Preparation,characterization and application of anti-human OX40 ligand (OX40L) monoclonal antibodies and establishment of a sandwich ELISA for autoimmune diseases detection |
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| Institution: | 1. Jiangsu Institute of Clinical Immunology, The First Affiliated Hospital of Soochow University, Suzhou, China;2. Jiangsu Key Laboratory of Clinical Immunology, The First Affiliated Hospital of Soochow University, Suzhou, China;3. Institute of Pediatrics, Children''s Hospital of Soochow University, Suzhou, China;4. Institute of Blood and Marrow Transplantation, Soochow University, Suzhou, China;1. Department of Immunology and Rheumatology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan;2. Center for Bioinformatics and Molecular Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan;3. Department of Rheumatology, Clinical Research Center, National Hospital Organization Nagasaki Medical Center;4. Department of Rheumatology, Sasebo Chuo Hospital, Nagasaki, Japan;5. Biomedical Research Support Center, Nagasaki University School of Medicine, Nagasaki, Japan;6. Department of Endocrinology and Metabolism, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan;7. Department of Rehabilitation Sciences, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan;1. Department of Urology, Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, China;2. Department of Pathology, Ningbo Diagnostic Pathology Center, Ningbo, Zhejiang 315000, China;1. The Laboratory of Lipid & Glucose Metabolism, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China;2. Center for Cognitive and Neurobiological Imaging, Stanford University, Stanford, CA, USA;3. Department of Emergency and Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China;1. Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, China;2. School of Life Science, Jilin University, Changchun, China;3. China-Japan Union Hospital of Jilin University, Changchun, China |
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| Abstract: | OX40L (CD252, TNFSF4), a type II transmembrane protein which like other tumor necrosis factor ligands, involved in the costimulation and differentiation of T cells, functions as a positive signal in immune response. To investigate the biological function of soluble OX40L (sOX40L), three functional anti-OX40L monoclonal antibodies (mAbs) 3D2, 3F7 and 2H3 were obtained by hybridoma technology. Besides, specificity of the mAbs was further demonstrated by ELISA, Western blot and Immunofluorescence experiments. We also developed a novel enzyme-linked immunosorbent assay (ELISA) based on two anti-human OX40L antibodies 3D2 and 3F7 with different epitopes. Using the ELISA system, we found that sOX40L in the sera of healthy donors increases in an age-dependent manner and that enhanced sOX40L expression in some autoimmune diseases especially in rheumatoid arthritis (RA) patients, suggesting the potential diagnostic significance of sOX40L in the autoimmune diseases. Together, these data demonstrate that the existence of circulating sOX40L in human sera might play an important role in immunoregulation. |
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