Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a γ2a variable region
Copyright policy )Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a γ2a variable region
To examine germ line genes of the heavy chain variable region (VH) that might contribute to formation of antibodies of the NPb family, we have derived cDNA clones from two hybridomas making NPb antibodies. One, B1-8, made an IgM protein and was derived during a primary response; the other, S43, made an IgG2a protein and was derived during a hyperimmune response. Sequence comparison of the two clones showed that they differed by only 10 bp in the VH region, had very different D segments and had identical J segments (J2). A set of closely related germ line VH genes was then cloned from a partial Eco RI library of C57Bl/6 DNA. By comparing the germ line VH regions to the cDNA VH regions, we identified seven potential candidates for encoding the VH regions of NPb antibodies. The seven VH regions were sequenced, and one V(186-2) contained exactly the DNA sequence found in the clone derived from B1-8. None of the DNA sequence differences that distinguished the S43-derived clone from the B1-8 clone was found in any of the other six germ line genes. Because the S43 sequence was more closely related to the V(186-2) germ line sequence than to any of the other VH genes, we conclude that the differences between the genes resulted from somatic mutation and that the two hybridomas derived their VH regions from the same germ line gene. Certain of the sequenced VH genes contain crippling mutations; the repertoire of germ line VH genes that can contribute to the diversity of antibodies may therefore be less than the total number of genes detectable by hydridization.
- Massachusetts Institute of Technology United States
- University of Cologne Germany
Base Sequence, Immunoglobulin gamma-Chains, Genes, MHC Class II, Immunoglobulin Variable Region, Antibodies, Monoclonal, DNA, Antibodies, Nitrophenols, Mice, Mutation, Animals, Binding Sites, Antibody, Cloning, Molecular, Immunoglobulin Heavy Chains, Haptens, Antibody Diversity, Phenylacetates
Base Sequence, Immunoglobulin gamma-Chains, Genes, MHC Class II, Immunoglobulin Variable Region, Antibodies, Monoclonal, DNA, Antibodies, Nitrophenols, Mice, Mutation, Animals, Binding Sites, Antibody, Cloning, Molecular, Immunoglobulin Heavy Chains, Haptens, Antibody Diversity, Phenylacetates
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