the complete allergen. Superposition of the sequences of the fragments onto the three-dimensional allergen structure indicated that the removal of only one of the four helices had led to the destabilization of the alpha helical structure of Phl p 6. The lack of structural fold was accompanied by a strong reduction of IgE reactivity and allergenic activity of the three fragments as determined by basophil histamine release in allergic patients. Each of the three Phl p 6 fragments adsorbed to CFA induced Phl p 6-specific IgG Abs in rabbits. However, immunization of mice with fragments adsorbed to an adjuvant allowed for human use (AluGel-S) showed that only the Phl p 6 aa 31-110 induced Phl p 6-specific IgG Abs. Anti-Phl p 6 IgG Abs induced by vaccination with Phl p 6 aa 31-110 inhibited patients' IgE reactivity to the wild-type allergen as well as Phl p 6-induced basophil degranulation. Our results are of importance for the design of hypoallergenic allergy vaccines. They show that it has to be demonstrated that the hypoallergenic derivative induces a robust IgG response in a formulation that can be used in allergic patients.

  14. Heme oxygenase-1 deletion affects stress erythropoiesis.

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    Yu-An Cao

    Full Text Available BACKGROUND: Homeostatic erythropoiesis leads to the formation of mature red blood cells under non-stress conditions, and the production of new erythrocytes occurs as the need arises. In response to environmental stimuli, such as bone marrow transplantation, myelosuppression, or anemia, erythroid progenitors proliferate rapidly in a process referred to as stress erythropoiesis. We have previously demonstrated that heme oxygenase-1 (HO-1 deficiency leads to disrupted stress hematopoiesis. Here, we describe the specific effects of HO-1 deficiency on stress erythropoiesis. METHODOLOGY/PRINCIPAL FINDINGS: We used a transplant model to induce stress conditions. In irradiated recipients that received hmox(+/- or hmox(+/+ bone marrow cells, we evaluated (i the erythrocyte parameters in the peripheral blood; (ii the staining intensity of CD71-, Ter119-, and CD49d-specific surface markers during erythroblast differentiation; (iii the patterns of histological iron staining; and (iv the number of Mac-1(+-cells expressing TNF-α. In the spleens of mice that received hmox(+/- cells, we show (i decreases in the proerythroblast, basophilic, and polychromatophilic erythroblast populations; (ii increases in the insoluble iron levels and decreases in the soluble iron levels; (iii increased numbers of Mac-1(+-cells expressing TNF-α; and (iv decreased levels of CD49d expression in the basophilic and polychromatophilic erythroblast populations. CONCLUSIONS/SIGNIFICANCE: As reflected by effects on secreted and cell surface proteins, HO-1 deletion likely affects stress erythropoiesis through the retention of erythroblasts in the erythroblastic islands of the spleen. Thus, HO-1 may serve as a therapeutic target for controlling erythropoiesis, and the dysregulation of HO-1 may be a predisposing condition for hematologic diseases.

  15. Interaction between endothelial cells and the secreted cytokine drives the fate of an IL4- or an IL5-transduced tumour.

    Science.gov (United States)

    Di Carlo, E; Modesti, A; Coletti, A; Colombo, M P; Giovarelli, M; Forni, G; Diodoro, M G; Musiani, P

    1998-12-01

    Injection of interleukin-4 (IL4) gene-transduced tumour cells into syngeneic immunocompetent mice resulted in tumour rejection in which a key role for eosinophils was suggested. To evaluate whether IL5 inhibits tumour growth by selectively inducing eosinophil recruitment and activation, a poorly differentiated mammary adenocarcinoma cell line (TSA) was transfected with the IL5 gene and the cells secreting IL5 (TSA-IL5) were injected subcutaneously (s.c.) in syngeneic mice. The oncogenicity of TSA-IL5 was compared with that exhibited by TSA cells transfected with the IL4 gene (TSA-IL4) and with the neomycin resistance gene only (TSA-neo). At progressive times after subcutaneous challenge, tumour growth areas were studied histologically, ultrastructurally, and immunohistochemically to identify the reactive cells, visualize tumour vessels, and detect the cytokines and chemokines involved in the anti-tumour reaction. Both the morphological and the functional data showed that TSA-IL5, despite the large eosinophil infiltrate, grew progressively like TSA-neo, suggesting that eosinophils per se do not play a crucial role in TSA tumour rejection. Furthermore, our data indicate that the rejection of TSA-IL4 depends on the IL4-induced expression of VCAM-1 and MCP-1 by endothelial cells. MCP-1 together with VCAM-1 results in recruitment and activation of basophils, mast cells, and macrophages, and hence a pro-inflammatory cytokine cascade that initially favours the influx and activation of neutrophils and finally tumour rejection. In this context, the rejection of TSA-IL4 seems to involve a variety of reactive cells and rests on a continuous cross-talk between basophils, mast cells, macrophages, CD8-positive lymphocytes, and granulocyte subsets, mostly neutrophils.

  16. Antigenic proteins involved in occupational rhinitis and asthma caused by obeche wood (Triplochiton scleroxylon.

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    Ana Aranda

    Full Text Available BACKGROUND: Obeche wood dust is a known cause of occupational asthma where an IgE-mediated mechanism has been demonstrated. OBJECTIVE: To characterize the allergenic profile of obeche wood dust and evaluate the reactivity of the proteins by in vitro, ex vivo and in vivo assays in carpenters with confirmed rhinitis and/or asthma MATERIALS AND METHODS: An in-house obeche extract was obtained, and two IgE binding bands were purified (24 and 12 kDa and sequenced by N-terminal identity. Specific IgE and IgG, basophil activation tests and skin prick tests (SPTs were performed with whole extract and purified proteins. CCD binding was analyzed by ELISA inhibition studies. RESULTS: Sixty-two subjects participated: 12 with confirmed occupational asthma/rhinitis (ORA+, 40 asymptomatic exposed (ORA-, and 10 controls. Of the confirmed subjects, 83% had a positive SPT to obeche. There was a 100% recognition by ELISA in symptomatic subjects vs. 30% and 10% in asymptomatic exposed subjects and controls respectively (p<0.05. Two new proteins were purified, a 24 kDa protein identified as a putative thaumatin-like protein and a 12 kDa gamma-expansin. Both showed allergenic activity in vitro, with the putative thaumatin being the most active, with 92% recognition by ELISA and 100% by basophil activation test in ORA+ subjects. Cross-reactivity due to CCD was ruled out in 82% of cases. CONCLUSIONS: Two proteins of obeche wood were identified and were recognized by a high percentage of symptomatic subjects and by a small proportion of asymptomatic exposed subjects. Further studies are required to evaluate cross reactivity with other plant allergens.

  17. Time-course changes in nasal airway resistance after house dust antigen challenge: With special reference to late phase response

    Directory of Open Access Journals (Sweden)

    Kimihiro Okubo

    1998-01-01

    Full Text Available The definition of late-phase response (LPR associated with nasal allergy is not as clear as that associated with asthma. Furthermore, LPR and immediate-phase response often act in concert to produce confounding symptoms due to repeated attacks over a short period of time. We examined the nasal airway resistance (NAR, allergic symptoms, eosinophil cationic protein and histamine concentration in nasal surface tissue 30 min before, and 10 min, 1, 3, 5, 7 and 9 h after a house dust (HD nasal challenge test was performed twice on 10 patients with HD perennial nasal allergy. Nasal airway resistance readings after the HD nasal challenge test were classified into three types based on changes in NAR: type I (short-lasting, five cases; type II (prolonged, eight cases; and type III (biphasic, six cases. Delayed increases in NAR were not observed in type I patients with weak responses to nasal challenge. In type II patients, baseline NAR was elevated and numerous basophilic metachromatic cells (BC were present in the epithelial layer. In type III patients, baseline NAR was not as elevated as it was in type II patients, but marked responses to challenge were noted. In all three types of changes in NAR, eosinophil leukocyte increased at late phase. Moreover, there was no correlation of change in NAR with an increase in the number of eosinophil leukocyte and BC at the epithelial layer in all three types, which suggests that further study is required to confirm the exact role of eosinophils and basophilic cells in LPR.

  18. Pichia pastoris is superior to E. coli for the production of recombinant allergenic non-specific lipid-transfer proteins.

    Science.gov (United States)

    Pokoj, Sven; Lauer, Iris; Fötisch, Kay; Himly, Martin; Mari, Adriano; Enrique, Ernesto; Miguel-Moncin, Maria Del Mar San; Lidholm, Jonas; Vieths, Stefan; Scheurer, Stephan

    2010-01-01

    Non-specific lipid-transfer proteins (nsLTP) from food and pollen are clinically important allergens, especially in patients recruited from the Mediterranean area. For the use of recombinant nsLTPs in allergy diagnosis and preclinical allergy studies the preparation of nsLTPs in a properly folded and biologically active form is required. Using hazelnut nsLTP (Cor a 8) as a model allergen, heterologous over-expression in Escherichia coli and Pichia pastoris was compared. Recombinant Cor a 8 derived from E. coli and P. pastoris was purified by IMAC and SEC or ammonium sulphate precipitation followed by IEC and SEC, respectively. The recombinant proteins were characterized with regard to IgE-binding by immunoblotting and ELISA, structure by N-terminal sequencing, CD-spectroscopy and LS and to their biological activity using an in vitro basophil histamine release assay. Purification of hazelnut nsLTP from bacterial lysate under native conditions resulted in a low yield of Cor a 8. In addition, the preparation contained non-IgE-reactive aggregations besides the IgE-reactive monomer. In contrast, the yield of rCor a 8 produced in P. pastoris was approximately 270-fold higher and impurities with oligomers have not been detected. Purified monomeric Cor a 8 from bacteria and yeast showed similar IgE-antibody reactivity and secondary structures, and both were capable of inducing histamine release from basophils. In summary, P. pastoris is superior to E. coli as expression system for the production of large quantities of soluble, properly folded, and biologically active rCor a 8.

  19. Role of IgE in autoimmunity.

    Science.gov (United States)

    Sanjuan, Miguel A; Sagar, Divya; Kolbeck, Roland

    2016-06-01

    There is accumulating evidence to suggest that IgE plays a significant role in autoimmunity. The presence of circulating self-reactive IgE in patients with autoimmune disorders has been long known but, at the same time, largely understudied. However, studies have shown that the increased IgE concentration is not associated with higher prevalence for atopy and allergy in patients with autoimmune diseases, such as systemic lupus erythematosus. IgE-mediated mechanisms are conventionally known to facilitate degranulation of mast cells and basophils and promote TH2 immunity, mechanisms that are not only central to mounting an appropriate defense against parasitic worms, noxious substances, toxins, venoms, and environmental irritants but that also trigger exuberant allergic reactions in patients with allergies. More recently, IgE autoantibodies have been recognized to participate in the self-inflicted damaging immune responses that characterize autoimmunity. Such autoimmune responses include direct damage on tissue-containing autoantigens, activation and migration of basophils to lymph nodes, and, as observed most recently, induction of type 1 interferon responses from plasmacytoid dendritic cells. The importance of IgE as a central pathogenic mechanism in autoimmunity has now been clinically validated by the approval of omalizumab, an anti-IgE mAb, for patients with chronic spontaneous urticaria and for the clinical benefit of patients with bullous pemphigoid. In this review we summarize recent reports describing the prevalence of self-reactive IgE and discuss novel findings that incriminate IgE as central in the pathogenesis of inflammatory autoimmune disorders.

  20. Detection of Chlamydia in the peripheral blood cells of normal donors using in vitro culture, immunofluorescence microscopy and flow cytometry techniques

    Directory of Open Access Journals (Sweden)

    Andrzejewski Chester

    2006-02-01

    Full Text Available Abstract Background Chlamydia trachomatis (Ct and Chlamydia pneumoniae (Cp are medically significant infectious agents associated with various chronic human pathologies. Nevertheless, specific roles in disease progression or initiation are incompletely defined. Both pathogens infect established cell lines in vitro and polymerase chain reaction (PCR has detected Chlamydia DNA in various clinical specimens as well as in normal donor peripheral blood monocytes (PBMC. However, Chlamydia infection of other blood cell types, quantification of Chlamydia infected cells in peripheral blood and transmission of this infection in vitro have not been examined. Methods Cp specific titers were assessed for sera from 459 normal human donor blood (NBD samples. Isolated white blood cells (WBC were assayed by in vitro culture to evaluate infection transmission of blood cell borne chlamydiae. Smears of fresh blood samples (FB were dual immunostained for microscopic identification of Chlamydia-infected cell types and aliquots also assessed using Flow Cytometry (FC. Results ELISA demonstrated that 219 (47.7% of the NBD samples exhibit elevated anti-Cp antibody titers. Imunofluorescence microscopy of smears demonstrated 113 (24.6% of samples contained intracellular Chlamydia and monoclonals to specific CD markers showed that in vivo infection of neutrophil and eosinophil/basophil cells as well as monocytes occurs. In vitro culture established WBCs of 114 (24.8% of the NBD samples harbored infectious chlamydiae, clinically a potentially source of transmission, FC demonstrated both Chlamydia infected and uninfected cells can be readily identified and quantified. Conclusion NBD can harbor infected neutrophils, eosinophil/basophils and monocytes. The chlamydiae are infectious in vitro, and both total, and cell type specific Chlamydia carriage is quantifiable by FC.

