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Zeitschriftenartikel zum Thema „CD antigens“

Autor: Grafiati

Veröffentlicht am 4. Juni 2021

Zuletzt aktualisiert am 30. Juli 2024

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1

Woolfson, Adrian, Justin Stebbing, Brian D. M. Tom, Kerryn J. Stoner, Walter R. Gilks, David P. Kreil, Stephen P. Mulligan et al. „Conservation of unique cell-surface CD antigen mosaics in HIV-1–infected individuals“. Blood 106, Nr. 3 (01.08.2005): 1003–7. http://dx.doi.org/10.1182/blood-2004-12-4642.

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AbstractCluster of differentiation (CD) antigens are expressed on cells of myeloid and lymphoid lineages. As most disease processes involve immune system activation or suppression, these antigens offer unique opportunities for monitoring host responses. Immunophenotyping using limited numbers of CD antigens enables differentiation states of immune system cells to be determined. Extended phenotyping involving parallel measurement of multiple CD antigens may help identify expression pattern signatures associated with specific disease states. To explore this possibility we have made a CD monoclonal antibody array and scanner, enabling the parallel immunophenotyping of leukocyte cell suspensions in a single and rapid analysis. To demonstrate this approach, we used the specific example of patients infected with human immunodeficiency virus type-1 (HIV-1). An invariant HIV-induced CD antigen signature has been defined that is both robust and independent of clinical outcome, composed of a unique profile of CD antigen expression levels that are both increased and decreased relative to internal controls. The results indicate that HIV-induced changes in CD antigen expression are disease specific and independent of outcome. Their invariant nature indicates an irreversible component to retroviral infection and suggests the utility of CD antigen expression patterns in other disease settings.

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2

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD antigens 2001“. Journal of Leukocyte Biology 70, Nr. 5 (November 2001): 685–90. http://dx.doi.org/10.1189/jlb.70.5.685.

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3

Kishimoto, T., S. Goyert, H. Kikutani, D. Mason, M. Miyasaka, L. Moretta, T. Ohno et al. „CD Antigens 1996“. Blood 89, Nr. 10 (15.05.1997): 3502. http://dx.doi.org/10.1182/blood.v89.10.3502.

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4

Kishimoto, T., S. Goyert, H. Kikutani, D. Mason, M. Miyasaka, L. Moretta, T. Ohno et al. „CD Antigens 1996“. Blood 89, Nr. 10 (15.05.1997): 3502. http://dx.doi.org/10.1182/blood.v89.10.3502.3502_3502_3502.

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5

Schlossman, SF, L. Boumsell, W. Gilks, JM Harlan, T. Kishimoto, C. Morimoto, J. Ritz, S. Shaw, RL Silverstein und TA Springer. „CD antigens 1993“. Blood 83, Nr. 4 (15.02.1994): 879–80. http://dx.doi.org/10.1182/blood.v83.4.879.879.

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6

Schlossman, SF, L. Boumsell, W. Gilks, JM Harlan, T. Kishimoto, C. Morimoto, J. Ritz, S. Shaw, RL Silverstein und TA Springer. „CD antigens 1993“. Blood 83, Nr. 4 (15.02.1994): 879–80. http://dx.doi.org/10.1182/blood.v83.4.879.bloodjournal834879.

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7

Schlossman, S. F., L. Boumsell, W. Gilks, J. M. Harlan, T. Kishimoto, C. Morimoto, J. Ritz et al. „CD Antigens 1993“. Leukemia & Lymphoma 13, sup1 (Januar 1994): 59–60. http://dx.doi.org/10.3109/10428199409052676.

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8

Mason, D. Y., P. André, A. Bensussan, C. Buckley, C. Civin, E. Clark, M. De Haas et al. „CD antigens 2001“. Tissue Antigens 58, Nr. 6 (Dezember 2001): 425–30. http://dx.doi.org/10.1034/j.1399-0039.2001.580614.x.

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9

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD Antigens 2001“. Modern Pathology 15, Nr. 1 (Januar 2002): 71–76. http://dx.doi.org/10.1038/modpathol.3880492.

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10

Knapp, W., B. Dorken, P. Rieber, RE Schmidt, H. Stein und AE von dem Borne. „CD antigens 1989“. Blood 74, Nr. 4 (01.09.1989): 1448–50. http://dx.doi.org/10.1182/blood.v74.4.1448.1448.

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11

Knapp, W., B. Dorken, P. Rieber, RE Schmidt, H. Stein und AE von dem Borne. „CD antigens 1989“. Blood 74, Nr. 4 (01.09.1989): 1448–50. http://dx.doi.org/10.1182/blood.v74.4.1448.bloodjournal7441448.

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12

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD antigens 2001“. International Immunology 13, Nr. 9 (September 2001): 1095–98. http://dx.doi.org/10.1093/intimm/13.9.1095.

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13

Schlossman, S. F., L. Boumsell, W. Gilks, J. M. Harlan, T. Kishimoto, C. Morimoto, J. Ritz et al. „CD antigens 1993“. Immunology Today 15, Nr. 3 (März 1994): 98–99. http://dx.doi.org/10.1016/0167-5699(94)90149-x.

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14

Mason, David, Pascale Andre, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD antigens 2001“. Immunology 103, Nr. 4 (August 2001): 401–6. http://dx.doi.org/10.1046/j.1365-2567.2001.01295.x.

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15

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD antigens 2002“. Blood 99, Nr. 10 (15.05.2002): 3877–80. http://dx.doi.org/10.1182/blood.v99.10.3877.

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16

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD Antigens 2001“. Cellular Immunology 211, Nr. 2 (August 2001): 81–85. http://dx.doi.org/10.1006/cimm.2001.1831.

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17

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „CD antigens 2001“. European Journal of Immunology 31, Nr. 10 (Oktober 2001): 2841–47. http://dx.doi.org/10.1002/1521-4141(2001010)31:10<2841::aid-immu2841>3.0.co;2-y.

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18

Knapp, W., B. Dörken, P. Rieber, R. E. Schmidt, H. Stein und A. E. G. Kr Von Dem Borne. „CD Antigens 1989“. International Journal of Cancer 44, Nr. 1 (15.07.1989): 190–91. http://dx.doi.org/10.1002/ijc.2910440135.

