Production and Characterization of Human Monoclonal Antibodies against the Organophosphorus Poison Soman

摘要

The technical objective of this project is to produce human anti-soman antibodies from human lymphocytes in vitro and to develop such antibodies as monoclonal products with high affinity for Soman. During the period covered by this annual report (October 1, 1983 - September 30, 1984) we have: Prepared conjugates of Soman with various immunogenic carriers; Developed anti-Soman plaque-forming cell (PFC) assays and an ELISA to detect anti-Soman antibodies and used these assays to evaluate the immunogenicity of Soman conjugates; Developed an in vitro tissue culture system for human tonsillar lymphocytes that incorporates B cell inducing factors in order to permit near optimal development of antibody-producing cells; Produced murine hybridomas representing different immunoglobulin isotypes for use as standards for comparison with human anti-Soman hybridoma antibodies; Established a method for transforming human fibroblast cell lines in vitro allowing their immortalization; Constructed a hybrid tumor virus from SV40 and a xenotropic murine leukemia virus which has transformed 17 human cell lines; Developed conditions that allow cocultivation of human tonsil cells with the xenotropic hybrid transforming virus as a result of which viral T antigen expression could be demonstrated in the lymphocytes; and acquired a library of other oncogenes that have been demonstrated to immortalize rodent cells in culture thus paving the way for using oncogene transfection as an alternative to viral transformation in order to achieve B cell immortalization.