  1. Adjuvant effects of aluminium hydroxide-adsorbed allergens and allergoids - differences in vivo and in vitro.

    Science.gov (United States)

    Heydenreich, B; Bellinghausen, I; Lund, L; Henmar, H; Lund, G; Adler Würtzen, P; Saloga, J

    2014-06-01

    Allergen-specific immunotherapy (SIT) is a clinically effective therapy for immunoglobulin (Ig)E-mediated allergic diseases. To reduce the risk of IgE-mediated side effects, chemically modified allergoids have been introduced. Furthermore, adsorbance of allergens to aluminium hydroxide (alum) is widely used to enhance the immune response. The mechanisms behind the adjuvant effect of alum are still not completely understood. In the present study we analysed the effects of alum-adsorbed allergens and allergoids on their immunogenicity in vitro and in vivo and their ability to activate basophils of allergic donors. Human monocyte derived dendritic cells (DC) were incubated with native Phleum pratense or Betula verrucosa allergen extract or formaldehyde- or glutaraldehyde-modified allergoids, adsorbed or unadsorbed to alum. After maturation, DC were co-cultivated with autologous CD4(+) T cells. Allergenicity was tested by leukotriene and histamine release of human basophils. Finally, in-vivo immunogenicity was analysed by IgG production of immunized mice. T cell proliferation as well as interleukin (IL)-4, IL-13, IL-10 and interferon (IFN)-γ production were strongly decreased using glutaraldehyde-modified allergoids, but did not differ between alum-adsorbed allergens or allergoids and the corresponding unadsorbed preparations. Glutaraldehyde modification also led to a decreased leukotriene and histamine release compared to native allergens, being further decreased by adsorption to alum. In vivo, immunogenicity was reduced for allergoids which could be partly restored by adsorption to alum. Our results suggest that adsorption of native allergens or modified allergoids to alum had no consistent adjuvant effect but led to a reduced allergenicity in vitro, while we observed an adjuvant effect regarding IgG production in vivo.

  2. Expression and characterisation of a Psoroptes ovis glutathione S-transferase.

    Science.gov (United States)

    Lee, Alison J; Huntley, John; Van den Broek, Adri; Coates, David; Isaac, R Elwyn

    2002-04-19

    The astigmatid mite Psoroptes ovis is the causative agent of sheep scab, a highly contagious parasitic disease of sheep. Infection causes severe allergic dermatitis, resulting in damage to the fleece and hide, loss of condition and occasional mortality. Interest in the P. ovis allergens led us to characterise a glutathione S-transferase (GST) which displays homology to GST allergens isolated from the house dust mite, Dermatophagoides pteronyssinus and the cockroach, Blatella germanica. A cDNA encoding a mu-class GST from P. ovis was expressed in Escherichia coli and the recombinant protein purified for biochemical analysis. SDS-PAGE analysis indicated that the purified product was homogeneous and had an apparent molecular weight of 30 kDa. The recombinant GST (rGST) is active towards the substrate 1-chloro-2,4-dinitrobenzene (CDNB), whereas 1,2-dichloro-4-nitrobenzene (DCNB) is a poor substrate. The recombinant protein was also tested for recognition by IgE and IgG antibodies in serum from P. ovis naïve and P. ovis infested sheep. Neither IgE nor IgG antibodies were detected to the rGST. Prausnitz--Küstner testing with rGST did not provoke a characteristic weal and flare response. Biopsies collected at the PK test sites were stained for eosinophils, neutrophils, mast cells and basophils. Neutrophil, mast cell and basophil counts were not significantly different to the controls. Eosinophil numbers were significantly higher than controls, but were not due to an IgE response.

  3. 贝氏高原鳅脑垂体显微结构观察%Triplophysa bleekeri Pituitary Microstructure

    Institute of Scientific and Technical Information of China (English)

    刘小红; 何翔; 耿相昌; 李斌; 王志坚

    2009-01-01

    贝氏高原鳅的垂体由神经垂体和腺垂体构成,属于背腹型,但神经垂体稍偏向背部.腺垂体从前到后又依次分为前外侧部(RPD)、中外侧部(PPD)和中间部(PI).前外侧部由ACTH细胞和PRL细胞组成,中外侧部由GH细胞和大小两种嗜碱性细胞组成,中间部分布有MSH细胞和另一种嗜碱性细胞.神经垂体分枝丰富,遍布于腺垂体的各个部分,在中间部尤为丰富.垂体内血管发达.%According to histological and histochemical studies on the pituitary of Triplophysa bleekeri, the results show that the pituitary is composed of neurohypophysis and adenohypophysis, and is of the dorsal-ventral style. However the neurohypohysis is somewhat deviate to the dorsal part, the branches are abundant which are distributed in every part of the adenohypophysis, especially in the pars inermedia (PI). From the beginning to the end, the adenohypohhysis can be divided into the rostral pars distails (RPD), proximal pars distails (PPD) and pars intermedia (PI). There are corticotropes and lactotropes in the RPD, somatoropes, and 2 kinds of basophilic cells in the PPD, and melanotropes and also a basophilic cells in the PI. Blood vessels are abundant.

  4. Recombinant phospholipase A1 (Ves v 1 from yellow jacket venom for improved diagnosis of hymenoptera venom hypersensitivity

    Directory of Open Access Journals (Sweden)

    Grunwald Thomas

    2010-04-01

    Full Text Available Abstract Background Hymenoptera venoms are known to cause life-threatening IgE-mediated anaphylactic reactions in allergic individuals. Proper diagnosis of hymenoptera venom allergy using venom extracts is severely affected by molecular cross-reactivities. Although non-glycosylated marker allergens would facilitate the identification of the culprit venom, the major allergen phospholipase A1 (Ves v 1 from yellow jacket venom (YJV remained unavailable so far. Methods Expression of Ves v 1 as wild type and enzymatically inactivated mutant and Ves v 5 in insect cells yielded soluble proteins that were purified via affinity chromatography. Functionality of the recombinant allergens was assessed by enzymatic and biophysical analyses as well as basophil activation tests. Diagnostic relevance was addressed by ELISA-based analyses of sera of YJV-sensitized patients. Results Both major allergens Ves v 1 and Ves v 5 could be produced in insect cells in secreted soluble form. The recombinant proteins exhibited their particular biochemical and functional characteristics and were capable for activation of human basophils. Assessment of IgE reactivity of sera of YJV-sensitized and double-sensitized patients emphasised the relevance of Ves v 1 in hymenoptera venom allergy. In contrast to the use of singular molecules the combined use of both molecules enabled a reliable assignment of sensitisation to YJV for more than 90% of double-sensitised patients. Conclusions The recombinant availability of Ves v 1 from yellow jacket venom will contribute to a more detailed understanding of the molecular and allergological mechanisms of insect venoms and may provide a valuable tool for diagnostic and therapeutic approaches in hymenoptera venom allergy.

  5. Quantification of Anisakis simplex allergens in fresh, long-term frozen, and cooked fish muscle.

    Science.gov (United States)

    Rodríguez-Mahillo, Ana Isabel; González-Muñoz, Miguel; de las Heras, Cristina; Tejada, Margarita; Moneo, Ignacio

    2010-08-01

    Fish-borne parasitic zoonoses such as Anisakiasis were once limited to people living in countries where raw or undercooked fish is traditionally consumed. Nowadays, several factors, such as the growing international markets, the improved transportation systems, the population movements, and the expansion of ethnic ways of cooking in developed countries, have increased the population exposed to these parasites. Improved diagnosis technology and a better knowledge of the symptoms by clinicians have increased the Anisakiasis cases worldwide. Dietary recommendations to Anisakis-sensitized patients include the consumption of frozen or well-cooked fish, but these probably do not defend sensitized patients from allergen exposure. The aim of our work was to develop a sensitive and specific method to detect and quantify Anisakis simplex allergens in fish muscle and its derivatives. Protein extraction was made in saline buffer followed by preparation under acid conditions. A. simplex antigens were detected by IgG immunoblot and quantified by dot blot. The allergenic properties of the extracts were assessed by IgE immunoblotting and basophil activation test. We were able to detect less than 1 ppm of A. simplex antigens, among them the allergen Ani s 4, in fish muscle with no cross-reactions and with a recovery rate of 82.5%. A. simplex antigens were detected in hakes and anchovies but not in sardines, red mullets, or shellfish. We detected A. simplex allergens in cooked hakes and also in hake stock. We proved that A. simplex allergens are preserved in long-term frozen storage (-20 degrees C +/- 2 degrees C for 11 months) of parasitized hakes. Basophil activation tests have proven the capability of the A. simplex-positive fish extracts to induce allergic symptoms.

  6. A histamine release assay to identify sensitization to Culicoides allergens in horses with skin hypersensitivity.

    Science.gov (United States)

    Wagner, Bettina; Childs, Bronwen A; Erb, Hollis N

    2008-12-15

    Skin hypersensitivity is an allergic disease induced in horses by allergens of Culicoides midges. The condition is typically diagnosed by clinical signs and in some horses in combination with allergy testing such as intradermal skin testing or serological allergen-specific IgE determination. Here, we describe an alternative method for allergy testing: a histamine release assay (HRA) that combines the functional aspects of skin testing with the convenience of submitting a blood sample. The assay is based on the principle that crosslinking of allergen-specific IgE bound via high-affinity IgE receptors to the surfaces of mast cells and basophils induces the release of inflammatory mediators. One of these mediators is histamine. The histamine was then detected by a colorimetric enzyme-linked immunosorbent assay. The histamine assay was used to test 33 horses with skin hypersensitivity and 20 clinically healthy control animals for histamine release from their peripheral blood basophils after stimulation with Culicoides allergen extract or monoclonal anti-IgE antibody. An increased histamine release was observed in the horses with skin hypersensitivity compared to the control group after allergen-specific stimulation with Culicoides extract (p=0.023). In contrast, stimulation with anti-IgE induced similar amounts of released histamine in both groups (p=0.46). For further evaluation of the HRA, we prepared a receiver operating-characteristic (ROC) curve and performed a likelihood-ratio analysis for assay interpretation. Our results suggested that the assay is a valuable diagnostic tool to identify sensitization to Culicoides allergens in horses. Because some of the clinically healthy horses also showed sensitization to Culicoides extract, the assay cannot be used to distinguish allergic from non-allergic animals. The observation that sensitization is sometimes detectable in non-affected animals suggested that clinically healthy horses use immune mechanisms to control the

  7. Basaloid squamous cell carcinoma of the penis with papillary features: a clinicopathologic study of 12 cases.

    Science.gov (United States)

    Cubilla, Antonio L; Lloveras, Belén; Alemany, Laia; Alejo, María; Vidal, August; Kasamatsu, Elena; Clavero, Omar; Alvarado-Cabrero, Isabel; Lynch, Charles; Velasco-Alonso, Julio; Ferrera, Annabelle; Chaux, Alcides; Klaustermeier, Joellen; Quint, Wim; de Sanjosé, Silvia; Muñoz, Nubia; Bosch, Francisco Xavier

    2012-06-01

    There are 3 distinct variants of penile squamous cell carcinoma frequently associated with human papillomavirus (HPV): basaloid, warty-basaloid, and warty carcinomas. Considering the high incidence rates of penile cancer in some countries, a large international study was designed to evaluate the presence of HPV, its genotype distribution, and its association with histologic types of penile cancer. In this international review of >900 cases, we found a group of highly distinct papillary neoplasms composed of basophilic cells resembling urothelial tumors but frequently associated with HPV. Macroscopically, tumors were exophytic or exoendophytic. Microscopically, there was a papillomatous pattern of growth with a central fibrovascular core and small basophilic cells lining the papillae. Positivity for HPV was present in 11 of 12 tumors (92%). Single genotypes found were HPV-16 in 9 tumors and HPV-51 in 1 tumor. Multiple genotypes (HPV-16 and HPV-45) were present in another case. Overexpression of p16 was observed in all cases. Uroplakin-III was negative in all cases. The differential diagnosis was with basaloid, warty-basaloid, warty, and papillary squamous cell carcinoma and with urothelial carcinomas. Local excision (4 cases), circumcision (3 cases), or partial penectomy (5 cases) were preferred treatment choices. Tumor thickness ranged from 1 to 15 mm (average, 7 mm). Two patients with tumors invading 11 and 15 mm into the corpus spongiosum developed inguinal nodal metastasis. Of 11 patients followed up (median 48 mo), 7 were alive with no evidence of metastatic disease, 3 died from causes other than penile cancer, and another died postoperatively. This morphologically distinct tumor probably represents a papillary variant of basaloid carcinomas (papillary-basaloid carcinomas). Unlike typical basaloid carcinomas, the overall prognosis was excellent. However, deeply invasive tumors were associated with regional nodal metastasis indicating a potential for tumor

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

    Directory of Open Access Journals (Sweden)

    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.

  9. Human peripheral blood mononuclear cells exhibit heterogeneous CD52 expression levels and show differential sensitivity to alemtuzumab mediated cytolysis.

    Directory of Open Access Journals (Sweden)

    Sambasiva P Rao

    Full Text Available Alemtuzumab is a monoclonal antibody that targets cell surface CD52 and is effective in depleting lymphocytes by cytolytic effects in vivo. Although the cytolytic effects of alemtuzumab are dependent on the density of CD52 antigen on cells, there is scant information regarding the expression levels of CD52 on different cell types. In this study, CD52 expression was assessed on phenotypically distinct subsets of lymphoid and myeloid cells in peripheral blood mononuclear cells (PBMCs from normal donors. Results demonstrate that subsets of PBMCs express differing levels of CD52. Quantitative analysis showed that memory B cells and myeloid dendritic cells (mDCs display the highest number while natural killer (NK cells, plasmacytoid dendritic cells (pDCs and basophils have the lowest number of CD52 molecules per cell amongst lymphoid and myeloid cell populations respectively. Results of complement dependent cytolysis (CDC studies indicated that alemtuzumab mediated profound cytolytic effects on B and T cells with minimal effect on NK cells, basophils and pDCs, correlating with the density of CD52 on these cells. Interestingly, despite high CD52 levels, mDCs and monocytes were less susceptible to alemtuzumab-mediated CDC indicating that antigen density alone does not define susceptibility. Additional studies indicated that higher expression levels of complement inhibitory proteins (CIPs on these cells partially contributes to their resistance to alemtuzumab mediated CDC. These results indicate that alemtuzumab is most effective in depleting cells of the adaptive immune system while leaving innate immune cells relatively intact.

  10. Oral muscle relaxant may induce immediate allergic reactions.

    Science.gov (United States)

    Hur, Gyu-Young; Hwang, Eui Kyung; Moon, Jae-Young; Ye, Young-Min; Shim, Jae-Jeong; Park, Hae-Sim; Kang, Kyung-Ho

    2012-07-01

    Eperisone and afloqualone act by relaxing both skeletal and vascular smooth muscles to improve circulation and suppress pain reflex. These drugs are typically prescribed with non-steroidal anti-inflammatory drugs (NSAIDs) as painkillers. However, there have been no reports on serious adverse reactions to oral muscle relaxants; and this is the first report to describe three allergic reactions caused by eperisone and afloqualone. All three patients had histories of allergic reactions after oral intake of multiple painkillers, including oral muscle relaxants and NSAIDs, for chronic muscle pain. An open-label oral challenge test was performed with each drug to confirm which drugs caused the systemic reactions. All patients experienced the same reactions within one hour after oral intake of eperisone or afloqualone. The severity of these reactions ranged from laryngeal edema to hypotension. To confirm that the systemic reaction was caused by eperisone or afloqualone, skin prick testing and intradermal skin tests were performed with eperisone or afloqualone extract in vivo, and basophil activity tests were performed after stimulation with these drugs in vitro. In one patient with laryngeal edema, the intradermal test with afloqualone extract had a positive result, and CD63 expression levels on basophils increased in a dose-dependent manner by stimulation with afloqualone. We report three allergic reactions caused by oral muscle relaxants that might be mediated by non-immunoglobulin E-mediated responses. Since oral muscle relaxants such as eperisone and afloqualone are commonly prescribed for chronic muscle pain and can induce severe allergic reactions, we should prescribe them carefully.

  11. Identification, recombinant expression, and characterization of the 100 kDa high molecular weight Hymenoptera venom allergens Api m 5 and Ves v 3.

    Science.gov (United States)

    Blank, Simon; Seismann, Henning; Bockisch, Benjamin; Braren, Ingke; Cifuentes, Liliana; McIntyre, Mareike; Rühl, Dana; Ring, Johannes; Bredehorst, Reinhard; Ollert, Markus W; Grunwald, Thomas; Spillner, Edzard

    2010-05-01

    Insect stings can cause life-threatening IgE-mediated anaphylactic reactions in venom-allergic patients. Although several compounds have already been described as venom allergens, prominent allergen candidates especially in the higher m.w. range have still remained elusive. Tandem mass spectrometry-based sequencing assigned a candidate gene to the most prominent putative high m.w. allergen Api m 5 (allergen C) in honeybee (Apis mellifera) venom and also allowed identification of its homologue Ves v 3 in yellow jacket (Vespula vulgaris) venom. Both proteins exhibit a pronounced sequence identity to human dipeptidyl peptidase IV or CD26. Reactivity of a human IgE mAb verified the presence of these proteins in the venoms. Both proteins were produced in insect cells and characterized for their enzymatic activity as well as their allergenic potential using sera and basophils from insect venom-allergic patients. Both Api m 5 and Ves v 3 were recognized by specific IgE of the majority of patients even in the absence of cross-reactive carbohydrate determinants. Serologic IgE reactivity closely matched activation of human basophils by Api m 5 or Ves v 3, thus underlining their relevance in functional assays. With Api m 5 and Ves v 3, a new pair of homologous allergens becomes available for future clinical applications in diagnosis and therapy that may also contribute to the understanding of molecular mechanisms of insect venoms. Moreover, the patient IgE reactivity together with the cellular activation demonstrates for the first time the relevance of high m.w. allergens in the context of hymenoptera venom allergy.

  12. Mitochondrial N-formyl peptides induce cardiovascular collapse and sepsis-like syndrome.

    Science.gov (United States)

    Wenceslau, Camilla Ferreira; McCarthy, Cameron G; Szasz, Theodora; Goulopoulou, Styliani; Webb, R Clinton

    2015-04-01

    Fifty percent of trauma patients who present sepsis-like syndrome do not have bacterial infections. This condition is known as systemic inflammatory response syndrome (SIRS). A unifying factor of SIRS and sepsis is cardiovascular collapse. Trauma and severe blood loss cause the release of endogenous molecules known as damage-associated molecular patterns. Mitochondrial N-formyl peptides (F-MIT) are damage-associated molecular patterns that share similarities with bacterial N-formylated peptides and are potent immune system activators. The goal of this study was to investigate whether F-MIT trigger SIRS, including hypotension and vascular collapse via formyl peptide receptor (FPR) activation. We evaluated cardiovascular parameters in Wistar rats treated with FPR or histamine receptor antagonists and inhibitors of the nitric oxide pathway before and after F-MIT infusion. F-MIT, but not nonformylated peptides or mitochondrial DNA, induced severe hypotension via FPR activation and nitric oxide and histamine release. Moreover, F-MIT infusion induced hyperthermia, blood clotting, and increased vascular permeability. To evaluate the role of leukocytes in F-MIT-induced hypotension, neutrophil, basophil, or mast cells were depleted. Depletion of basophils, but not neutrophils or mast cells, abolished F-MIT-induced hypotension. Rats that underwent hemorrhagic shock increased plasma levels of mitochondrial formylated proteins associated with lung damage and antagonism of FPR ameliorated hemorrhagic shock-induced lung injury. Finally, F-MIT induced vasodilatation in isolated resistance arteries via FPR activation; however, F-MIT impaired endothelium-dependent relaxation in the presence of blood. These data suggest that F-MIT may be the link among trauma, SIRS, and cardiovascular collapse.

  13. 魁蚶血细胞分类及其免疫功能的初步分析%Studies on the hemocytes types and their immunological functions in bloody clam (Scapharca broughtonii)

    Institute of Scientific and Technical Information of China (English)

    周丽青; 杨爱国; 王清; 刘志鸿; 吴彪; 田吉腾; 严加坤; 吕振明

    2013-01-01

    借助显微观察和鳗弧菌免疫刺激等手段,首次对魁蚶血细胞进行分类及免疫功能分析.魁蚶血细胞可分为3大类:红细胞、白细胞和血栓细胞,红细胞发育过程大致经历新生、成熟、衰老和死亡4个阶段.白细胞分为:嗜酸性、嗜中性、嗜碱性细胞,以及淋巴细胞和巨核细胞.嗜酸性、嗜中性、嗜碱性细胞和血栓细胞均参与血液的凝固;红细胞、白细胞均有吞噬病原菌能力,红细胞具有被动吞噬作用和免疫吸附作用,白细胞具有主动吞噬作用,嗜酸性和嗜中性细胞是吞噬病原菌的主体,嗜碱性细胞具有吞噬大颗粒异物的作用,魁蚶感染病原菌后白细胞增多并表现炎症.研究表明,魁蚶血细胞在应对外界的免疫刺激时,能作出系统免疫应答,且血细胞免疫分工明确,以抵御外界不良刺激.%It was the first time to study the types and immunological functions of blood cells in bloody clam (Scapharca broughtonii) by the means of microscopic observation and immunostimulation with pathogenic Vibrio anguillarum, and three types of hemocytes were recognized: red blood cells, white blood cells and thrombus cells. Red blood cells had four developmental states: freshmen, mature, aging and death. White blood cells were divided into acidophils, neutuophils, basophils, lymphocytes, and megalokaryocytic. Acidophils,neutuophils,basophils and thrombus cells were all involved in the blood coagulation. Both red blood cells and white blood cells had phagocytic function,red cells showed the passive phagocytic function and immune adherence function, while white cells showed the active phagocytic function. Acidophils and neutuophils cells were principal in phagocytizing pathogens, and basophils cells could be found phagocytizing large size foreign materials, an increase in the number of white cell and inflammation took place after the bloody clam was infected by pathogen. It was also the first time to show the

  14. A lesson learnt: retrospection in a case of pilomatricoma mimicking as parotid neoplasm Uma lição aprendida: retrospectiva em um caso de pilomatricoma simulando neoplasia de parótida

    Energy Technology Data Exchange (ETDEWEB)

    Dutta, Mainak [Department of Otorhinolaryngology and Head-Neck Surgery, Medical College and Hospital, Kolkata (India); Chatterjee, Indranil [Department of Pediatric Surgery, Medical College and Hospital, Kolkata (India); Dutta, Mainak [Departamento de Otorrinolaringologia e Cirurgia de Cabeça e Pescoço, Medical College and Hospital, Kolkata (India); Chatterjee, Indranil [Departamento de Cirurgia Pediátrica, Medical College and Hospital, Kolkata (India)

    2016-07-01

    A 10-year-old girl presented with a painless, slow-growing swelling on left pre-auricular region that was noticed for 2 years (Figure 1). The swelling was bosselated, non-tender, firm-to-hard on palpation, had poorly-defined margin, and measured approximately 3.5cm x 3.0cm. In addition, it had restricted mobility, and seemed to be of parotid origin, with overlying skin apparently stretched and fixed. The ultrasonography was unable to delineate the depth and confirm the involvement of the parotid, although fine needle aspiration cytology (FNAC) suggested pleomorphic adenoma. For this reason, we decided to perform superficial parotidectomy. The histopathology of the lesion diagnosed pilomatricoma, which is an uncommon, benign ectodermal tumor of dermis/subdermis. Pilomatricoma constitutes a pluripotent cell expression in the germinal matrix center of hair follicles with differentiation into cortical cells, and the main relevance of its clinical presence is its potential to be misinterpreted, resulting in unnecessary aggressive interventions. It is often a diagnosis of retrospection. Some large series reported correct pre-operative diagnosis at 1.1 to 29%.{sup 1,2} A careful re-examination of pre-operative image revealed a subtle bluish tinge, which was not observed presumably because of the dark-skinned complexion, and the characteristic “tent sign”{sup 3}perceptible through the already-stretched and bosselated skin surface (Figure 1). Fine needle aspiration cytology is imperative for the assessment but is often erroneous{sup 4} due to inadequate, non-representative sampling from a lesion with complex cell contents. In a typical example, the basophilic basaloid cells with scanty cytoplasm and indistinct borders encase the anucleated cytoplasm-rich eosinophilic “ghost/shadow cells” (Figure 2) with areas of keratinization, giant-cells and dystrophic calcification in the matrix. As the tumor gets older, the central “ghost cells” increase at the expense of

  15. A macro- and light microscopical study of the pathology of Calicophoron microbothrium infection in experimentally infected cattle

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    M. Mavenyengwa

    2005-09-01

    Full Text Available Twelve Tuli weaner steers aged 1 year were randomly subdivided into three groups of four animals and infected with different doses of Calicophoron microbothrium metacercariae. Each animal in Group I received a low dose (LD of 5 000 metacercariae, Group II a medium dose (MD of 15 000 metacercariae, Group III a high dose (HD of 25 000 metacercariae and one additional animal was kept as an uninfected control (C. After infection, one animal from each group was slaughtered on Day 28, 42, 56 and 84 post infection (pi and samples from the ileum, jejunum, duodenum, abomasum and the rumen were collected for histopathological and cytological examination. On Day 28 pi, the gross pathological lesions observed in the duodenum of the LD and the MD animals were similar and comprised duodenal thickening, corrugation, hyperaemia, petechiation and ulceration. In the HD animal the duodenal lesions were similar but more severe. The abomasal folds were severely oedematous in the MD group and nearly occluded the abomasal lumen. Moderate oedema of the abomasal folds was also present in the LD and HD animals. The gross pathological lesions regressed in all the infected groups with increasing age of infection and had disappeared completely by Day 56 pi. On Day 28 pi the histopathological lesions in the duodenum and jejunum of the LD and MD groups were similar, comprising subtotal villous atrophy, hyperplasia of Brunner's glands and Peyer's patches and moderate infiltration of eosinophils, mast cells and a few globule leukocytes, basophils and lymphocytes in the lamina propria. The HD group had total villous atrophy, severe hyperplasia and cystic dilatation of Brunner's glands, which had expanded to cover the entire submucosa. On Day 42 pi the histopathological lesions were still present in the MD and the HD groups comprising subtotal villous atrophy and hyperplasia of Brunner's glands. Heavy infiltrations of eosinophils, moderate amounts of mast cells and a few

  16. A Human Lin(-) CD123(+) CD127(low) Population Endowed with ILC Features and Migratory Capabilities Contributes to Immunopathological Hallmarks of Psoriasis.