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19

Schlossman, S. F., L. Boumsell, W. Gilks, J. M. Harlan, T. Kishimoto, C. Morimoto, J. Ritz, S. Shaw, R. L. Silverstein und T. A. Springer. „CD antigens 1993.“ Journal of Immunology 152, Nr. 1 (01.01.1994): 1–2. http://dx.doi.org/10.4049/jimmunol.152.1.1.

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20

Hanafiah, Alfizah, Asif Sukri, Nik Ritza Kosai, Mohamad Aznan Shuhaili, Mustafa Mohammed Taher und Raja Affendi Raja Ali. „Immunophenotyping of Gastritis, Gastric Ulcer and Gastric Cancer using a Cluster of Differentiation (CD) Antibody Microarray“. Sains Malaysiana 52, Nr. 1 (31.01.2023): 187–97. http://dx.doi.org/10.17576/jsm-2023-5201-15.

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One of the factors that contribute to the development of gastric cancer is the host immune response. Extensive immunophenotype of gastric cancer can be identified by using antibody microarray technique that profiles more than 100 cluster of differentiation (CD) antigens in parallel. In this study, we used DotScanTM antibody microarray to profile CD antigen expression in patients with distinct digestive diseases for surface antigen disease-signatures. Patients’ blood samples with gastric disorders and healthy controls were taken and processed for leukocytes isolation using Histopaque density gradient centrifugation. Leukocytes were captured onto DotScanTM slides and cell binding densities were analyzed using DotReaderTM. Different groups of gastric diseases were found to be characterized by differentially expressed distinct CD antigens. Compared to normal healthy controls, 17, two and four highly expressed CD antigens were identified in gastritis, gastric ulcer and gastric cancer patients, respectively. The 17 CD antigens that show differential expression in gastritis were CD15, CD16, CD20, CD23, CD24, CD25, CD28, CD34, CD37, CD77, CD102, CD103, CD122, CD128, CD10, CD183, and CD184. High expression of CD64 and CD69 were found in gastric ulcer, whereas CD52, CD126, CD135, and CD121a were highly expressed in gastric cancer. CD antigens involve in T-cell functions had reduced expression in gastric cancer, while proinflammatory cytokines shows increased expression. These results demonstrate specific immunophenotype of CD antigens in patients with various gastric diseases and identification of differential expressed surface antigens may have clinical significance for diagnostic and therapeutic purposes.

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21

Simon, M., J. Antalíková, Ľ. Horovská, J. Jankovičová, K. Fábryová, S. Hluchý, P. Chrenek und V. Tančin. „Analysis of rabbit cell surface (CD) antigens by means of cross-reactive monoclonal antibodies with specificity for cattle CD antigens“. Czech Journal of Animal Science 54, No. 6 (22.06.2009): 270–76. http://dx.doi.org/10.17221/1728-cjas.

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Studies that involved testing monoclonal antibodies (mAbs) for cross-species reactivity proved to be efficient for the identification of previously unrecognized antigens in a number of different species. Twenty-six mAbs specific to different bovine CD (cluster defined) antigens (CD9, CD18, CD45R, CD41/61, CD62L, MHC class I and bovine IgG light chain molecule) were assayed for reactivity with rabbit peripheral blood leukocytes. Four of the mAbs recognizing CD9 and CD41/61 were reactive with rabbit platelets or granulocytes. These were investigated further by immunoblotting and immunohistochemical staining. The study identified CD9 and CD41/61 molecules on rabbit cells by mAbs IVA-50 and IVA-38. It showed that IVA-50 is a new valuable CD9 reagent for rabbit immunology which could be used for immunofluorescence staining or ELISA assay, immunohistological and molecular studies of rabbit CD9 antigen. IVA-38 recognizes the CD41/61 on rabbit platelets in indirect immunofluorescence and ELISA assay.

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22

Mason, David, Pascale André, Armand Bensussan, Chris Buckley, Curt Civin, Edward Clark, Masja de Haas et al. „Reference: CD Antigens 2002“. Journal of Immunology 168, Nr. 5 (01.03.2002): 2083–86. http://dx.doi.org/10.4049/jimmunol.168.5.2083.

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23

Rodríguez Sillke, Y., M. Schumann, D. Lissner, F. Branchi, R. Glauben und B. Siegmund. „P043 Small intestinal inflammation but not colitis drives pro-inflammatory nutritional antigen-specific T-cell response“. Journal of Crohn's and Colitis 14, Supplement_1 (Januar 2020): S154—S155. http://dx.doi.org/10.1093/ecco-jcc/jjz203.172.

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Abstract Background Inflammatory bowel disease (IBD) represents a dysregulation of the mucosal immune system. The pathogenesis of Crohn’s disease (CD) and ulcerative colitis (UC) is linked to the loss of intestinal tolerance and barrier function. The healthy mucosal immune system has previously been shown to be inert against food antigens. Since the small intestine is the main contact surface for antigens and therefore the immunological response, the present study served to analyse food-antigen-specific T cells in the peripheral blood of IBD patients. Methods Peripheral blood mononuclear cells of CD, with an affected small intestine, and UC (colitis) patients, either active or in remission, were stimulated with the following food antigens: gluten, soybean, peanut and ovalbumin. Healthy controls and celiac disease patients were included as controls. Antigen-activated CD4+ T cells in the peripheral blood were analysed by a magnetic enrichment of CD154+ effector T cells and a cytometric antigen-reactive T-cell analysis (‘ARTE’ technology) followed by characterisation of the effector response. Results The effector T-cell response of antigen-specific T cells were compared between CD with small intestinal inflammation and UC where inflammation was restricted to the colon. Among all tested food antigens, the highest frequency of antigen-specific T cells (CD4+CD154+) was found for gluten. Celiac disease patients were included as control, since gluten has been identified as the disease-causing antigen. The highest frequency of gluten antigen-specific T cells was revealed in active CD when compared with UC, celiac disease on a gluten-free diet (GFD) and healthy controls. Ovalbumin-specific T cells were almost undetectable, whereas the reaction to soybean and peanut was slightly higher. But again, the strongest reaction was observed in CD with small intestinal involvement compared with UC. Remarkably, in celiac disease on a GFD only antigen-specific cells for gluten were detected. These gluten-specific T cells were characterised by up-regulation of the pro-inflammatory cytokines IFN-γ, IL-17A and TNF-α. IFN-g was exclusively elevated in CD patients with active disease. Gluten-specific T-cells expressing IL-17A were increased in all IBD patients. Furthermore, T cells of CD patients, independent of disease activity, revealed a high expression of the pro-inflammatory cytokine TNF-α. Conclusion The ‘ARTE’-technique allows to analyse and quantify food antigen specific T cells in the peripheral blood of IBD patients indicating a potential therapeutic insight. These data provide evidence that small intestinal inflammation in CD is key for the development of a systemic pro-inflammatory effector T-cell response driven by food antigens.

