Comparative Analysis of Natural Antibody Specificities among Hybridomas Originating from Spleen and Peritoneal Cavity of Adult NZB and BALB/c Mice

Abstract

A preliminary experiment showed that the supernatants of in vitro cultured peritoneal cells (rich in Ly‐1 B cell subset shown to secrete most IgM autoantibodies against bromelain‐treated mouse red blood cells (BrMRBC) and DNA) from different mouse strains did not contain any significant antibody activity against DNA and cytoskeleton proteins, although the presence of anti‐BrMRBC antibodies was clearly evident. Therefore, we investigated comparative natural antibody (NAb) specificities against an antigen panel (DNA, cytoskeleton proteins, IgG. bovine serum albumin (BSA), BrMRBC, trinitrophenyl (TNP), and trimethylammonium (TMA) haptens) among Ig‐secreting hybridoma collections from the splenic (158) and peritoneal (230) immune compartments of autoimmune New Zealand black (NZB) and lipopolysaccharide (LPS)‐stimulated BALB/c mice. The data showed: (i) isotypic restriction (μ and γ₃ only), predominance of TMA ion‐reactive (including BrMRBC) but negligible anti‐DNA‐reactive antibody specificities, and lack of simultaneous polyspecific widespread reactivity (i.e. at least four or more antigens) against DNA and cytoskeleton proteins in the peritoneal cavity; (ii) predominance of simultaneous widespread polyspecific reactivity against DNA and cytoskeleton proteins but negligible or no TMA hapten‐reactive antibody specificities in the spleen. These observations reflect certain differences in the B cell repertoire of peritoneal cavity (rich in Ly‐1 B cells) compared with spleen. The NAb against BrMRBC and those reactive with DNA and cytoskeleton proteins, which have been suggested to be secreted by the Ly‐1 B cell subset, are distinguishable on the basis of the presence of separate recurrent idiotypes and preferential localization of B lymphocytes directed against these autoantigens.