    Science.gov (United States)

    Mora-Velandia, Luz María; Castro-Escamilla, Octavio; Méndez, Andrés González; Aguilar-Flores, Cristina; Velázquez-Avila, Martha; Tussié-Luna, María Isabel; Téllez-Sosa, Juan; Maldonado-García, César; Jurado-Santacruz, Fermín; Ferat-Osorio, Eduardo; Martínez-Barnetche, Jesus; Pelayo, Rosana; Bonifaz, Laura C

    2017-01-01

    Innate lymphoid cells (ILC) are members of a heterogeneous family with a lymphoid origin that mimics the T helper (Th) cytokine profile. ILC are involved in early effector cytokine-mediated responses during infections in peripheral tissues. ILC also play an important role in chronic skin inflammatory diseases, including psoriasis. Although classical ILC express CD127, it has been recently reported that the presence of non-classical CD127(-) ILC populations and an early ILC precursor (EILP) CD127(low). ILC development has predominately been investigated in mouse models. However, in humans, different transcription factors have been described for ILC identification. NFIL3 (nuclear factor, IL-3 regulated) is crucial for ILC development in response to IL-7. CD123 (IL-3Rα) is usually used to exclude basophils during ILC identification, however, it is unknown if in response to IL-3, NFIL3 could be relevant to induce ILC features in Lin(-) CD123(+) populations in addition, is also unknown whether peripheral blood (PB) population with ILC features may have skin-homing potential to participate in skin inflammatory chronic diseases. Here, we report a Lin(-) CD123(+) CD127(low) CD7(+) CLA(+) population that share some phenotypic properties with basophils, but expresses several transcription factors for ILC commitment such as inhibitor of DNA binding 2 (Id2), NFIL3, promyelocytic leukemia zinc finger (PLZF), thymocyte selection-associated high-mobility group box protein (TOX), and T cell factor-1 (TCF-1). In addition, this population expresses different ILC markers: CD132, CD90, CD161, α4 integrin, c-Kit, CRTH2, AhR, and IL-23R. IL-3 prevents apoptosis and increases their NFIL3, TOX, and PLZF expression. In PB, the CD123(+) CD127(low) population is predominantly a conspicuous population that expresses T-bet and RORγt. The Lin(-) CD123(+) CD127(low) population in PB has a limited Th type cytokine expression and highly expresses IL-8. The Lin(-) CD123(+) CD127(low) population

  17. Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring Campaign 2003)

    Energy Technology Data Exchange (ETDEWEB)

    Marigomez, Ionan [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain)]. E-mail ionan.marigomez@ehu.es; Soto, Manu [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Cancio, Ibon [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Orbea, Amaia [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Garmendia, Larraitz [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Cajaraville, Miren P. [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain)

    2006-07-01

    In order to assess the biological effects of the Prestige oil spill (POS), mussels (Mytilus galloprovincialis), European hake (Merluccius merluccius) and European anchovy (Engraulis encrasicolus) were sampled between April and September 2003 in various geographical areas of Bay of Biscay: Galicia, Central Cantabrian and East Cantabrian. In mussels, several cell and tissue biomarkers were measured: peroxisome proliferation as induction of acyl-CoA oxidase (AOX) activity, lysosomal responses as changes in the structure (lysosomal volume density, V{sub V{sub L}}, surface-to-volume ratio, S/V{sub L}, and numerical density, N{sub V{sub L}}) and in membrane stability (labilization period, LP), cell type replacement as relative proportion of basophilic cells (volume density of basophilic cells, V{sub V{sub BAS}}) in digestive gland epithelium, and changes in the morphology of digestive alveoli as mean luminal radius to mean epithelial thickness (MLR/MET). Additionally, flesh condition index (FCI) and gonad index (GI) were measured as supporting parameters. In hake and anchovy, liver histopathology was examined to determine the prevalence of parasites, melanomacrophage centers, non-specific lesions (inflammatory changes, atrophy, necrosis, apoptosis), early non-neoplastic toxicopathic lesions (i.e., hepatocellular nuclear polymorphism), foci of cellular alteration, benign and malignant neoplasms. In mussels, AOX induction was noticeable in April except in Galicia. LP values were low in all the geographical areas studied, indicating disturbed health, especially in Galicia. Alike, lysosomal enlargement was observed in most stations as shown by the extremely low S/V{sub L} values. V{sub V{sub BAS}} and MLR/MET values were markedly high. Overall, employed biomarkers detected exposure to toxic chemicals and disturbed health in mussels from Bay of Biscay, with impact decreasing from April to September. Although hepatocellular nuclear polymorphism and nematode parasitization in

  18. Analysis of hematologic and serum chemistry values of Spheniscus magellanicus with molecular detection of avian malarial parasites (Plasmodium spp.

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    Sabrina D.E. Campos

    2014-12-01

    Full Text Available Magellanic penguins (Spheniscus magellanicus routinely migrate from their breeding colonies to Southern Brazil often contracting diseases during this migration, notably avian malaria, which has been already reported in Brazil and throughout the world. Detection of Plasmodium spp. in blood smears is the routine diagnostic method of avian malaria, however it has a low sensitivity rate when compared to molecular methods. Considering the negative impact of avian malaria on penguins, the aim of this study was to detect the presence of Plasmodium spp. in Magellanic penguins using Polymerase Chain Reaction (PCR and by verifying clinical, hematological, and biochemical alterations in blood samples as well as to verify the likely prognosis in response to infection. Blood samples were obtained from 75 penguins to determine packed cell volume (PCV, red blood cell (RBC and white blood cell (WBC counts, mean corpuscular volume (MCV, uric acid, total protein, albumin, globulin and aspartate aminotransferase (AST activity levels. Whole blood samples were used for PCR assays. Plasmodium spp. was detected in 32.0% of the specimens using PCR and in 29.3% using microscopic analyses. Anorexia, diarrhea and neurological disorders were more frequent in penguins with malaria and a significant weight difference between infected and non-infected penguins was detected. PCV and MCV rates showed no significant difference. RBC and WBC counts were lower in animals with avian malaria and leukopenia was present in some penguins. Basophil and lymphocyte counts were lower in infected penguins along with high monocyte counts. There was no significant difference in AST activities between infected and non-infected animals. There was a significant increase in uric acid values, however a decrease in albumin values was observed in infected penguins. Based on this study, we concluded that Plasmodium spp. occurs in Magellanic penguins of rehabilitation centers in Southeastern Brazil

  19. 论毛母质瘤临床病理联系%Pilomatricoma a clinicopathological analysis of 46 case

    Institute of Scientific and Technical Information of China (English)

    张崇辉

    2014-01-01

    Objective:To improve the level of awareness about the pilomatricoma by discussing and analyzing the clinical data and pathological performance of pilomatricoma.Methods:Retrospectivly analy-zing forty-six patients with pilomatricoma in our hospital from July 2007 to Aug.2014,discussing and analyzing the clinical data and pathological performance.Result:Pilomatricoma was more common in teens.The duration of the disease was from half a month to more than a decade.Heads,faces,necks and upper limbs were predilection sites of pilomatricoma.Shadow cel s were always found in histopatho-logic slides and basophilic cel s were found in most case.The basophilic cel s decreased with the extension of the course.Clinical y the misdiagnosis rate was very high.Conclusion:Pilomatricoma was a rare be-nign tumor.Pilomatricoma was more common in teens.Shadow cel s were its pathological feature and necessary to the correct diagnosis.%目的::通过探讨分析毛母质瘤的临床资料和病理表现等,提升对其认识水平。方法:对2007年7月~2014年8月于我院诊治的46例毛母质瘤患者进行回顾性分析,对其临床资料和病理表现进行探讨与分析。结果:毛母质瘤好发于青年,病程半月~十余年不等,头面颈部及上肢为其好发部位,病理组织切片中影细胞恒定出现,大多可见嗜碱细胞,随病程延长嗜碱细胞逐渐减少。结论:毛母质瘤是一种少见的良性肿瘤,好发于青年,影细胞是其病理特征及确诊依据。

  20. 皱纹盘鲍消化腺细胞类型和分泌产物%THE CELL TYPES AND SECRETION OF THE DIGESTIVE GLAND IN HALIOTIS DISCUS HANNAI INO

    Institute of Scientific and Technical Information of China (English)

    崔龙波; 刘传琳; 刘迅; 陆瑶华

    2001-01-01

    对皱纹盘鲍消化腺进行了组织学、组织化学、超微结构及酶活性测定等研究。消化腺由消化细胞和嗜碱性细胞组成。消化细胞能内吞腺管腔内的外源性物质,细胞内充满大量与异噬功能有关的囊泡。消化细胞具有内吞和细胞内消化、分泌、贮存等功能。嗜碱性细胞含有发达的粗面内质网以及许多含铁的折光小体,具有分泌和贮存金属离子的功能。消化腺还呈现多种水解酶活性。%Histology, histochemistry, ultrastructure and activity of some hydrolytical enzymes of the digestive gland of Haliotis discus hannai Ino have been studied.The gland consists of two types of cells, digestive cells and basophilic cells. The digestive cells are capable of ingesting exogenous material from the lumen of the tubule, and contain a number of vesicles concerning with the heterophagic function of the cells. The vesicles exhibit considerable variation in size and appearance in different regions of the cells. Those in the apical and sub-apical regions of the cells, which are considered as heterophagosomes, are involved in accumulation of the ingested material. Those in the mid-regions, regarded as heterolysosomes, give a positive reaction to tests for acid phosphatase and non-specific esterase. They concern with intracellular digestion of the ingested material. The vesicles in the basal regions, considered as residual bodies, are involved in accumulation of undigested and indigestible residues. The digestive cells secrete proteinase. Numerous lipid globules in the cells show that the digestive cells can reserve food material. The basophilic cells are characterized by the possession of a well-developed rough endoplasmic reticulum and many refractile spherules which give a intense reaction to test for iron. The cells also secrete proteinase. The refractile spherules which contain metals may serve a detoxification function. The tests for enzymatic activity in vitro show

  1. Chemokines: structure, receptors and functions. A new target for inflammation and asthma therapy?

    Directory of Open Access Journals (Sweden)

    F. A. A. van Acker

    1996-01-01

    Full Text Available Five to 10% of the human population have a disorder of the respiratory tract called ‘asthma’. It has been known as a potentially dangerous disease for over 2000 years, as it was already described by Hippocrates and recognized as a disease entity by Egyptian and Hebrew physicians. At the beginning of this decade, there has been a fundamental change in asthma management. The emphasis has shifted from symptom relief with bronchodilator therapies (e.g. β2-agonists to a much earlier introduction of anti-inflammatory treatment (e.g. corticosteroids. Asthma is now recognized to be a chronic inflammatory disease of the airways, involving various inflammatory cells and their mediators. Although asthma has been the subject of many investigations, the exact role of the different inflammatory cells has not been elucidated completely. Many suggestions have been made and several cells have been implicated in the pathogenesis of asthma, such as the eosinophils, the mast cells, the basophils and the lymphocytes. To date, however, the relative importance of these cells is not completely understood. The cell type predominantly found in the asthmatic lung is the eosinophil and the recruitment of these eosinophils can be seen as a characteristic of asthma. In recent years much attention is given to the role of the newly identified chemokines in asthma pathology. Chemokines are structurally and functionally related 8–10 kDa peptides that are the products of distinct genes clustered on human chromosomes 4 and 17 and can be found at sites of inflammation. They form a superfamily of proinflammatory mediators that promote the recruitment of various kinds of leukocytes and lymphocytes. The chemokine superfamily can be divided into three subgroups based on overall sequence homology. Although the chemokines have highly conserved amino acid sequences, each of the chemokines binds to and induces the chemotaxis of particular classes of white blood cells. Certain

  2. Características das células parasitadas pelo Trypanosoma cruzi na parede da veia central das supra-renais de Chagásicos crônicos

    Directory of Open Access Journals (Sweden)

    Hipolito de Oliveira Almeida

    1986-12-01

    Full Text Available Foram analisados os aspectos das células musculares parasitadas pelo Trypanosoma cruzi, na veia da supra-renal de chagásicos crônicos, através de exame ao microscópio óptico de lâminas coradas pela hematoxilina-eosina (HE, PAS, Feulgen e peroxidase-antiperoxidase (PAP para antigenos do T. cruzi. Além das modificações nucleares descritas anteriormente, os leiomiócitos parasitados exibem alterações citoplasmáticas que podem ser vistas mesmo em células que albergam poucos parasitas. As formas amastigotas geralmente estão envoltas por halo claro e o citoplasma restante adquire aspecto granuloso ou reticular, basófilopelo HE, sendo sempre PAS e Feulgen negativos. Estes dados sugerem que o material basófilo no citoplasma deve ser RNA ribossômico. A periferia dos ninhos que mostram uma "membrana" com reação do PAP para antigenos do T. cruzi fortemente positiva, poderia ser devida a reação cruzada de material celular rechaçado para a periferia ou a difusão de antigenos do T. cruzi e sua adsorção à periferia celular. O material citoplasmático PAP positivo poderia resultar de artefato, de reação imunocitoquimica cruzada, de antigenos tripanossomóticos difundidos ou de antigenos tripanossoma-simile resultantes de interações entre o leiomiócito e o parasita.Some morphologic aspects of the smooth muscle cells, parasitised by T. cruzi in the adrenal vein of chronic chagasic patients were studied. The staining techniques used were the following: Hematoxylin and Eosin (HE, PAS, Feulgen and the peroxidase antiperoxidase (PAP immunocyto chemical method for identification of T. cruzi antigen. The intracellular amastigotes were often surrounded by a clear halo and the cytoplasm of the parasitised smooth muscle cells were granular or reticular in appearance being basophil and, PAS and Feulgen negative. These data suggest that the cytoplasmatic basophil material could be ribosomic RNA. The intracellular nests of amastigotes were

  3. Evaluation of an anal sac adenocarcinoma tumor in a Spitz dog

    Institute of Scientific and Technical Information of China (English)

    Javad; Javanbakht; Abbas; Tavassoli; Atefeh; Sabbagh; Mehdy; Aghamohammmad; Hassan; Shohreh; Alian; Samakkhah; Radmehr; Shafiee; Ali; Lakzian; Vahideh; Rahmani; Ghalee; Sonia; Shoja; Gharebagh

    2013-01-01

    A 9-year-old emasculated male Spitz with tenesmus and constipation had a subcutaneous mass at the left ventral aspect of the anus with history of polyuria and polydipsia.A complete blood cell count,serum biochemistry panel,and urinalysis(cystocentesis sample)were evaluated.Abnormalities in the serum biochemistry panel included a mildly elevated serum cholesterol concentration(7.28 mmol/L;reference interval,2.70-5.94 mmol/L),increased serum alkaline phosphatase activity(184 U/L;reference interval,9-90 U/L),alanine transaminase(122 U/L;reference interval,5-60 U/L)activity and aspartate aminotransferase(80 U/L;reference interval,5-55 U/L)activity,severe increased total calcium concentration(16.3 mg/dL;reference interval,8.2-12.4 mg/dL or 9.3-11.4 mg/dL),and decreased total calcium concentration(3.4 mg/dL,reference interval,2.5-5.6mg/dL).Furthermore,testing revealed an increased intact parathyroid hormone concentration(38.6 pmol/L;reference interval,3-17 pmol/L).On cytologic and histopathologic examinations,various types of cells were observed.Most of the cells were oval to polygonal and had elliptical or elongate nuclei and a moderate amount of pale to basophilic cytoplasm.The remaining cells had round to oval nuclei and pale to basophilic cytoplasm.Cells of both types were loosely adhered to each other and were arranged in rosette-like structures.Both neoplastic cell types had fine homogenous chromatin and either a small indistinct nucleolus or no visible nucleolus.Mild anisokaryosis and anisocytosis were observed.Histologically,the mass consists of glandular structures formed by cuboidal cells admixed with bundles of spindle cells.Based on location and histologic features,the final diagnosis was adenocarcinoma of the apocrine gland of the anal sac,which should be included as a cytologic differential diagnosis when spindle cells and typical epithelial cells are observed in masses in the region of the anal sac of dogs.