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24

Hudson, AM, V. Makrynikola, A. Kabral und KF Bradstock. „Immunophenotypic analysis of clonogenic cells in acute lymphoblastic leukemia using an in vitro colony assay“. Blood 74, Nr. 6 (01.11.1989): 2112–20. http://dx.doi.org/10.1182/blood.v74.6.2112.2112.

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Abstract A culture system was used to analyze the expression of membrane differentiation antigens on the proliferative or clonogenic fraction of cells from cases of common (precursor B) acute lymphoblastic leukemia (c-ALL). Colonies of leukemic cells were obtained in 18 of 20 cases after 1 week in culture in a liquid layer containing recombinant interleukin-2 (IL-2), phytohemagglutinin (PHA), and B-cell growth factor over an agar feeder layer containing irradiated peripheral blood mononuclear cells plus fetal calf serum (FCS) and horse serum. Cultured cells expressed HLA-DR, CD-9, CD-10, CD-20, and CD-34 antigens, indicating conservation of precursor B phenotype. Differentiation antigen expression on the clonogenic subpopulation giving rise to leukemic colonies was assessed by treating cells prior to culture with selected cytotoxic monoclonal antibodies (MoAbs) and complement or by fluorescence-activated cell sorting. Lytic treatment with HLA-DR, CD- 10, CD-9, and CD-20 antibodies produced median reductions in colony formation of 93%, 81%, 73%, and 58% respectively. Combined treatment with CD-9 and CD-10 antibodies produced complete inhibition in 11 of 13 cases. Cell-sorting experiments after CD-10 staining indicated a good correlation with complement lysis studies and also demonstrated that the CD-19 antigen is expressed on a high proportion of clonogenic cells. Colony formation was also significantly reduced in 13 of 17 cases by lytic treatment of cells with the CD-33 antibody MY-9, suggesting expression of this “myeloid” lineage antigen on ALL clonogenic cells. The substantial case-to-case variation in expression of individual differentiation antigens indicates that leukemic cellular heterogeneity is a major consideration in ex vivo bone marrow purging using MoAbs and emphasizes the need for more critical evaluation of purging techniques.

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25

Hudson, AM, V. Makrynikola, A. Kabral und KF Bradstock. „Immunophenotypic analysis of clonogenic cells in acute lymphoblastic leukemia using an in vitro colony assay“. Blood 74, Nr. 6 (01.11.1989): 2112–20. http://dx.doi.org/10.1182/blood.v74.6.2112.bloodjournal7462112.

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A culture system was used to analyze the expression of membrane differentiation antigens on the proliferative or clonogenic fraction of cells from cases of common (precursor B) acute lymphoblastic leukemia (c-ALL). Colonies of leukemic cells were obtained in 18 of 20 cases after 1 week in culture in a liquid layer containing recombinant interleukin-2 (IL-2), phytohemagglutinin (PHA), and B-cell growth factor over an agar feeder layer containing irradiated peripheral blood mononuclear cells plus fetal calf serum (FCS) and horse serum. Cultured cells expressed HLA-DR, CD-9, CD-10, CD-20, and CD-34 antigens, indicating conservation of precursor B phenotype. Differentiation antigen expression on the clonogenic subpopulation giving rise to leukemic colonies was assessed by treating cells prior to culture with selected cytotoxic monoclonal antibodies (MoAbs) and complement or by fluorescence-activated cell sorting. Lytic treatment with HLA-DR, CD- 10, CD-9, and CD-20 antibodies produced median reductions in colony formation of 93%, 81%, 73%, and 58% respectively. Combined treatment with CD-9 and CD-10 antibodies produced complete inhibition in 11 of 13 cases. Cell-sorting experiments after CD-10 staining indicated a good correlation with complement lysis studies and also demonstrated that the CD-19 antigen is expressed on a high proportion of clonogenic cells. Colony formation was also significantly reduced in 13 of 17 cases by lytic treatment of cells with the CD-33 antibody MY-9, suggesting expression of this “myeloid” lineage antigen on ALL clonogenic cells. The substantial case-to-case variation in expression of individual differentiation antigens indicates that leukemic cellular heterogeneity is a major consideration in ex vivo bone marrow purging using MoAbs and emphasizes the need for more critical evaluation of purging techniques.

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26

Scott, Helge, Wohan Ek, Jacob Havnen, Helge Michalsen, Leif Brunvand, Hans Howlid und Per Brandtzaeg. „Serum Antibodies to Dietary Antigens“. Journal of Pediatric Gastroenterology and Nutrition 11, Nr. 2 (August 1990): 215–20. http://dx.doi.org/10.1002/j.1536-4801.1990.tb10090.x.

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Summary:We examined 1,541 consecutive serum samples from 707 children with suspected food intolerance and 32 with treated celiac disease (CD) for IgG and IgA antibody reactivities to antigens from gluten, egg, and cow's milk by an enzyme‐linked immunosorbent assay (ELISA). Samples from 72 patients showed increased IgA and/or IgG reactivity to gluten antigens; four were known CD patients not complying with a gluten‐free diet, 13 were suspected CD patients challenged with gluten, and 30 most likely had CD as suggested by small intestinal villous atrophy and histological and/or clinical improvement on a gluten‐free diet. The remainder with increased antigluten activity had other disorders that might have affected mucosal permeability. Nevertheless, the median IgA reactivity to gluten was significantly higher in the CD group, and the probability for CD increased from 25 to 100% when this reactivity was above 2.4 optical density (OD) units in our ELISA. Sixteen CD patients (but none of those without CD) had IgA reactivity to gluten higher than 2.4 OD units. We conclude that ELISA determinations of levels of serum antibodies reacting to dietary antigens is a valuable adjunct in the diagnosis of CD in children.

