Specific H chain junctional diversity may be required for non-T15 antibodies to bind phosphorylcholine.
The secondary antibody response of mice to phosphorylcholine (PC) shows a markedly different clonal profile than the primary response. In particular, the T15 antibodies that dominate the primary response are a minor part of secondary IgG antibodies, whereas 511 and 603 antibodies become a more prominent part of the PC-specific secondary response. These three anti-PC families differ only in L chain usage. We partially sequenced the IgH chain mRNA of a series of secondary T15 and 511 hybridomas to determine the role of somatic mutation and affinity maturation in these changes in clonal profile. None of the sequenced T15 antibodies showed somatic mutations or affinity increases. In contrast, all of the 511 antibodies had extensive somatic mutation and most had significantly increased affinity for nitrophenyl-PC. The failure of T15-expressing B cells to contribute to the secondary IgG response thus is likely to be explained by their inability to undergo (or tolerate) substantial somatic mutation and affinity maturation. We also noted that all 511 antibodies sequenced by us or others had an extra amino acid encoded at the VH-D junction by either N region addition or diversity of VH-D joining. Published sequences also show a 603 family-specific change at the VH-D junction. The frequency with which these changes, which appear obligate for PC binding, occur may determine the under-representation of these clonotypes in the primary anti-PC response. The affinity maturation in 511 antibodies after somatic mutation appears to account for their expansion in the secondary response.
D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D000916:Antibody Diversity; D000918:Antibody Specificity; D001483:Base Sequence; D002794:Choline / Q000031:analogs & derivatives*; D005260:Female; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000302:isolation & purification; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D007133:Immunoglobulin Joining Region / Q000235:genetics* / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D008805:Mice, Inbred A; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D009154:Mutation; D009194:Myeloma Proteins / Q000276:immunology* / Q000378:metabolism; D010767:Phosphorylcholine / Q000276:immunology* / Q000378:metabolism
D007143:Immunoglobulin Heavy Chains; D007130:Immunoglobulin Idiotypes; D007133:Immunoglobulin Joining Region; D007135:Immunoglobulin Variable Region; D009194:Myeloma Proteins; C043925:myeloma protein TEPC15; D010767:Phosphorylcholine; D002794:Choline