  4. Evaluation of an anal sac adenocarcinoma tumor in a Spitz dog

    Institute of Scientific and Technical Information of China (English)

    Javad Javanbakht; Abbas Tavassoli; Atefeh Sabbagh; Mehdy Aghamohammmad Hassan; Shohreh Alian Samakkhah; Radmehr Shafiee; Ali Lakzian; Vahideh Rahmani Ghalee; Sonia Shoja Gharebagh

    2013-01-01

    A 9-year-old emasculated male Spitz with tenesmus and constipation had a subcutaneous mass at the left ventral aspect of the anus with history of polyuria and polydipsia. A complete blood cell count, serum biochemistry panel, and urinalysis (cystocentesis sample) were evaluated. Abnormalities in the serum biochemistry panel included a mildly elevated serum cholesterol concentration (7.28 mmol/L; reference interval, 2.70-5.94 mmol/L), increased serum alkaline phosphatase activity (184 U/L; reference interval, 9-90 U/L), alanine transaminase (122 U/L;reference interval, 5-60 U/L) activity and aspartate aminotransferase (80 U/L; reference interval, 5-55 U/L) activity, severe increased total calcium concentration (16.3 mg/dL; reference interval, 8.2-12.4 mg/dL or 9.3-11.4 mg/dL), and decreased total calcium concentration (3.4 mg/dL, reference interval, 2.5-5.6mg/dL). Furthermore, testing revealed an increased intact parathyroid hormone concentration (38.6 pmol/L; reference interval, 3-17 pmol/L). On cytologic and histopathologic examinations, various types of cells were observed. Most of the cells were oval to polygonal and had elliptical or elongate nuclei and a moderate amount of pale to basophilic cytoplasm. The remaining cells had round to oval nuclei and pale to basophilic cytoplasm. Cells of both types were loosely adhered to each other and were arranged in rosette-like structures. Both neoplastic cell types had fine homogenous chromatin and either a small indistinct nucleolus or no visible nucleolus. Mild anisokaryosis and anisocytosis were observed. Histologically, the mass consists of glandular structures formed by cuboidal cells admixed with bundles of spindle cells. Based on location and histologic features, the final diagnosis was adenocarcinoma of the apocrine gland of the anal sac, which should be included as a cytologic differential diagnosis when spindle cells and typical epithelial cells are observed in masses in the region of the anal sac of dogs.

  5. The aging heart.

    Science.gov (United States)

    Klausner, S C; Schwartz, A B

    1985-02-01

    Pathologic studies of the myocardium and valvular structures have failed to provide convincing evidence of gross or microscopic changes that can be ascribed to aging alone. Lipofuscin accumulation and basophilic degeneration in cardiac muscle cells appear to be the most consistent findings associated with aging, but they are found in other conditions. Without doubt, pathologic changes in the myocardium, valves, and coronary arteries are found more frequently in the hearts of elderly persons, but those changes are caused by disease processes associated with an aging population rather than the aging process itself. Both the sinus and atrioventricular nodes decrease in size with age owing to a loss of cellularity. These structures become infiltrated with collagen, elastic tissue, and reticular fibers. Some have found infiltration also with fat. Amyloid deposition, basophilic degeneration of cells, and lipofuscin accumulation occur but probably do not cause functional abnormalities. Similar but less dramatic changes occur in the bundle of His and individual bundle branches. Most of the data suggests that these aging changes are not due to vascular insufficiency. Age-related changes in intrinsic mechanical function have been identified as a prolongation of contraction duration, decreased inotropic responses to catecholamines and cardiac glycosides, and an increase in mechanical refractoriness. Other possible age-related changes include alterations in relaxation, which may or may not be independent of the prolongation of contraction, and changes in the viscoelastic properties of cardiac muscle. When examined in the context of the components of a model of excitation-contraction coupling, changes in action potential duration and the function of the sarcoplasmic reticulum emerge as the most likely bases for the changes. The electrical characteristics of sinus, atrioventricular, and His-Purkinje cells as well as atrial and ventricular muscle cells change with age. The sinus

  6. Waterborne cadmium impacts immunocytotoxic and cytogenotoxic endpoints in green-lipped mussel, Perna canaliculus

    Energy Technology Data Exchange (ETDEWEB)

    Chandurvelan, Rathishri, E-mail: rch118@uclive.ac.nz [School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8140 (New Zealand); Marsden, Islay D., E-mail: islay.marsden@canterbury.ac.nz [School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8140 (New Zealand); Gaw, Sally, E-mail: sally.gaw@canterbury.ac.nz [Department of Chemistry, University of Canterbury, Private Bag 4800, Christchurch 8140 (New Zealand); Glover, Chris N., E-mail: chris.glover@canterbury.ac.nz [School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8140 (New Zealand)

    2013-10-15

    Highlights: •Cadmium (Cd) significantly altered the proportional composition of haemocytes. •Subchronic exposure increased nuclear aberrations in gill cells. •Branchial nuclear aberrations correlated significantly with gill Cd accumulation. •Cd induced DNA damage in haemocytes. -- Abstract: Mussels are sentinel species that can be used to monitor coastal metal pollution through the application of biomarkers. Among the several important metal toxicants in coastal settings, cadmium (Cd) is of particular concern, being a non-essential metal that is known to cause harmful impacts in aquatic organisms at low concentrations. The aim of the present study was to examine the immunocytotoxic and cytogenotoxic effects of Cd on the green-lipped mussel, Perna canaliculus, under laboratory conditions. The acute (96 h; 0, 2000 and 4000 μg Cd L{sup −1}) and subchronic (28 d; 0, 200 and 2000 μg Cd L{sup −1}) toxic effects of waterborne Cd were measured in haemocytes and gill cells using differential haemocyte cell count, the micronucleus test and the comet assay. During subchronic exposure to Cd the relative counts of eosinophils and hyalinocytes increased significantly in Cd-exposed mussels while the proportion of basophils decreased. All of these effects were time- and concentration-dependent. Conversely, the relative numbers of basophils and eosinophils increased significantly during acute Cd exposure. Nuclear aberrations such as the formation of micronuclei, nuclear buds, fragmented-apoptotic cells and binuclei were observed in gill cells of Cd-exposed mussels. All of these parameters increased significantly at 2000 μg Cd L{sup −1} during subchronic exposure to Cd, and all showed a strong and significant correlation to gill Cd accumulation. Comet assay results demonstrated a significant increase in DNA damage in the haemocytes of mussels exposed to subchronic Cd concentrations. The results indicate that Cd has the capacity to induce immune system and genotoxic

  7. A Human Lin− CD123+ CD127low Population Endowed with ILC Features and Migratory Capabilities Contributes to Immunopathological Hallmarks of Psoriasis

    Science.gov (United States)

    Mora-Velandia, Luz María; Castro-Escamilla, Octavio; Méndez, Andrés González; Aguilar-Flores, Cristina; Velázquez-Avila, Martha; Tussié-Luna, María Isabel; Téllez-Sosa, Juan; Maldonado-García, César; Jurado-Santacruz, Fermín; Ferat-Osorio, Eduardo; Martínez-Barnetche, Jesus; Pelayo, Rosana; Bonifaz, Laura C.

    2017-01-01

    Innate lymphoid cells (ILC) are members of a heterogeneous family with a lymphoid origin that mimics the T helper (Th) cytokine profile. ILC are involved in early effector cytokine-mediated responses during infections in peripheral tissues. ILC also play an important role in chronic skin inflammatory diseases, including psoriasis. Although classical ILC express CD127, it has been recently reported that the presence of non-classical CD127− ILC populations and an early ILC precursor (EILP) CD127low. ILC development has predominately been investigated in mouse models. However, in humans, different transcription factors have been described for ILC identification. NFIL3 (nuclear factor, IL-3 regulated) is crucial for ILC development in response to IL-7. CD123 (IL-3Rα) is usually used to exclude basophils during ILC identification, however, it is unknown if in response to IL-3, NFIL3 could be relevant to induce ILC features in Lin− CD123+ populations in addition, is also unknown whether peripheral blood (PB) population with ILC features may have skin-homing potential to participate in skin inflammatory chronic diseases. Here, we report a Lin− CD123+ CD127low CD7+ CLA+ population that share some phenotypic properties with basophils, but expresses several transcription factors for ILC commitment such as inhibitor of DNA binding 2 (Id2), NFIL3, promyelocytic leukemia zinc finger (PLZF), thymocyte selection-associated high-mobility group box protein (TOX), and T cell factor-1 (TCF-1). In addition, this population expresses different ILC markers: CD132, CD90, CD161, α4 integrin, c-Kit, CRTH2, AhR, and IL-23R. IL-3 prevents apoptosis and increases their NFIL3, TOX, and PLZF expression. In PB, the CD123+ CD127low population is predominantly a conspicuous population that expresses T-bet and RORγt. The Lin− CD123+ CD127low population in PB has a limited Th type cytokine expression and highly expresses IL-8. The Lin− CD123+ CD127low population expresses skin

  8. Amniotic fluid stem cells rescue both in vitro and in vivo growth, innervation, and motility in nitrofen-exposed hypoplastic rat lungs through paracrine effects.

    Science.gov (United States)

    Pederiva, F; Ghionzoli, M; Pierro, A; De Coppi, P; Tovar, J A

    2013-01-01

    Lung hypoplasia can be prevented in vitro by retinoic acid (RA). Recent evidence suggests that amniotic fluid stem (AFS) cells may integrate injured lungs and influence their recovery. We tested the hypothesis that AFS cells might improve lung growth and motility by paracrine mechanisms. Pregnant rats received either nitrofen or vehicle on E9.5. In vitro E13 embryonic lungs were cultured in the presence of culture medium alone or with RA, basophils, or AFS cells. In vivo green fluorescent protein-expressing (GFP(+)) rat AFS cells were transplanted in nitrofen-exposed rats on E10.5. E13 lung explants were cultured before analysis. The surface, the number of terminal buds, and the frequency of bronchial contractions were assessed. Protein gene product 9.5 (PGP 9.5) and α-actin protein levels were measured. The lung explants transplanted with AFS cells were stained for α-actin, PGP 9.5, and TTF-1. The levels of FGF-10, VEGFα, and TGF-β1 secreted by the AFS cells in the culture medium were measured. Comparison between groups was made by ANOVA. In vitro, the surface, the number of terminal buds, and the bronchial peristalsis were increased in nitrofen+AFS cell explants in comparison with nitrofen-exposed lungs. While nitrofen+RA lungs were similar to nitrofen+AFS ones, basophils did not normalize these measurements. PGP 9.5 protein was decreased in nitrofen lungs, but after adding AFS cells, the value was similar to controls. No differences were found in the expression of α-actin. In vivo, the surface, number of terminal buds, and peristalsis were similar to control after injection of AFS cells in nitrofen-exposed rats. Colocalization with TTF-1-positive cells was found. The levels of FGF-10 and VEGFα were increased in nitrofen+AFS cell explants, while the levels of TGF-β1 were similar to controls. Lung growth, bronchial motility, and innervation were decreased in nitrofen explants and rescued by AFS cells both in vitro and in vivo, similarly to that observed

  9. Sequential univariate gating approach to study the effects of erythropoietin in murine bone marrow.

    Science.gov (United States)

    Achuthanandam, Ram; Quinn, John; Capocasale, Renold J; Bugelski, Peter J; Hrebien, Leonid; Kam, Moshe

    2008-08-01

    Analysis of multicolor flow cytometric data is traditionally based on the judgment of an expert, generally time consuming, sometimes incomplete and often subjective in nature. In this article, we investigate another statistical method using a Sequential Univariate Gating (SUG) algorithm to identify regions of interest between two groups of multivariate flow cytometric data. The metric used to differentiate between the groups of univariate distributions in SUG is the Kolmogorov-Smirnov distance (D) statistic. The performance of the algorithm is evaluated by applying it to a known three-color data set looking at activation of CD4+ and CD8+ lymphocytes with anti-CD3 antibody treatment and comparing the results to the expert analysis. The algorithm is then applied to a four-color data set used to study the effects of recombinant human erythropoietin (rHuEPO) on several murine bone marrow populations. SUG was used to identify regions of interest in the data and results compared to expert analysis and the current state-of-the-art statistical method, Frequency Difference Gating (FDG). Cluster analysis was then performed to identify subpopulations responding differently to rHuEPO. Expert analysis, SUG and FDG identified regions in the data that showed activation of CD4+ and CD8+ lymphocytes with anti-CD3 treatment. In the rHuEPO treated data sets, the expert and SUG identified a dose responsive expansion of only the erythroid precursor population. In contrast, FDG resulted in identification of regions of interest both in the erythroid precursors as well as in other bone marrow populations. Clustering within the regions of interest defined by SUG resulted in identification of four subpopulations of erythroid precursors that are morphologically distinct and show a differential response to rHuEPO treatment. Greatest expansion is seen in the basophilic and poly/orthochromic erythroblast populations with treatment. Identification of populations of interest can be performed

  10. Molecular detection of Ehrlichia canisand Anaplasma platys in dogs in Southern Brazil

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    Camila Serina Lasta

    Full Text Available The aims of this study were to determine the occurrence ofAnaplasma platys and Ehrlichia canisinfection in dogs in Porto Alegre, Southern Brazil; and to investigate their association with hematological abnormalities. Serum samples from 196 dogs were first tested using dot-ELISA for antibodies against Anaplasmaspp. and Ehrlichia canis. Peripheral blood samples from 199 dogs were subjected to 16S rRNA nested PCR (nPCR for A. platysand E. canis, followed by DNA sequencing to ensure pathogen identity. A total of 19/196 samples (9.69% were positive forAnaplasma spp. using ELISA and 28/199 (14.07% samples were positive for A. platys by nested PCR. All the dog samples were negative for E. canis, both in anti-E. canisantibody tests and in nested PCR. There were no significant differences in hematological parameters between A. platys-PCR positive and negative dogs and Anaplasma spp. serologically positive dogs, except for basophil counts, which were higher in nPCR-positive dogs. This is the first report showing A. platys presence in dogs in Southern Brazil. In conclusion, hematological parameters may not be sufficient to diagnose A. platys infection in dogs in Southern Brazil, probably due either to low pathogenicity or to chronic infection. On the other hand, E. canis may either have very low occurrence or be absent in dogs in Porto Alegre.