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27

Kishimoto, T., S. Goyert, H. Kikutani, D. Mason, M. Miyasaka, L. Moretta, T. Ohno et al. „Update of CD antigens, 1996.“ Journal of Immunology 158, Nr. 7 (01.04.1997): 3035–36. http://dx.doi.org/10.4049/jimmunol.158.7.3035.

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28

Clark, Edward A. „CD nomenclature for mouse antigens?“ Immunology Today 8, Nr. 6 (Januar 1987): 170. http://dx.doi.org/10.1016/0167-5699(87)90030-2.

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29

Qin, Lin, Jun Cao, Kun Shao, Fan Tong, Zhihang Yang, Ting Lei, Yazhen Wang et al. „A tumor-to-lymph procedure navigated versatile gel system for combinatorial therapy against tumor recurrence and metastasis“. Science Advances 6, Nr. 36 (September 2020): eabb3116. http://dx.doi.org/10.1126/sciadv.abb3116.

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Application of cancer vaccines is limited due to their systemic immunotoxicity and inability to satisfy all the steps, including loading of tumor antigens, draining of antigens to lymph nodes (LNs), internalization of antigens by dendritic cells (DCs), DC maturation, and cross-presentation of antigens for T cell activation. Here, we present a combinatorial therapy, based on a α-cyclodextrin (CD)–based gel system, DOX/ICG/CpG-P-ss-M/CD, fabricated by encapsulating doxorubicin (DOX) and the photothermal agent indocyanine green (ICG). Upon irradiation, the gel system exhibited heat-responsive release of DOX and vaccine-like nanoparticles, CpG-P-ss-M, along with chemotherapy- and phototherapy-generated abundant tumor-specific antigen storage in situ. The released CpG-P-ss-M acted as a carrier adsorbed and delivered antigens to LNs, promoting the uptake of antigens by DCs and DC maturation. Notably, combined with PD-L1 blocking, the therapy effectively inhibited primary tumor growth and induced tumor-specific immune response against tumor recurrence and metastasis.

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Santos, Fred Luciano Neves, Paola Alejandra Fiorani Celedon, Nilson Ivo Tonin Zanchin, Amanda Leitolis, Sandra Crestani, Leonardo Foti, Wayner Vieira de Souza, Yara de Miranda Gomes und Marco Aurélio Krieger. „Performance Assessment of a Trypanosoma cruzi Chimeric Antigen in Multiplex Liquid Microarray Assays“. Journal of Clinical Microbiology 55, Nr. 10 (19.07.2017): 2934–45. http://dx.doi.org/10.1128/jcm.00851-17.

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ABSTRACT Diagnosing chronic Chagas disease (CD) requires antibody–antigen detection methods, which are traditionally based on enzymatic assay techniques whose performance depend on the type and quality of antigen used. Previously, 4 recombinant chimeric proteins from the Instituto de Biologia Molecular do Paraná (IBMP-8.1 to 8.4) comprising immuno-dominant regions of diverse Trypanosoma cruzi antigens showed excellent diagnostic performance in enzyme-linked immunosorbent assays. Considering that next-generation platforms offer improved CD diagnostic accuracy with different T. cruzi -specific recombinant antigens, we assessed the performance of these chimeras in liquid microarrays (LMAs). The chimeric proteins were expressed in Escherichia coli and purified by chromatography. Sera from 653 chagasic and 680 healthy individuals were used to assess the performance of these chimeras in detecting specific anti- T. cruzi antibodies. Accuracies ranged from 98.1 to 99.3%, and diagnostic odds ratio values were 3,548 for IBMP-8.3, 4,826 for IBMP-8.1, 7,882 for IBMP-8.2, and 25,000 for IBMP-8.4. A separate sera bank (851 samples) was employed to assess cross-reactivity with other tropical diseases. Leishmania , a pathogen with high similarity to T. cruzi , showed cross-reactivity rates ranging from 0 to 2.17%. Inconclusive results were negligible (0 to 0.71%). Bland–Altman and Deming regression analysis based on 200 randomly selected CD-positive and negative samples demonstrated interchangeability with respect to CD diagnostic performance in both singleplex and multiplex assays. Our results suggested that these chimeras can potentially replace antigens currently used in commercially available assay kits. Moreover, the use of multiplex platforms, such as LMA assays employing 2 or more IBMP antigens, would abrogate the need for 2 different testing techniques when diagnosing CD.

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Favaloro, Emmanuel J., Nick Moraitis, Jerry Koutts, Thomas Exner und Kenneth F. Bradstock. „Endothelial Cells and Normal Circulating Haemopoietic Cells Share a Number of Surface Antigens“. Thrombosis and Haemostasis 61, Nr. 02 (1989): 217–24. http://dx.doi.org/10.1055/s-0038-1646562.

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SummaryHuman endothelial cells, cultured from umbilical cord veins, have been evaluated for expression of a large number of cell surface antigens with known haemopoietic, particularly myeloid, cell distribution. This was achieved by evaluating endothelial reactivity (using non-fixed cells) with groups of monoclonal antibodies (MAB) belonging to distinct Clusters of Differentiation (CD), as defined by the Third International Workshop on Leukocyte Differentiation Antigens (ILWS). Results indicate that many antigens known to be present on haemopoietic cells, including those on platelets, are co-expressed on endothelial cells. The most intense degree of reactivity was seen using MAB to CD-9 and CD-13, although significant reactivity was also observed using MAB to CD-31 and CD-32. Data also suggests weak binding to endothelial cells of MAB belonging to CD-14, CD-15 and CD-16. A number of unclustered MAB reactive with haemopoietic antigens can also be shown to bind to endothelial cells.

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Furue, Masutaka, und Yasumasa Ishibashi. „Differential localization of CD]a, CD]b, CD]c antigens in normal human skin“. Journal of Dermatological Science 2, Nr. 3 (Mai 1991): 241. http://dx.doi.org/10.1016/0923-1811(91)90198-7.

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Escribano, Luis, Beatriz Díaz-Agustín, Rosa Núñez, Aranzazu Prados, Ramón Rodríguez und Alberto Orfao. „Abnormal Expression of CD Antigens in Mastocytosis“. International Archives of Allergy and Immunology 127, Nr. 2 (2002): 127–32. http://dx.doi.org/10.1159/000048183.

