Making the antibody library from pathologic lesion of intestinal bowel disease, and analysis and application of antibody and antigen.

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The purpose of our research is to find useful antibodies and antigens for analyzing the pathogenesis and for diagnosing and treating inflammatory bowel diseases (IBDs). In the lesions of IBDs, numbers of antibody-producing plasma cells infiltrate. However, the antigens recognized by antibodies produced in these immune cells remain unclear. Theoretically, the antibodies locally secreted within the lesions must be closely correlated with the disease process. We thus planned a research directly analyzing the antibodies produced by the plasma cells infiltrating in the lesion.In the first year, we examined the clonality of cDNA encoding antibodies in mouse inflammatory colonic lesions experimentally provoked by dextran sodium sulfate administration, which represents a mouse model of ulcerative colitis. We identified clonalities of the antibody-producing cells infiltrating within the colonic lesion. Therefore, we assumed that the clonality of cDNA encoding antibodies might be useful as crite … More ria for identifying lesion-specific antibodies.In the second year, we analyzed the clonality of antibody cDNA in regional lymph nodes obtained from horseradish peroxidase (HRP)-immunized rats. The HRP reactivity of single chain Fv (scFv) monoclonal antibodies synthesized from nodal tissue cDNA was pursued. We demonstrated clonalities in the nodal tissue cDNA encoding both the heavy and light chains. However, scFv monoclonal antibodies randomly synthesized from the cDNA lacked binding with HRP in the ELISA system. In order to identify cDNA encoding anti-HRP antibodies in the lymph nodes, we screened the cDNA library made from the immunized nodal tissue. Only one type of antibody clone was observed. By random sequence analysis, the heavy chain cDNA of this anti-HRP monoclonal antibody corresponded to one of the several clones. However, the light chain cDNA of the anti-HRP antibody was identical to the sequence that did not show any clonality. These results suggest to our regret that cDNA clones encoding antibody molecules do not necessarily indicate the lesion-specific antibodies. To overcome this difficulty, further analysis is now continuing. Less