  11. Molecular detection of Ehrlichia canis and Anaplasma platys in dogs in Southern Brazil.

    Science.gov (United States)

    Lasta, Camila Serina; dos Santos, Andrea Pires; Messick, Joanne Belle; Oliveira, Simone Tostes; Biondo, Alexander Welker; Vieira, Rafael Felipe da Costa; Dalmolin, Magnus Larruscaim; González, Félix Hilario Diaz

    2013-01-01

    The aims of this study were to determine the occurrence of Anaplasma platys and Ehrlichia canis infection in dogs in Porto Alegre, Southern Brazil; and to investigate their association with hematological abnormalities. Serum samples from 196 dogs were first tested using dot-ELISA for antibodies against Anaplasma spp. and Ehrlichia canis. Peripheral blood samples from 199 dogs were subjected to 16S rRNA nested PCR (nPCR) for A. platys and E. canis, followed by DNA sequencing to ensure pathogen identity. A total of 19/196 samples (9.69%) were positive for Anaplasma spp. using ELISA and 28/199 (14.07%) samples were positive for A. platys by nested PCR. All the dog samples were negative for E. canis, both in anti-E. canis antibody tests and in nested PCR. There were no significant differences in hematological parameters between A. platys-PCR positive and negative dogs and Anaplasma spp. serologically positive dogs, except for basophil counts, which were higher in nPCR-positive dogs. This is the first report showing A. platys presence in dogs in Southern Brazil. In conclusion, hematological parameters may not be sufficient to diagnose A. platys infection in dogs in Southern Brazil, probably due either to low pathogenicity or to chronic infection. On the other hand, E. canis may either have very low occurrence or be absent in dogs in Porto Alegre.

  12. First case of Anaplasma platys infection in a dog from Croatia

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    Dyachenko Viktor

    2012-03-01

    Full Text Available Abstract Background It is known that Anaplasma (A. platys, the causative agent of infectious canine cyclic thrombocytopenia, is endemic in countries of the Mediterranean basin. However, few reports are available from the Balkans. This case report describes a dog, which was imported from Croatia to Germany in May 2010. One month later the dog was presented to a local veterinarian in Germany due to intermittent/recurrent diarrhoea. Diagnostic tests were performed to identify infections caused by Anaplasma spp., Ehrlichia spp., Hepatozoon canis, Babesia spp., Leishmania spp., Borrelia burgdorferi and/or Dirofilaria immitis. Findings Haematological examination of a blood smear revealed basophilic inclusions in thrombocytes, which were confirmed as A. platys with a species-specific real-time PCR. Additionally, an infection with Babesia (B. vogeli was also detected (PCR and serology. No specific antibodies against Anaplasma antigen were detectable. Although the dog showed no specific clinical signs, thrombocytopenia, anaemia and elevated C-reactive protein (CRP were observed. Sequencing of a 1,348-bp partial ribosomal RNA gene revealed highest homology to A. platys sequences from Thailand, Japan and France. Conclusions A. platys was detected for first time in a dog imported from Croatia. As the dog was also co-infected by B. vogeli, unique serological and haematological findings were recorded. Thrombocytopenia, anaemia and elevated values of C-reactive protein were the laboratory test abnormalities observed in this case. A. platys infections should be considered in dogs coming from Croatia and adjacent regions.

  13. Plasmoblastic lymphoma associated with human immunodeficiency virus.

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    Horváth, Emoke; Krenács, L; Bagdi, Eniko; Pávai, Z; Macarie, I; Nagy, Elod-Erno; Demian, Smaranda

    2008-01-01

    Plasmoblastic lymphoma (PBL) is a subtype of the diffuse large B-cell lymphoma, typically present as extranodal disease associated with human immune deficiency virus (HIV) infection. PBLs are often the initial manifestation of AIDS. Here we present a case of PBL concerning the oral cavity. A 34-year-old woman presented a tumor in the oral cavity that involved the maxilla and gingiva (confirmed by CT-scan). The gingival biopsy showed a massive infiltration by large lymphoid cells with round, vesicular nuclei, prominent nucleoli, fine chromatin and an significant amount of basophilic cytoplasm which express CD79a, CD138, cytoplasmic lambda light chain and LCA, without staining for CD20, CD38, CD3 and CTK. Serological analysis confirmed HIV positivity. PBLs lack most B-lineage markers, but many express CD79a in at least some of the cells, therefore generate difficulties in differential diagnosis. Overall assessment and correlation of the histopathological and immunohistochemical features with the clinical findings and serology investigation are the most helpful diagnostic tools and can lead to the final diagnosis.

  14. Autologous serum skin test as an indicator of chronic autoimmune urticaria in a tertiary care hospital in South India

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    Adaikalampillai Ganapathy Vikramkumar

    2014-01-01

    Full Text Available Background: Autologous serum skin test (ASST is a simple in-vivo clinical test for the detection of basophil histamine releasing activity and to diagnose chronic autoimmune urticaria (CAU among chronic spontaneous urticaria (CSU patients. Diagnosing these patients is also important as they may need high doses of antihistamines and systemic corticosteroids during acute exacerbations. Aims and Objectives: The aim of this study is to study the prevalence of CAU among cases of CSU by using ASST. Materials and Methods: This was a cross-sectional study done among 48 patients presenting with CSU. Detailed history, physical examination and routine investigations were recorded for all patients. ASST was done on all the 48 patients. Results: Of the 48 patients included in the study, 20 patients (41.6% were ASST positive, while the remaining 28 (58% were ASST negative. The median duration of disease in both ASST positive and negative patients was 1 year. ASST positivity was higher (66.6% among patients with a history of round shaped weals, though not statistically significant. ASST positivity was seen in 5 (71.4% out of seven patients with systemic involvement, which was again not statistically significant. Conclusion: Our study did not show any significant difference between patients with and without antibodies regarding mean age and sex distribution, clinical morphology of individual weals, duration, severity, systemic symptoms, angioedema, atopy, and association with other autoimmune conditions.

  15. Structure and Function of the Digestive System of Solen Grandis Dunker

    Institute of Scientific and Technical Information of China (English)

    SHENG Xiuzhen; ZHAN Wenbin; REN Sulian

    2003-01-01

    Structure and function of the digestive system of a bivalve mollusc, Solen grandis, were studied using light microscopy and histochemical methods. The wall of digestive tube consists of four layers: the mucosal epithelium, connective tissue, muscular and fibrosa or serosa (only in the portion of rectum) from the inner to the outer. The ciliated columnar epithelial cells, dispersed by cup-shaped mucous cells, rest on a thin base membrane. There are abundant blood spaces in connective tissue layer. The digestive diverticula are composed of multi-branched duct and digestive tubules. The digestive tubules are lined with digestive and basophilic secretory cells, and surrounded by a layer of smooth muscle fibers and connective tissues. Activities of acid and alkaline phosphatases, esterase and lipase are detected in the digestive cells, and the epithelia of stomach and intestine, suggesting that these cells are capable of intracellular digesting of food materials and absorbing.Besides, acid phosphatase and esterase activities are present in the posterior portion of esophagus. Phagocytes are abundant in blood spaces and the lumens of stomach and intestine, containing brown granules derived from the engulfed food materials. The present work indicates that phagocytes play important roles in ingestion and digestion of food materials, which is supported as well by the activities of acid phosphatase, esterase and lipase detected in blood spaces.

  16. Enhancement of third harmonic contrast with harmonophores in multimodal non-linear microscopy of histological sections

    Science.gov (United States)

    Tuer, Adam; Bakueva, Ludmilla; Cisek, Richard; Alami, Jennifer; Dumont, Daniel J.; Rowlands, John; Barzda, Virginijus

    2008-02-01

    Histological investigations of biological tissue benefited tremendously from staining different cellular structures with various organic dyes. With the introduction of new imaging modalities such as second harmonic generation (SHG) and third harmonic generation (THG) microscopy, the demand for novel dyes that enhance the harmonic signals has arisen. The new labels with high molecular hyperpolarizability have recently been termed harmonophores. In this study, we demonstrate that hematoxylin, the standard histological stain used in H&E (hematoxylin and eosin) staining, enhances the microscopic THG signal. Hematoxylin has an affinity for basophilic structures such as the cell nucleus, ribosomes and mitochondria, while eosin stains structures such as the cytoplasm, collagen and red blood cells. The histological sections of H&E stained cancerous prostate tissue found in transgenic adenocarcinoma of the mouse prostate (TRAMP) have been investigated with the multimodal SHG, THG and multiphoton excitation fluorescence (MPF) microscope. Strong THG signal revealed intracellular structures originating where the hematoxylin stain resides, while SHG imaging of the tissue showed the presence of collagen fibrils in the extracellular matrix. The MPF was mostly present in the extracellular matrix. The spectrally and temporally resolved MPF revealed that most of the fluorescence originates from the eosin. The THG image did not correlate with MPF confirming that the harmonic signal originates from hematoxylin. Multimodal nonlinear microscopy adds invaluable information about cellular structures to the widely used bright field investigations of H&E stained histological sections, and can be efficiently used for morphological studies as well as cancer diagnostics.

  17. Disseminated histoplasmosis (Histoplasma capsulatum) in a pet rabbit: case report and review of the literature.

    Science.gov (United States)

    Brandão, João; Woods, Samantha; Fowlkes, Natalie; Leissinger, Mary; Blair, Robert; Pucheu-Haston, Cherie; Johnson, James; Elster Phillips, Christina; Tully, Thomas

    2014-01-01

    A 2.5-year-old intact male miniature lop rabbit (Oryctolagus cuniculus) was presented with multiple nodules surrounding the eyes, nose, mouth, and prepuce. Cytological evaluation of the periocular nodules revealed the presence of intracellular (within macrophages) and extracellular yeast organisms. The yeast organisms were approximately 3-5 µm in diameter, round to oval, with a thin clear capsule, and contained an eccentrically placed basophilic crescent-shaped nucleus. The clinical pathological interpretation was granulomatous inflammation with intralesional yeast of a morphology consistent with Histoplasma spp. The rabbit was treated with microsized griseofulvin (25 mg/kg, orally, once a day) for 12 days pending final cytological diagnosis of histoplasmosis. No significant improvement was noted during the treatment period, and humane euthanasia was performed. Postmortem examination revealed the presence of intracellular and extracellular yeast organisms in the small intestine, skin (antebrachium, perioral, palpebral, perianal, and pinnal), penis, penile urethra, rectum, axillary lymph node, and conjunctiva. Postmortem fungal culture yielded Histoplasma capsulatum. Based on clinical and postmortem findings, a definitive diagnosis of disseminated histoplasmosis was made. Disseminated histoplasmosis appears to be unreported in rabbits. Although the treatment used did not provide noticeable improvement, available information on histoplasmosis treatment in other species has been reviewed to provide useful information for future management of this condition in rabbits.

  18. The Research Progress of Myeloid Cells Development and Differentiation%髓系细胞发育分化的研究进展

    Institute of Scientific and Technical Information of China (English)

    周庆; 郝璐(综述); 蔡志明; 贝锦新(审校)

    2016-01-01

    The differentiation of myeloid cells is very important to the establishment of immune system . Under the control of specific growth factors and cytokines, bone marrow hematopoietic stem cells could develop into the myeloblasts and further into the mature granulocytes .According to the molecular weight and cytospin staining,granulocytes can be divided into eosinophils,neutrophils and basophils.In recent years,sig-nificant breakthroughs have been made in the classification definition and developmental differentiation of myeloid cells,providing references for the diagnosis and treatment of blood and immune system diseases.%髓系细胞发育分化对于免疫系统的形成至关重要。在特定生长因子和细胞因子的调控下,骨髓造血干细胞先发育为原粒细胞,进而发育为成熟的粒细胞。根据粒细胞分子量大小和染色情况,可以分为嗜酸粒细胞,中性粒细胞和嗜碱粒细胞。近年来,髓系细胞发育过程的分级定义及其发育调控过程的研究取得了巨大进步,为血液及免疫系统疾病的诊治提供了参考。

  19. Immunocalization of telomerase in cells of lizard tail after amputation suggests cell activation for tail regeneration.

    Science.gov (United States)

    Alibardi, L

    2016-02-01

    Tail amputation (autotomy) in most lizards elicits a remarkable regenerative response leading to a new although simplified tail. No information on the trigger mechanism following wounding is known but cells from the stump initiate to proliferate and form a regenerative blastema. The present study shows that telomerases are mainly activated in the nuclei of various connective and muscle satellite cells of the stump, and in other tissues, probably responding to the wound signals. Western blotting detection also indicates that telomerase positive bands increases in the regenerating blastema in comparison to the normal tail. Light and ultrastructural immunocytochemistry localization of telomerase shows that 4-14 days post-amputation in lizards immunopositive nuclei of sparse cells located among the wounded tissues are accumulating into the forming blastema. These cells mainly include fibroblasts and fat cells of the connective tissue and satellite cells of muscles. Also some immature basophilic and polychromatophilic erytroblasts, lymphoblasts and myelocytes present within the Bone Marrow of the vertebrae show telomerase localization in their nuclei, but their contribution to the formation of the regenerative blastema remains undetermined. The study proposes that one of the initial mechanisms triggering cell proliferation for the formation of the blastema in lizards involve gene activation for the production of telomerase that stimulates the following signaling pathways for cell division and migration.