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Razim, Agnieszka, Sabina Górska und Andrzej Gamian. „Non-Toxin-Based Clostridioides difficile Vaccination Approaches“. Pathogens 12, Nr. 2 (02.02.2023): 235. http://dx.doi.org/10.3390/pathogens12020235.

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Clostridioides difficile (CD) is a Gram-positive, anaerobic bacterium that infects mainly hospitalized and elderly people who have been treated with long-term antibiotic therapy leading to dysbiosis. The deteriorating demographic structure and the increase in the number of antibiotics used indicate that the problem of CD infections (CDI) will continue to increase. Thus far, there is no vaccine against CD on the market. Unfortunately, clinical trials conducted using the CD toxin-based antigens did not show sufficiently high efficacy, because they did not prevent colonization and transmission between patients. It seems that the vaccine should also include antigens found in the bacterium itself or its spores in order not only to fight the effects of toxins but also to prevent the colonization of the patient. This literature review summarizes the latest advances in research into vaccine antigens that do not contain CD toxins.

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Santos, Emily F., Ramona T. Daltro, Carlos G. Regis-Silva, Tycha B. S. Pavan, Fabrícia A. de Oliveira, Ângela M. da Silva, Roque P. Almeida et al. „Assessment of Cross-Reactivity of Chimeric Trypanosoma cruzi Antigens with Crithidia sp. LVH-60A: Implications for Accurate Diagnostics“. Diagnostics 13, Nr. 22 (17.11.2023): 3470. http://dx.doi.org/10.3390/diagnostics13223470.

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This study focuses on developing accurate immunoassays for diagnosing Chagas disease (CD), a challenging task due to antigenic similarities between Trypanosoma cruzi and other parasites, leading to cross-reactivity. To address this challenge, chimeric recombinant T. cruzi antigens (IBMP-8.1, IBMP-8.2, IBMP-8.3, and IBMP-8.4) were synthesized to enhance specificity and reduce cross-reactivity in tests. While these antigens showed minimal cross-reactivity with leishmaniasis, their performance with other trypanosomatid infections was unclear. This study aimed to assess the diagnostic potential of these IBMP antigens for detecting CD in patients with Crithidia sp. LVH-60A, a parasite linked to visceral leishmaniasis-like symptoms in Brazil. This study involved seven Crithidia sp. LVH-60A patients and three Leishmania infantum patients. The results indicated that these IBMP antigens displayed 100% sensitivity, with specificity ranging from 87.5% to 100%, and accuracy values between 90% and 100%. No cross-reactivity was observed with Crithidia sp. LVH-60A, and only one L. infantum-positive sample showed limited cross-reactivity with IBMP-8.1. This study suggests that IBMP antigens offer promising diagnostic performance, with minimal cross-reactivity in regions where T. cruzi and other trypanosomatids are prevalent. However, further research with a larger number of Crithidia sp. LVH-60A-positive samples is needed to comprehensively evaluate antigen cross-reactivity.

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Hundorfean, Gheorghe, Klaus-Peter Zimmer, Stephan Strobel, Andreas Gebert, Diether Ludwig und Jürgen Büning. „Luminal antigens access late endosomes of intestinal epithelial cells enriched in MHC I and MHC II molecules: in vivo study in Crohn's ileitis“. American Journal of Physiology-Gastrointestinal and Liver Physiology 293, Nr. 4 (Oktober 2007): G798—G808. http://dx.doi.org/10.1152/ajpgi.00135.2007.

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In contrast to healthy conditions, intestinal epithelial cells (IECs) stimulate proinflammatory CD4+and CD8+T cells during Crohn's disease (CD). The underlying regulatory mechanisms remain unknown. Here we investigated the epithelial expression of major histocompatibility complex (MHC) I and MHC II and its interference with endocytic pathways, in vivo. During ileoscopy, ovalbumin (OVA) was sprayed onto ileal mucosa of CD patients (ileitis and remission) and controls. The epithelial traffic of OVA and MHC I/II pathways were studied in biopsies using fluorescence and electron microscopy. We found MHC I and MHC II to accumulate within multivesicular late endosomes (MVLE) of IECs. Faint labeling for these molecules was seen in early endosomes and lysosomes. MVLE were entered by OVA 10 min after exposure. Exosomes carrying MHC I, MHC II, and OVA were detected in intercellular spaces of the epithelium. OVA trafficking and labeling patterns for MHC I and MHC II in IECs showed no differences between CD patients and controls. Independent of inflammatory stimuli, MHC I and MHC II pathways intersect MVLE in IECs, which were efficiently targeted by luminal antigens. Similar to MHC II-enriched compartments in professional antigen presenting cells, these MVLE might be critically involved in MHC I- and MHC II-related antigen processing in IECs and the source of epithelial-released exosomes. The access of luminal antigens to MHC I in MVLE might indicate that the presentation of exogenous antigens by IECs must not be restricted to MHC II but might also occur as “cross-presentation” via MHC I to CD8+T cells.

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Zola, Heddy. „Human leukocyte differentiation antigens as therapeutic targets: the CD molecules and CD antibodies“. Expert Opinion on Biological Therapy 1, Nr. 3 (Mai 2001): 375–83. http://dx.doi.org/10.1517/14712598.1.3.375.

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Kumar, S., T. Lutteke und R. Schwartz-Albiez. „GlycoCD: a repository for carbohydrate-related CD antigens“. Bioinformatics 28, Nr. 19 (30.07.2012): 2553–55. http://dx.doi.org/10.1093/bioinformatics/bts481.

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Daltro, Ramona Tavares, Emily Ferreira Santos, Ângelo Antônio Oliveira Silva, Natália Erdens Maron Freitas, Leonardo Maia Leony, Larissa Carvalho Medrado Vasconcelos, Alejandro Ostermayer Luquetti et al. „Western blot using Trypanosoma cruzi chimeric recombinant proteins for the serodiagnosis of chronic Chagas disease: A proof-of-concept study“. PLOS Neglected Tropical Diseases 16, Nr. 11 (28.11.2022): e0010944. http://dx.doi.org/10.1371/journal.pntd.0010944.

