VH and VL gene usage by anti-beta-amyloid autoantibodies in Alzheimer's disease: detection of highly mutated V regions in both heavy and light chains.
In a previous study, four human IgM kappa monoclonal antibodies (mAbs) secreted by Epstein-Barr-virus-transformed B cell lines independently derived from the peripheral blood of a patient with Alzheimer's disease (AD) were found to react with a conformational epitope of beta-amyloid protein and to stain amyloid plaques in AD brain. Three of these mAbs were studied further. They did not react with an additional panel of antigens and autoantigens. By a competitive inhibition ELISA, the Kd was determined to be 5.7 x 10(-8) M. The VH and V kappa of these mAbs were sequenced at the cDNA level and found to be identical. The corresponding VH and V kappa germline counterpart genes hsigghvm148 and hsiggkvm148 were identified to be analogous to the previous reported VH3 germline gene humighvf1/dp53 and the V kappa IV germline gene hsigk18. DA1/4 and JH4b were used by the heavy chain and J kappa 4 was used by the light chain. Multiple nucleotide substitutions were seen in both VH and V kappa when their sequences were compared to the germline sequences. High replacement/silent ratios in the CDR regions of both VH and V kappa indicate positive-selective pressure. Of a total of 22 amino acid replacements in VH and V kappa, 12 were nonconservative replacements. Furthermore, 7 of these 12 resulted in charge changes. The monoreactivity, the moderately high affinity constant, the clonal expansion evident by identical VH and VL used by these B cells secreting antibodies of interest, and the excessive replacement mutations in both VH and VL segments lead to the conclusion that these antibodies have been generated due to an antigen-driven process.
Fang
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Department of Medicine, University of Virginia School of Medicine, Charlottesville 22908.
Kannapell
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Fu
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Xu
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Gaskin
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D000544:Alzheimer Disease / Q000235:genetics / Q000276:immunology*; D016229:Amyloid beta-Peptides / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D000940:Antigenic Variation; D001323:Autoantibodies / Q000235:genetics*; D001483:Base Sequence; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation
D016229:Amyloid beta-Peptides; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains