Preparation of human monoclonal antibodies using human B cell producing autoantibodies and analyses of novel autoantigens.

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In some cases with autoimmune diseases and malignancies, autoantibodies to unknown antigens were detected in the sera. Using monoclonal antibodies, we have studied these autoantigens and autoantibodies and analysed the immune system. In order to prepare human type monoclonal antibodies, human B cells from cases with autoimmune diseases and malignancies were used. Therein we experienced a colonic cancer patient with anti-CEA monoclonal antibody. For parent cells, human myeloma cell line ARH77, human plasma cell line UC729-6 and mouse myeloma cell line P3-X63-Ag8-653 were served. Human cell lines which we used for fusion gave little hybridomas, but many human monoclonal antibodies were obtained when P3-X63-Ag8-653 was used. By a fusion with B cells in pleural effusion of a gastric cancer patient, human monoclonal antibody 3B7 (IgG) was raised, showing a staining reactivity with gastric and colonic cancers but not with non cancerous tissues. Molecular profile of autoantigen to which monoclonal antibody 3B7 reacts is under a further investigation. At present we are trying to raise hybridomas from new parent cells KR-4 and KR-12 using a newly developed electrofusion method.