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Background Chagas disease (CD) is caused by Trypanosoma cruzi. The chronic phase of CD is characterized by the presence of IgG anti-T. cruzi antibodies; and diagnosis is performed by serological methods. Because there is no realible test that can be used as a reference test, WHO recommends the parallel use of two different tests for CD serodiagnosis. If results are inconclusive, samples should be subjected to a confirmatory test, e.g., Western blot (WB) or PCR. PCR offers low sensitivity in the chronic phase, whereas few confirmatory tests based on the WB method are commercially available worldwide. Therefore, new diagnostic tools should be evaluated to fill the gap in confirmatory tests CD. In recent years, four chimeric recombinant antigens (IBMP-8.1, IBMP-8.2, IBMP-8.3 and IBMP-8.4) have been evaluated in phase I, II and III studies using ELISA, liquid microarray and immunochromatography with 95–100% accuracy. Given the high diagnostic performance of these antigens, the present study investigated the ability of these molecules to diagnose chronic CD using a WB testing platform. Methodology/Principal findings In this study, we analyzed the diagnostic potential of four chimeric antigens using 40 T. cruzi-positive, 24-negative, and three additional samples positive for visceral leishmaniasis (i.e., potentially cross-reactive) using WB as the diagnostic platform. Checkerboard titration with different dilutions of antigens, conjugated antigens, and serum samples was performed to standardize all assays. All IBMP antigens achieved 100% sensitivity, specificity, and accuracy, with the exception of IBMP-8.3, which had 100% specificity despite lack of significance, but lower sensitivity (95%) and accuracy (96.9%). No cross-reactivity was observed in samples positive for leishmaniasis. Conclusions/Significance The present phase I (proof-of-concept) study demonstrated the high diagnostic potential of these four IBMP antigens to discriminate between T. cruzi-positive and -negative samples, making them candidates for phase II and confirmatory testing with WB.

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Shaw, Stephen. „Leukocyte differentiation antigens how to keep up with the litany of CD antigens“. Human Immunology 41, Nr. 2 (Oktober 1994): 103–4. http://dx.doi.org/10.1016/0198-8859(94)90001-9.

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Chen, Huan-Yuan, Jun Yang, Ahmet Bora Inceoglu, Li-Chieh Wang, Lan Yu, Bruce Hammock und Fu-Tong Liu. „Identification and analysis of lipid mediators in a mouse model of allergic contact dermatitis (163.9)“. Journal of Immunology 186, Nr. 1_Supplement (01.04.2011): 163.9. http://dx.doi.org/10.4049/jimmunol.186.supp.163.9.

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Abstract Allergic contact dermatitis (CD) is a type of inflammatory skin disease resulting from delayed-typed hypersensitivity to haptenic antigens. The antigens are presented to T cells by dendritic cells in the skin and result in activation and recruitment of other immune cells to the skin. The inflammatory response is frequently accompanied by production of metabolic mediators. Here we sought to identify mediators involved in allergic CD by using a mouse model. C57BL/6 mice were sensitized with dinitrofluorobenzene or oxazolone and challenged with the same antigen on the ear. The treated ears were swollen and exhibited accumulation of leukocytes. The affected ears were subjected to targeted quantitative metabolomic analysis by mass spectrometry to identify lipid metabolites that displayed significant changes. A cyclooxygenase product, thromboxane A2, was remarkably induced by more than 10 fold compared to vehicle control. A number of other bioactive lipid metabolites were induced by more than two fold. We found that mice treated with aspirin or sEHi showed reduced ear thickness compared to vehicle treatment. Our data suggest that the cyclooxygenase and soluble epoxide hydrolase pathways are involved in the development of allergic CD by modulating the inflammatory skin response. Inhibitors of these two pathways and antagonists of prostanoid receptors for the elevated metabolites are expected to be effective therapeutically to treat allergic CD.

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Lai, Lily, Noosheen Alaverdi, Lois Maltais und Herbert C. Morse. „Mouse Cell Surface Antigens: Nomenclature and Immunophenotyping“. Journal of Immunology 160, Nr. 8 (15.04.1998): 3861–68. http://dx.doi.org/10.4049/jimmunol.160.8.3861.

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Abstract This paper reviews cell surface Ags expressed on mouse hemopoietic and nonhemopoietic cells. The review will cover molecules included in the cluster of differentiation (CD) from CD1 to CD166 and lymphocyte Ag (Ly) series from Ly-1 to Ly-81 as well as some new Ags without current CD or Ly assignments. In addition to an update on mouse nomenclature, there will be a discussion of some known functions of the molecules and brief comments on the use of particular Ags for immunophenotyping of cell subsets. Several novel markers mentioned may prove useful in mouse immunology research.

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Bonamico, M., P. Mariani, M. C. Mazzilli, P. Triglione, P. Lionetti, P. Ferrante, A. Picarelli, A. Mesturino, G. Gemme und C. Imperato. „Frequency and Clinical Pattern of Celiac Disease Among Siblings of Celiac Children“. Journal of Pediatric Gastroenterology and Nutrition 23, Nr. 2 (August 1996): 159–63. http://dx.doi.org/10.1002/j.1536-4801.1996.tb00321.x.

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SummaryTo investigate the prevalence and clinical and genetic patterns of celiac disease (CD) among siblings of CD patients, 103 siblings and one twin of 80 celiac children were evaluated by means of their clinical history, physical examination, blood indices of nutritional status, and antigliadin antibodies (AGA). Antiendomysium antibody (AEA) levels were determined in 70 patients and 85 subjects were human leucocyte antigen (HLA) typed. On the basis of clinical or laboratory data or both, 21 siblings (20.2%) were submitted to intestinal biopsy, whereas intestinal biopsy in six siblings with positive serologic screening (AGA lgA or AEA or both) was not performed because of parental refusal. In a high percentage of cases (18%), all on a gluten‐containing diet, the intestinal mucosa was atrophic, and CD was subsequently diagnosed. Because we could not submit all the siblings to intestinal biopsy, this figure could underestimate the real prevalence of the disease in our series; consequently, it was not possible to calculate accurately the sensitivity and specificity of AGA and AEA. Nevertheless, AEA (positive in all the nine siblings with mucosal atrophy), followed by AGA IgA, proved to be the best screening for CD. Eighteen of 19 CD siblings showed HLA‐predisposing antigens. Among the 19 CD siblings, one showed a typical form with gastrointestinal symptoms, two had short stature, one suffered from recurrent vomiting, and in 15, the disease was clinically silent. On the contrary, among siblings who were first diagnosed (index cases), the majority (73.7%) had a typical from of CD, and no clinically silent cases were observed. We did not find any difference between index cases and CD siblings in food habits and distribution of HLA antigens. In 15 of 18 cases, the sibling diagnosed subsequently was the older one. Finally, the typical form of CD was significantly more frequent among the younger brother than the older. In conclusion, the high prevalence of the silent form of CD in our cases indicates that siblings of CD subjects should always be screened for CD. The combination of AGA IgA and AEA represent a good screening method to use in selecting children for the intestinal biopsy.

