Inhibition of murine T cell-mediated cytolysis and T cell proliferation by a rat monoclonal antibody immunoprecipitating two lymphoid cell surface polypeptides of 94 000 and 180 000 molecular weight

Abstract

The monoclonal antibody methodology was use to identify membrane structures involved in T cell functions. To optimize chances to produce and detect relevant antibodies, a xenogeneic sensitization protocol was utilized and hybridoma supernatants were screened, on functional rather than structural grounds, for their ability to inhibit a given function. The test function was T cell-mediated cytolysis. Mouse cytolytic anti-allogeneic cell populations were used to sensitize a rat, the spleen cells of which were fused to produce hybridomas; the supernatants of the latter were screened for their ability to inhibit mouse T cell-mediated cytolysis in vitro. Several inhibitory antibodies were obtained, one of which, H35-89.9 monoclonal antibody, was studied in more detail. It inhibited specific and concanavalin A (Con A)-mediated cytolysis by T cells, by acting on the effector cells. It reversibly inhibited soluble antigen-, alloantigen and Con A-induced T cell proliferation (but not LPS-induced B cell proliferation), after the production of interleukin 2, by acting on the responder cells. It also had a desagglutinating effect on Con A and LPS blasts and on EL4 cells. In immunoprecipitated from thymocyte membrane preparations two structures of 94 000 and 180 000 apparent molecular weight, and recognized cell surface determinants on both T and B lymphocytes. Our findings suggest that several antibodies directed against distinct effector cell membrane structures inhibit cytolysis. The case of H35-89.9 monoclonal antibody, which exerts multiple functional effects and immunoprecipitates two membrane polypeptides, raises the problem of the various possible relationships between these structures and functions.