  20. Enzymatic Hydrolysis Does Not Reduce the Biological Reactivity of Soybean Proteins for All Allergic Subjects.

    Science.gov (United States)

    Panda, Rakhi; Tetteh, Afua O; Pramod, Siddanakoppalu N; Goodman, Richard E

    2015-11-04

    Many soybean protein products are processed by enzymatic hydrolysis to attain desirable functional food properties or in some cases to reduce allergenicity. However, few studies have investigated the effects of enzymatic hydrolysis on the allergenicity of soybean products. In this study the allergenicity of soybean protein isolates (SPI) hydrolyzed by Alcalase, trypsin, chymotrypsin, bromelain, or papain was evaluated by IgE immunoblots using eight soybean-allergic patient sera. The biological relevance of IgE binding was evaluated by a functional assay using a humanized rat basophilic leukemia (hRBL) cell line and serum from one subject. Results indicated that hydrolysis of SPI by the enzymes did not reduce the allergenicity, and hydrolysis by chymotrypsin or bromelain has the potential to increase the allergenicity of SPI. Two-dimensional (2D) immunoblot and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of the chymotrypsin-hydrolyzed samples indicated fragments of β-conglycinin protein are responsible for the apparent higher allergenic potential of digested SPI.

  1. Recent advances in the understanding of the penicillin urticarias.

    Science.gov (United States)

    Jillson, O F; Porter, P S

    1965-08-01

    The three major groups of immunoglobulins (gamma G, gamma A, and gamma M) associated with this disease are reviewed. The presence or absence of atopic disease may account for percentage variability of gamma A because reagins (skin-sensitizing antibodies) are found in this immunoglobulin. The gamma A is the antibody usually responsible for anaphylaxis, rather than the gamma G precipitins, so stressed in the past. All three immunoglobulins may be found in serum sickness, which could account for the complex nature of this type of penicillin urticaria. The merits of the immunological tests (penicilloyl-polylysine, benzyl penicillin, hemagglutination, basophil degranulation) for the detection of penicillin sensitivity are analyzed, particularly as each applies to the various types of penicillin urticaria (serum sickness, anaphylactic, dermographic, delayed dermographic, and simple chronic urticaria and the lupus diathesis). The penicilloyl-polylysine test is greatly overrated as a means of predicting possible anaphylaxis. The benzyl penicillin skin test properly performed is an excellent means of indicating this.

  2. T Cell Epitope Immunotherapy Induces a CD4+ T Cell Population with Regulatory Activity

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    Verhoef Adrienne

    2005-01-01

    Full Text Available Background Synthetic peptides, representing CD4+ T cell epitopes, derived from the primary sequence of allergen molecules have been used to down-regulate allergic inflammation in sensitised individuals. Treatment of allergic diseases with peptides may offer substantial advantages over treatment with native allergen molecules because of the reduced potential for cross-linking IgE bound to the surface of mast cells and basophils. Methods and Findings In this study we address the mechanism of action of peptide immunotherapy (PIT in cat-allergic, asthmatic patients. Cell-division-tracking dyes, cell-mixing experiments, surface phenotyping, and cytokine measurements were used to investigate immunomodulation in peripheral blood mononuclear cells (PBMCs after therapy. Proliferative responses of PBMCs to allergen extract were significantly reduced after PIT. This was associated with modified cytokine profiles generally characterised by an increase in interleukin-10 and a decrease in interleukin-5 production. CD4+ cells isolated after PIT were able to actively suppress allergen-specific proliferative responses of pretreatment CD4neg PBMCs in co-culture experiments. PIT was associated with a significant increase in surface expression of CD5 on both CD4+ and CD8+ PBMCs. Conclusion This study provides evidence for the induction of a population of CD4+ T cells with suppressor/regulatory activity following PIT. Furthermore, up-regulation of cell surface levels of CD5 may contribute to reduced reactivity to allergen.

  3. Acute hepatopancreatic necrosis disease (AHPND) outbreaks in Penaeus vannamei and P. monodon cultured in the Philippines.

    Science.gov (United States)

    de la Peña, Leobert D; Cabillon, Nikko Alvin R; Catedral, Demy D; Amar, Edgar C; Usero, Roselyn C; Monotilla, Wilberto D; Calpe, Adelaida T; Fernandez, Dalisay Dg; Saloma, Cynthia P

    2015-10-27

    Acute hepatopancreatic necrosis disease (AHPND) has recently emerged as a serious disease of cultured shrimp. It has also been described as early mortality syndrome (EMS) due to mass mortalities occurring within 20 to 30 d after stocking of ponds with postlarvae. Here, Penaeus vannamei and Penaeus monodon from shrimp farms in the Philippines were examined for the toxin-producing strain of Vibrio parahaemolyticus due to AHPND-like symptoms occurring in marketable size shrimp. In the P. vannamei, histology revealed typical AHPND pathology, such as sloughing of undifferentiated cells in the hepatopancreatic tubule epithelium. Analysis using the IQ2000 AHPND/EMS Toxin 1 PCR test generated 218 bp and 432 bp amplicons confirmative of the toxin-producing strain of V. parahaemolyticus among shrimp sampled from 8 of 9 ponds. In the P. monodon, histology revealed massive sloughing of undifferentiated cells of the hepatopancreatic tubule epithelium in the absence of basophilic bacterial cells. PCR testing generated the 2 amplicons confirmatory for AHPND among shrimp sampled from 5 of 7 ponds. This study confirms the presence of AHPND in P. vannamei and P. monodon farmed in the Philippines and suggests that the disease can also impact late-stage juvenile shrimp.

  4. COLEUS (PLECTRANTHUS BARBATUS – A MULTIPURPOSE MEDICINAL HERB

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    SharmaYashaswini

    2011-03-01

    Full Text Available Plectranthus barbatus Andr. (Syn. Coleus forskohlii Briq. is a perennial herb, belonging to the family Lamiaceae. Its tuberous roots are found to be a rich source of forskohlin (coleonol used as a potential drug for hypertension, congestive heart failure, eczema, colic, respiratory disorders, painful urination, insomnia, and convulsions. Clinical studies of the plant further support these traditional uses, indicating therapeutic benefit in asthma, angina, psoriasis and prevention of cancer metastases. Forskolin directly activates almost all hormone sensitive adenylate cyclases in intact cells, tissues and even solubilised preparation of adenylate cyclase. Stimulation of adenylate cyclase is thought to be the mechanism by which forskolin relaxes a variety of smooth muscles. Forskolin, by increasing cAMP level in turn, inhibits basophil and mast cell degranulation and histamine release, lowers blood pressure and intraocular pressure and it inhibits platelet aggregation, promotes vasodilation, bronchodilation, and thyroid hormone secretion. Coleus acts as a natural source of drug for many major diseases implying that there is a great demand for production and processing of the crop. The paper deals with botany, medicinal uses, phytochemistry, mechanism of action and case studies on coleus.

  5. Histopathologic changes of hypothalamus and pituitary in a rat model of polycystic ovary syndrome

    Institute of Scientific and Technical Information of China (English)

    Jiang Yan; Meng Fan-yu; Hu Zhen-hua; Liu Fang

    2010-01-01

    Objective: To investigate the histopathologic changes of hypothalamus and pituitary in a rat model of polycystic ovary syndrome (PCOS).Methods: Rat model of PCOS was established in 6 immature female SD rats and another 6 immature rats treated with placebo were as control. The tissues of hypothalamus and pituitary were obtained and observed by light microscope and transmission electron microscope.Results: Light microscopy revealed little difference in morphology of neurons in arcuate nucleus or basophilic cells in pituitary between PCOS rats and normal rats. Electron microscopic examination showed that, compared with those in normal rats, GnRH neurons in PCOS rats were larger and fuller, with Golgi complex and mitochondria increased. The mitochondria were small, round and swelling. More high-density secretory granules and bright vesicles were observed in the cytoplasm. The Golgi complex near nucleus in pituitary gonadotropin cell in PCOS rats was fractured and expanded, and there were increased number mitochondria and different sizes of the higher electron density secretory granules in the cytoplasm. Conclusion: The morphological alterations in hypothalamus and pituitary could play a very important role in the development of PCOS.

  6. Burkitt Lymphoma Presenting as Unilateral Deafness in an Immunocompetent Patient

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    Andre Pinto

    2012-01-01

    Full Text Available A 55-year-old HIV-negative white male presented with right ear deafness, right axillary lymphadenopathy, and weight loss. Laboratory findings included anemia, marked leukocytosis, and thrombocytopenia. Examination of the peripheral smear demonstrated the presence of increased circulating blast-like cells of intermediate size, with basophilic cytoplasm and nuclei with open chromatin. MRI of the brain was compatible with hemorrhagic labyrinthitis. Excisional biopsy of the axillary mass revealed an enlarged lymph node with effaced architecture and “starry sky” appearance. The cells expressed CD20, CD10, BCL6, and surface kappa immunoglobulin light chain, with a high proliferative index by immunohistochemistry and flow cytometry. Subsequent bone marrow biopsy was hypercellular (approximately 95%, with blast-like cells virtually replacing all hematopoietic elements. Routine karyotype as well as FISH analysis of bone marrow cells demonstrated rearrangement of the MYC gene at chromosome 8q24 region, IGH/MYC fusion, and additional signal for IGH gene. We present herein a case of sporadic Burkitt lymphoma occurring in a previously healthy HIV-negative male. The unusual clinical findings in this case include the relatively older age at presentation (55 years, an immunocompetent patient who had nodal involvement and leukemic phase of Burkitt, coupled with partial deafness. A brief educational review of this neoplasm is made.

  7. Food allergy to the carbohydrate galactose-alpha-1,3-galactose (alpha-gal): four case reports and a review.

    Science.gov (United States)

    Bircher, Andreas J; Hofmeier, Kathrin Scherer; Link, Susanne; Heijnen, Ingmar

    2017-02-01

    Until recently, food allergies to mammalian meats have been considered to be very rare. The observation that patients not previously exposed to the monoclonal chimeric antibody cetuximab suffered from severe anaphylaxis upon first exposure, led to the identification of galactose-alpha-1,3-galactose as a new relevant carbohydrate allergen. These patients later often suffered from anaphylactic reactions to red meat. Epidemiological data indicated that bites by the tick Amblyomma americanum in the USA, later also by Ixodes species in other continents, resulted in sensitisation to alpha-gal. On the other hand, in African patients with parasitic disorders, a high prevalence of anti-alpha-gal IgE, without clinical relevance, has been reported. In our four cases, one patient with a late onset of meat allergy had a history of a tick bite. The other three patients had symptoms from childhood or at a juvenile age. This indicates that in some patients, other ways of sensitisation may also take place. However, in patients without atopy, tick bite-induced IgE to alpha-gal may be more relevant. Diagnosis is based on a history of delayed onset of anaphylaxis. Skin tests with commercially available meat test solutions are often equivocal or negative; skin tests with raw meat and particularly pork kidney are more sensitive. Determination of specific IgE to alpha-gal is commercially available. The highest sensitivity is observed with skin and basophil activation tests with cetuximab which is, however, limited by its high costs.

  8. Role of Interleukin-33 in Innate-Type Immune Cells in Allergy

    Directory of Open Access Journals (Sweden)

    Susumu Nakae

    2013-01-01

    Full Text Available Interleukin-33 (IL-33, a member of the IL-1 cytokine family, is preferentially and constitutively expressed in epithelial cells, and it is especially localized in the cells' nucleus. The nuclear IL-33 is released by necrotic cells after tissue injury and/or trauma, and subsequently provokes local inflammation as an alarmin, like high-mobility group box protein-1 (HMGB-1 and IL-1α. IL-33 mainly activates Th2 cells and such innate-type immune cells as mast cells, basophils, eosinophils and natural helper cells that express IL-33R (a heterodimer of IL-1 receptor-like 1 [IL-1RL1; also called ST2, T1, Der4, fit-1] and IL-1 receptor accessory protein [IL-1RAcP]. That activation causes the cells to produce Th2 cytokines, which contribute to host defense against nematodes. On the other hand, excessive and/or inappropriate production of IL-33 is also considered to be involved in the development of such disorders as allergy. In this review, we summarize current knowledge regarding the pathogenic roles of IL-33 in the development of allergic inflammation by focusing on its effects on innate-type immune cells.

  9. Rickettsia-like organism infection in a freshwater cultured fish Ophiocephalus argus C.in China

    Institute of Scientific and Technical Information of China (English)

    GUO Qionglin; JIA Weizhang; HAN Xianpu; CAI Taozhen; GONG Xiaoning; SUN Xiaofeng

    2004-01-01

    From 2001 to 2002,a new and emergent infectious disease of Ophiocephalus argus occurred in a fishery in Hubei Province,China,with an incidence of 60%~70% and a mortality as high as 100%.The diseased fish showed an enlarged abdomen,the millet-like nodules in internal organs,and the swollen kidney which was composed of 5~10 sarcoma-like bodies in cream or gray-white colour or ulcerated into beandregs-like substance.Light microscopic observation revealed the basophilic or acidphilic inclusions in cytoplasm of the cells and the granulomas,a diffusive chronic inflammation in internal organs.Further analysis under an electron microscope indicated that the intracytoplasmic inclusions were rickettsia-like organisms (RLOs) that are either spherical or coccoid,with variable size,ranging from 0.5~1.5 μm in diameter,and enclosed within membrane-bound cytoplasmic vacuoles.RLO had a central nucleoid region with some fine filamentous structures and an electron-dense granule.Its cytoplasm contained abundant ribosomal bodies.Occasionally,RLO appeared to be divided by binary fission.RLOs were also observed in the homogenized tissue of infected fish.The results suggested that the death of cultured O.Argus was caused by RLO infection.

