Production of monoclonal antibodies reacting with peripheral blood mononuclear cell surface differentiation antigens.

Abstract

The present communication describes the generation of 22 lymphocyte hybrid cell lines whose antibodies react with the human HSB-2 T cell line, and the initial characterization of the antibodies synthesized by five of these cell lines. Three of the lymphocyte hybridomas produced antibodies that specifically bound to the cell surface of HSB-2 T cells but did not bind to cells of the autologous B cell line, designated SB. With these three antibodies, 10,000 times the concentration of antibody necessary to produce detectable binding of antibody to HSB-2 cells failed to produce detectable binding to SB cells. Of the three HSB-2-specific antibodies that were characterized by using fluorescent labeling techniques, one did not bind to any peripheral blood (PB) leukocytes (5A12), one bound to small numbers of both PB T and complement receptor-positive (CR+) lymphocytes (7E6), and one antibody bound specifically only to a subset of PB T lymphocytes (3A1). Of two antibodies that bound to both HSB-2 and SB cells, by fluorescent labeling, one (3F10) bound to all PB leukocytes whereas the other (4F2) bound primarily to PB monocytes and to a lesser extent to PB T cells. This study illustrates the feasibility and simplicity of generating cell lines producing monoclonal antibodies to lymphocyte cell surface differentiation antigens and describes several monoclonal antibodies that should facilitate studies of human PB mononuclear cells.