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Vojvodic, Svetlana, und Dusica Ademovic-Sazdanic. „HLA II class antigens and susceptibility to coeliac disease“. Genetika 43, Nr. 3 (2011): 517–26. http://dx.doi.org/10.2298/gensr1103517v.

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Coeliac disease (CD) is a systemic autoimmune, complex and multifactorial disorder, which is caused by interactions between genetic and environmental factors. The only established genetic risk factors so far are the human leucocyte antigens. The aim of this study was to assess the distribution of II class human leukocyte antigens (HLA) in patients with coeliac disease and to investigate the susceptibility to coeliac disease in family members. We typed HLA DR and DQ antigens in 37 patients from Vojvodina with coeliac disease, 23 first-degree relatives, and 210 controls, serologically using standard lymphocytotoxicity technique. HLA DQ5(1), DQ6(1), DR11(5), DQ7(3), DQ2 and DR15(2) were the most common antigens in the control group. Frequency of HLA DQ2, DR3 and DR7 was higher in CD patients than in the control group. The relative risks for HLA DQ2, DR3 and DR7 were 4.846, 6.986 and 2.106, respectively, while positive association was found between HLA DQ2 and DR3 and CD. Frequency of HLA DQ2, DR3 and DR16(2) was higher in first-degree relatives than in the control group while a positive association was found between HLA DQ2 and DR3. A negative association was found between HLA DQ5(1) and DQ6(1) in coeliac patients from Vojvodina and their relatives, in addition to HLA DR11(5) in the group of relatives (RR=0.363,PF=0.232). These findings indicate the impact of the HLA testing for CD in clinical practice in order to rule out the possibility to CD in doubtful cases or in at-risk subjects.

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Celedon, Paola Alejandra Fiorani, Leonardo Maia Leony, Ueriton Dias Oliveira, Natália Erdens Maron Freitas, Ângelo Antônio Oliveira Silva, Ramona Tavares Daltro, Emily Ferreira Santos, Marco Aurélio Krieger, Nilson Ivo Tonin Zanchin und Fred Luciano Neves Santos. „Stability Assessment of Four Chimeric Proteins for Human Chagas Disease Immunodiagnosis“. Biosensors 11, Nr. 8 (22.08.2021): 289. http://dx.doi.org/10.3390/bios11080289.

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The performance of an immunoassay relies on antigen-antibody interaction; hence, antigen chemical stability and structural integrity are paramount for an efficient assay. We conducted a functional, thermostability and long-term stability analysis of different chimeric antigens (IBMP), in order to assess effects of adverse conditions on four antigens employed in ELISA to diagnose Chagas disease. ELISA-based immunoassays have served as a model for biosensors development, as both assess molecular interactions. To evaluate thermostability, samples were heated and cooled to verify heat-induced denaturation reversibility. In relation to storage stability, the antigens were analyzed at 25 °C at different moments. Long-term stability tests were performed using eight sets of microplates sensitized. Antigens were structurally analyzed through circular dichroism (CD), dynamic light scattering, SDS-PAGE, and functionally evaluated by ELISA. Data suggest that IBMP antigens are stable, over adverse conditions and for over a year. Daily analysis revealed minor changes in the molecular structure. Functionally, IBMP-8.2 and IBMP-8.3 antigens showed reactivity towards anti-T. cruzi antibodies, even after 72 h at 25 °C. Long-term stability tests showed that all antigens were comparable to the control group and all antigens demonstrated stability for one year. Data suggest that the antigens maintained their function and structural characteristics even in adverse conditions, making them a sturdy and reliable candidate to be employed in future in vitro diagnostic tests applicable to different models of POC devices, such as modern biosensors in development.

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Becker, Pablo D., Gustavo M. Bertot, David Souss, Thomas Ebensen, Carlos A. Guzmán und Saúl Grinstein. „Intranasal Vaccination with Recombinant Outer Membrane Protein CD and Adamantylamide Dipeptide as the Mucosal Adjuvant Enhances Pulmonary Clearance of Moraxella catarrhalis in an Experimental Murine Model“. Infection and Immunity 75, Nr. 4 (13.11.2006): 1778–84. http://dx.doi.org/10.1128/iai.01081-06.

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ABSTRACT Moraxella catarrhalis causes acute otitis media in children and lower respiratory tract infections in adults and elderly. In children the presence of antibodies against the highly conserved outer membrane protein CD correlates with protection against infection, suggesting that this protein may be useful as a vaccine antigen. However, native CD is difficult to purify, and it is still unclear if recombinant CD (rCD) is a valid alternative. We performed a side-by-side comparison of the immunogenicities and efficacies of vaccine formulations containing native CD and rCD with adamantylamide dipeptide as the mucosal adjuvant. Intranasal vaccination of mice stimulated the production of high CD-specific antibody titers in sera and of secretory immunoglobulin A in mucosal lavages, which cross-recognized both antigens. While vaccination with native CD increased the number of interleukin-2 (IL-2)- and gamma interferon-producing cells, rCD mainly stimulated IL-4-secreting cells. Nevertheless, efficient bacterial clearance was observed in the lungs of challenged mice receiving native CD and in the lungs of challenged mice receiving rCD (96% and 99%, respectively). Thus, rCD is a promising candidate for incorporation in vaccine formulations for use against M. catarrhalis.

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Srinivasan, Bharani, Margit Focke-Tejkl, Ines Swoboda, Claudia Constantin, Irene Mittermann, Sandra Pahr, Harald Vogelsang, Wolf-Dietrich Huber und Rudolf Valenta. „Molecular characterization of wheat antigens involved in celiac disease (71.13)“. Journal of Immunology 188, Nr. 1_Supplement (01.05.2012): 71.13. http://dx.doi.org/10.4049/jimmunol.188.supp.71.13.