  10. Investigation of drugs responsible for perioperative anaphylactic reactions using cellular allergen stimulation test

    Institute of Scientific and Technical Information of China (English)

    Xin Xin; Zou Yi; Xing Lijiao; Yin Jia; Gu Jianqing; Wang Zixi; Huang Yuguang

    2014-01-01

    Background Anaphylactic reactions during anesthesia and operation are common and life threatening.Follow-up investigation is necessary for avoiding potential re-exposure of the patients to the offending drugs.The purpose of this study was to assess cellular allergen stimulation test (CAST) as a diagnostic instrument in immunoglobulin E (IgE)-and non-lgE-mediated anaphylactic reactions.Methods This study included 25 patients who developed perioperative anaphylactic reactions and 10 subjects that tolerated anesthetics and other drugs during perioperative period from September 2009 to October 2013 in Peking Union Medical College Hospital.We performed skin tests and flow cytometric analysis of basophil activation-based CAST in all subjects.Results Of the 25 patients,17 had IgE-mediated anaphylactic reactions (causative agent identified by skin tests) and 8 had non-lgE-mediated anaphylactic reactions (negative skin tests).CAST showed a sensitivity of 42.9%,specificity of 90%,and negative predictive value of 80.6% for neuromuscular blocking agents.Conclusions CAST may be useful for the diagnosis of anaphylactic reactions during perioperative period.Our findings call for further investigation to increase the sensitivity of the test.

  11. Changes of hematological references depends on storage period and temperature conditions in rats and dogs

    Science.gov (United States)

    Lee, Jung-Min

    2016-01-01

    Because changes in rat and dog hematological parameters according to storage conditions have been poorly documented, we sought to examine such changes. Blood analysis was performed using two hematology analyzers (ADVIA 2120i and Sysmex XN-V) after storage at room temperature and in cold storage for 5, 24, and 48 h, respectively. Interassay coefficients of variation for hematological parameters analyzed with the ADVIA 2120i and the XN-V showed similar. The levels of hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and platelet (PLT) showed significant variations with time in blood samples of rats and dogs. The leukocyte subpopulation showed high variation with storage conditions. The data for leukocyte differential counts obtained using the ADVIA 2120i, XN-V, and a manual differential counting procedure showed good agreement for neutrophils and lymphocyte counts, but monocytes, eosinophils, and basophils showed differences between the procedures. In conclusions, most rat and dog hematological parameters showed minimal changes; however, some showed high variation with storage time and temperature, especially PLT and leukocyte subpopulations. In conclusion, when performing hematological analysis in dogs and rats, it will be exactitude to analyze blood samples in fresh condition and at least within 24 h in the cold storage. PMID:28053618

  12. Correlating cell morphology and stochastic gene expression using fluorescence spectroscopy and GPU-enabled image analysis

    Science.gov (United States)

    Shepherd, Douglas; Shapiro, Evan; Perillo, Evan; Werner, James

    2014-03-01

    Biological processes at the microscopic level appear stochastic, requiring precise measurement and analytical techniques to determine the nature of the underlying regulatory networks. Single-molecule, single-cell studies of gene expression have provided insights into how cells respond to external stimuli. Recent work has suggested that macroscopic cell properties, such as cell morphology, are correlated with gene expression. Here we present single-cell studies of a signal-activated gene network: Interleukin 4 (IL4) RNA production in rat basophil leukemia (RBL) cells during the allergic response. We fluorescently label individual IL4 RNA transcripts in populations of RBL cells, subject to varying external stimuli. A custom super-resolution microscope is used to measure the number of fluorescent labeled IL4 transcripts in populations of RBL cells on a cell-by-cell basis. To test the hypothesis that cell morphology is connected genotype, we analyze white light images of RBL cells and cross-reference cell morphology with IL4 RNA levels. We find that the activation of RBL cells, determined by white-light imaging, is well correlated with IL4 mRNA expression.

  13. 1株湿地稀有放线菌的多相分类鉴定%Polyphasic Taxonomy of a Rare Marine Actinomycetes

    Institute of Scientific and Technical Information of China (English)

    唐树戈; 张柳; 于基成; 刘秋; 齐小辉

    2014-01-01

    Strain S402003 was isolated from marine sediment collected from Yalu River in Dandong, Liaoning Province. Based on the polyphasic studies, including the morphology, physiological and biochemical characteristics, chemotaxonomy and 16S rRNA gene sequence analysis, the results showed that strain S402003 had lipase activity and was capable to reduce nitrate, belongs to the mild salt-tolerant and basophilic bacteria. It was primarily identified as Brevibacterium linens with the similarity 98.834%.%从鸭绿江滨海湿地样品中分离得到1株稀有海洋放线菌S402003,通过形态观察、培养特征、生理生化特征、化学组分鉴定以及16S rRNA基因序列分析对其进行多相分类鉴定。结果表明:该菌株具有脂酶活性,能还原硝酸盐,属于轻度耐盐、嗜碱菌。该菌株属于Brevibacterium linens,相似性为98.834%。

  14. Mast Cells and IgE: From History to Today

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    Hirohisa Saito

    2013-01-01

    Full Text Available Role of mast cells in allergy had remained undetermined until the discovery of IgE in 1966. Then, IgE purified from many Liters of plasma, which had been donated from a patient with fatal myeloma, was distributed to researchers all over the world, and thus accelerated exploring the mechanisms involved in allergic reactions, particularly about the role of mast cells and basophils in the IgE-mediated reactions. Identification of mast cells as a progeny of a bone marrow hematopoietic stem cell in 1977 led us to successful in vitro culture of human mast cells. Along with the development of molecular biological techniques, the structure of the high affinity IgE receptor (FceRI was determined in 1989. These findings and subsequent investigations brought deeper understanding of IgE-mediated allergic diseases in the past half century, especially where mast cells are involved. We have now even obtained the information about whole genome expression of FceRI-dependently activated mast cells. In sharp contrast to our comprehension of allergic diseases where IgE and mast cells are involved, the mechanisms involved in non-IgE-mediated allergic diseases or non-IgE-mediated phase of IgE-mediated diseases are almost left unsolved and are waiting for devoted investigators to reveal it.

  15. Immunohistochemical identification of mast cells in formaldehyde-fixed tissue using monoclonal antibodies specific for tryptase.

    Science.gov (United States)

    Walls, A F; Jones, D B; Williams, J H; Church, M K; Holgate, S T

    1990-10-01

    An avidin-biotin enhanced immunoperoxidase procedure using monoclonal antibodies (AA1, AA3, and AA5) prepared against human mast cell tryptase resulted in intense staining of mast cells in paraffin-embedded tissue. The distribution of mast cells observed was similar to that seen when adjacent serial sections were stained using a standard procedure with toluidine blue, though the immunoperoxidase technique permitted the identification of significantly more mast cells. With monoclonal antibody AA1, immunostaining was entirely specific for mast cell granules, and there was negligible background staining in a range of tissues including lung, tonsil, colon, gastric mucosa, skin, and pituitary. There was no staining of antibody on basophils or on any other normal blood leukocyte. The technique was effective with tissue fixed in either Carnoy's or neutral buffered formalin, though the internal mast cell structure was better preserved with formaldehyde fixation. The immunoperoxidase staining procedure with monoclonal antibody AA1 is a highly specific and sensitive means for the detection of mast cells in routinely processed tissues.

  16. The impact of the 67kDa laminin receptor on both cell-surface binding and anti-allergic action of tea catechins.

    Science.gov (United States)

    Fujimura, Yoshinori; Umeda, Daisuke; Yamada, Koji; Tachibana, Hirofumi

    2008-08-15

    Here, we investigated the structure-activity relationship of major green tea catechins and their corresponding epimers on cell-surface binding and inhibitory effect on histamine release. Galloylated catechins; (-)-epigallocatechin-3-O-gallate (EGCG), (-)-gallocatechin-3-O-gallate (GCG), (-)-epicatechin-3-O-gallate (ECG), and (-)-catechin-3-O-gallate (CG) showed the cell-surface binding to the human basophilic KU812 cells by surface plasmon resonance analysis, but their non-galloylated forms did not. Binding activities of pyrogallol-type catechins (EGCG and GCG) were higher than those of catechol-type catechins (ECG and CG). These patterns were also observed in their inhibitory effects on histamine release. Previously, we have reported that biological activities of EGCG are mediated through the binding to the cell-surface 67kDa laminin receptor (67LR). Downregulation of 67LR expression caused a reduction of both activities of galloylated catechins. These results suggest that both the galloyl moiety and the B-ring hydroxylation pattern contribute to the exertion of biological activities of tea catechins and their 67LR-dependencies.

  17. THE HISTOLOGICAL CHANGES OF REPRODUCTIVE SYSTEM BETWEEN PRE-AND POST SPAWNING ONCHIDIUM STRUMA%瘤背石磺产卵前后生殖系统的组织学变化

    Institute of Scientific and Technical Information of China (English)

    吴旭干; 胡冰; 杨筱珍; 成永旭

    2009-01-01

    conducted to study the composition of its reproductive system, and no publication could be found on the histological changes of reproductive system during the spawning process of 0. struma. So, the present experiments were conducted to investigate the histological changes of reproductive system between pre- and post spawning 0. struma. After that, the composition and function of reproduction system (gonads and accessory sex organs) were further discussed in this paper, which should provide valuable information to understand reproductive process and improve hatchery technology of 0. struma. The results indicated that: (1) The male reproductive system included penis, penial sac, male prostate, digenetic gonad (mainly produce sperm in early stage of gonad development) and sperm bursa, while female reproductive system was consisted of digenetic gonad (mainly produce mature oocytes in late stage of gonad development) , gametal transportation tract, egg albumen gland, mucous gland, fertilization chamber and vagina; (2) Although there was no significant difference of male reproductive system between pre- and post spawning 0. struma, the significant differences could be found on the digenetic gland, egg albumen gland and mucous gland; (3) After spawning, the digenetic gland became loose while the un-spawned mature oocyte and the early previtellogeic oocyte were found in some follicles; (4) Before spawning, the egg albumen gland consisted of many small basophilic granules which structure was very similar to the outer albumen of the embryo of 0. struma. However, after spawning, the egg albumen gland contained bigger acidophilic granules than those before spawning; (5) Before spawning, there were acidophilic, basophilic and mixed area in the histological section of mucous gland while the basophilic area were dominant in the post-spawning mucous gland and the basophilic substance could be found in its secretion tube. In conclusion, these results suggested that the egg albumen gland

  18. Histopathological changes in the head kidney induced by cadmium in a neotropical fish Colossoma macropomum

    Directory of Open Access Journals (Sweden)

    R. Salazar-Lugo

    2013-12-01

    Full Text Available We evaluated the effect of cadmium (Cd on the structure and function of the head kidney in the freshwater fish Colossoma macropomum (C. macropomum. Juveniles were exposed to 0.1 mg/L CdCl2 for 31 days. Blood samples were examined using hematological tests and head kidney histology was determined by light microscopy. The concentration of Cd in the head and trunk kidneys was measured using an atomic absorption spectrophotometer. Cd produced histopathological changes in the head kidney, the most evident of these being: the thickening of the vein wall, an increase in the number of basophils/mast cells close to blood vessels and a severe depletion of hematopoietic precursors especially the granulopoietic series. In the blood, a decrease in the total leucocytes and hemoglobin concentration was observed. Cd-exposed fish showed higher Cd concentrations in the trunk kidney than the head kidney. In conclusion, exposure to Cd affected precursor hematopoietic cells in C. macropomum.

  19. Preparation and identification of Per a 5 as a novel American cockroach allergen.

    Science.gov (United States)

    Wei, Ji-Fu; Yang, Haiwei; Li, Dongning; Gao, Peisong; He, Shaoheng

    2014-01-01

    Glutathione S-transferase (GST) from various arthropods can elicit allergic reactions. In the present study, Per a 5, a GST, was cloned from American cockroach (CR) and expressed in both baculovirus-infected insect cell (iPer a 5) and E. coli expression (bPer a 5) systems. The secondary structures were predicted to be 45.93 and 8.69% of α-helix β-sheets in iPer a 5 and 42.54 and 8.49% of α-helix and β-sheets in bPer a 5, respectively. It is found that 4 out of 16 (25%) sera from American CR allergy patients reacted to both bPer a 9 and iPer a 9 as assessed by ELISA and Western blotting analysis, confirming that Per a 5 is not a major allergen of American CR. Induction of upregulated expression of CD63 and CCR3 on passively sensitized human basophils (sera from American CR allergy patients) by approximately up to 4.5- and 3.2-fold indicates that iPer a 5 and bPer a 5 are functionally active. Recombinant Per a 5 (rPer a 5) should be a useful tool for studying and understanding the role of Per a 5 in CR allergy.

  20. [The high-affinity IgE receptor: lessons from structural analysis].

    Science.gov (United States)

    Blank, Ulrich; Jouvin, Marie-Hélène; Guérin-Marchand, Claudine; Kinet, Jean-Pierre

    2003-01-01

    The high affinity receptor for IgE, FcERI, is at the core of the allergic reaction. This receptor is expressed mainly on mast cells and basophils. Interaction of an allergen with its specific IgE bound to FcERI triggers cell activation, which induces the release of numerous mediators that are responsible for allergic manifestations. The recent increase in the prevalence of allergic diseases in developed countries has resulted in renewed efforts towards the development of new drugs. One of these is a humanised antibody directed against the IgE ligand. This antibody recognises specifically free but not FcERI-bound IgE thus preventing ligand binding and subsequent cell activation. This antibody has shown some efficacy in clinical trials involving patients with asthma and allergic rhinitis. The recent elucidation of the tridimensional structure of the complex between IgE and FcERI provides unexpected information regarding the mechanism of assembly of the complex, which now can be used to design small chemical compounds capable of specifically inhibiting this interaction.