ANTIGEN-BINDING AND IDIOTYPE ANALYSIS OF ANTIBODIES OBTAINED AFTER ELECTROPORATION OF HEAVY AND LIGHT CHAIN GENES ENCODING PHOSPHOCHOLINE-SPECIFIC ANTIBODIES - A MODEL FOR T15-IDIOTYPE DOMINANCE

Antibodies bearing the T15 idiotype dominate the murine primary immune response to phosphocholine (PC). Analysis of antigen binding of antibodies derived from V1:DFL16.1:J(H)1 (V(H)1) germline and N region-derived variant heavy (H) chains and kappa22, kappa24, and kappa8 light (L) chains demonstrates that the T15H:kappa22L (T15) antibody binds PC at least 20-40 times better than other antibodies derived from alternate germline forms of the V(H)1 H chain and kappa22, kappa24, or kappa8 L chains. To achieve affinities in the same range as the T15 antibody, kappa24 and kappa8 L chain-containing antibodies must have H chains derived from variant N region or somatically mutated V(H)1 genes. Single amino acid differences at the VD junction of the various germline and N region variant V(H)1 H chains dictate the L chain that can associate with the H chain to produce a PC-specific antibody. Several H:L combinations give rise to T15 or M167 idiotype-positive antibodies that lack specificity for PC, and single amino acid substitutions or insertions at the V(H)1:D junction result in the loss of T15 or M167 idiotopes. Based on these observations, our data support a molecular model involving both preferential gene rearrangement and antigen-driven B cell selection to explain T15 idiotype dominance in the immune response to PC. In the absence of N region diversification, large numbers of neonatal B cells bearing the T15H:kappa22L surface immunoglobulin M (sIgM) receptors would be selected and expanded by autologous or environmental PC antigen into the long-lived peripheral B cell pool.