Development and characterization of an anti-idiotype antibody to the capsular polysaccharide of Neisseria meningitidis serogroup C

Abstract

A monoclonal anti-idiotypic antibody (Ab2) whose antibody combining site contained a surrogate image of the meningococcal group C capsular polysaccharide was developed. To accomplish this, a monoclonal antibody against the group C capsular polysaccharide was developed by the fusion of splenocytes from mice immunized with Neisseria meningitidis group C strain MP13 with Sp2/0-Ag14 plasmacytoma cells. Monoclonal antibody 1E4, an immunoglobulin M isotype, demonstrated binding to the serogroup C polysaccharide in enzyme-linked immunosorbent assay (ELISA). Monoclonal antibody 1E4 reacted with 30 of 30 group C strains and 1 of 36 group B strains in immunodot assay, slide agglutination, inhibition ELISA, and bactericidal assay. This monoclonal antibody was selected as idiotype (Ab1) for the development of hybridomas producing an anti-idiotype antibody. One of the hybridomas developed, designated 6F9, was capable of over 70% inhibition of 1E4 in binding in the meningococcal C polysaccharide-specific ELISA. Studies with convalescent human serum demonstrated 100% inhibition of a serogroup C-specific ELISA with 200 micrograms of 6F9 per ml and 50% inhibition of this ELISA was achieved with 50 micrograms of 6F9 per ml. Monoclonal anti-idiotype antibodies (Ab3) with specificities similar to Ab1, 1E4 were generated from BALB/c mice immunized with the Ab2 (6F9). Immunization of rabbits with 6F9 resulted in an immunoglobulin G response which was significantly greater than that of control to a titer of 1:160. These studies indicate that monoclonal 6F9 contained a surrogate image on the combining antibody site which mimicked meningococcal C polysaccharide. This surrogate image is capable of evoking antibodies to the meningococcal C polysaccharide in syngenic and xenogenic species.