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Abstract Wheat gliadins trigger a hypersensitive immune response in genetically susceptible individuals causing celiac disease (CD). The small intestinal mucosa of CD patients is characterised by presence of numerous infiltrating CD4+ T cells. These T cells proliferate and secrete IFN-γ when stimulated with gliadin extracts and are implicated in the mucosal damage and production of anti-gliadin antibodies. Peptide sequences in the Pro(P) and Glub(Q) rich regions of gliadins are known to be T cell-stimulatory but due to the heterogeneity of gliadin sequences, several potential stimulatory epitopes and the individual protein antigens are yet to be identified and characterized. Thus our aim was to identify and characterize wheat antigens with ability to initiate and sustain celiac disease. Due to their complex biochemistry the three subtypes of gliadins α/β, γ, and ω-gliadins are difficult to isolate into pure fractions. Hence, we developed a method wherein the alcohol extracted gliadins was sub-fractionated ion-exchange chromatography and each sub-fraction’s reactivity to serum IgA, from clinically well defined CD (active/diet) patients and non-CD patients was analyzed. We identified and generated recombinant gliadins that were CD specific. Epitope mapping studies revealed major immune-reactive regions. Recombinant gamma-gliadins will be useful for characterizing the immune response to wheat antigens and to develop reliable diagnostic and therapeutic strategies.

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Brandt, Sven, Krystin Krauel, Kay E. Gottschalk, Thomas Renné, Christiane A. Helm, Andreas Greinacher und Stephan Block. „Characterisation of the conformational changes in platelet factor 4 induced by polyanions: towards in vitro prediction of antigenicity“. Thrombosis and Haemostasis 112, Nr. 07 (2014): 53–64. http://dx.doi.org/10.1160/th13-08-0634.

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SummaryHeparin-induced thrombocytopenia (HIT) is the most frequent drug-induced immune reaction affecting blood cells. Its antigen is formed when the chemokine platelet factor 4 (PF4) complexes with polyanions. By assessing polyanions of varying length and degree of sulfation using immunoassay and circular dichroism (CD)-spectroscopy, we show that PF4 structural changes resulting in antiparallel β-sheet content >30% make PF4/polyanion complexes antigenic. Further, we found that polyphosphates (polyP-55) induce antigenic changes on PF4, whereas fondaparinux does not. We provide a model suggesting that conformational changes exposing antigens on PF4/polyanion complexes occur in the hairpin involving AA 32–38, which form together with C-terminal AA (66–70) of the adjacent PF4 monomer a continuous patch on the PF4 tetramer surface, explaining why only tetrameric PF4 molecules express “HIT antigens”. The correlation of antibody binding in immunoassays with PF4 structural changes provides the intriguing possibility that CD-spectroscopy could become the first antibody-independent, in vitro method to predict potential immunogenicity of drugs. CD-spectroscopy could identify compounds during preclinical drug development that induce PF4 structural changes correlated with antigenicity. The clinical relevance can then be specifically addressed during clinical trials. Whether these findings can be transferred to other endogenous proteins requires further studies.

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Singer, JW, P. Charbord, A. Keating, J. Nemunaitis, G. Raugi, TN Wight, JA Lopez, GJ Roth, LW Dow und PJ Fialkow. „Simian virus 40-transformed adherent cells from human long-term marrow cultures: cloned cell lines produce cells with stromal and hematopoietic characteristics“. Blood 70, Nr. 2 (01.08.1987): 464–74. http://dx.doi.org/10.1182/blood.v70.2.464.464.

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Abstract Adherent cells from long-term marrow cultures from 23 individuals were transformed with wild-type simian virus 40 (SV40). After transformation, cloned cell lines were developed that even after rigorous subcloning invariably produced both stromal cells and round cells. The stromal cells expressed cytoskeletal filaments similar to those of long-term marrow culture adherent cells and produced interstitial and basal lamina collagen types. The round cells had the electron microscopic appearance of primitive hematopoietic cells and when examined with cytochemical stains and monoclonal antibodies to hematopoietic differentiation antigens had reaction patterns suggestive of cells from several lineages. Most round cells expressed the pan- hematopoietic T-200 determinant, and lesser percentages expressed the early T cell antigens CD-1 and CD-3, HLA-DR determinants, the monocytic antigen recognized by Leu M3, and the myeloid antigens detected by monoclonal antibodies 1G10 and 12.8. In addition, when plated in semisolid medium in the presence of a source of colony-stimulating activity, up to 11% of the cells formed colonies consisting of blastlike cells that also expressed hematopoietic cell surface determinants. The data suggest that adherent cells in long-term marrow cultures contain a cell that after transformation by SV40 obligately produces cells with hematopoietic as well as stromalike features.

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Singer, JW, P. Charbord, A. Keating, J. Nemunaitis, G. Raugi, TN Wight, JA Lopez, GJ Roth, LW Dow und PJ Fialkow. „Simian virus 40-transformed adherent cells from human long-term marrow cultures: cloned cell lines produce cells with stromal and hematopoietic characteristics“. Blood 70, Nr. 2 (01.08.1987): 464–74. http://dx.doi.org/10.1182/blood.v70.2.464.bloodjournal702464.

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Adherent cells from long-term marrow cultures from 23 individuals were transformed with wild-type simian virus 40 (SV40). After transformation, cloned cell lines were developed that even after rigorous subcloning invariably produced both stromal cells and round cells. The stromal cells expressed cytoskeletal filaments similar to those of long-term marrow culture adherent cells and produced interstitial and basal lamina collagen types. The round cells had the electron microscopic appearance of primitive hematopoietic cells and when examined with cytochemical stains and monoclonal antibodies to hematopoietic differentiation antigens had reaction patterns suggestive of cells from several lineages. Most round cells expressed the pan- hematopoietic T-200 determinant, and lesser percentages expressed the early T cell antigens CD-1 and CD-3, HLA-DR determinants, the monocytic antigen recognized by Leu M3, and the myeloid antigens detected by monoclonal antibodies 1G10 and 12.8. In addition, when plated in semisolid medium in the presence of a source of colony-stimulating activity, up to 11% of the cells formed colonies consisting of blastlike cells that also expressed hematopoietic cell surface determinants. The data suggest that adherent cells in long-term marrow cultures contain a cell that after transformation by SV40 obligately produces cells with hematopoietic as well as stromalike features.

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