Clonal analysis of a human antibody response. III. Nucleotide sequences of monoclonal IgM, IgG, and IgA to rabies virus reveal restricted V kappa gene utilization, junctional V kappa J kappa and V lambda J lambda diversity, and somatic hypermutation.
In previous work, we generated four IgM, five IgG1, and one IgA1 mAbs to rabies virus using B cells from four subjects vaccinated with inactivated rabies virus, a thymus-dependent (TD) mosaic Ag, and sequenced the mAb V(H)DJ(H) genes. Here, we have cloned the V kappa J kappa and V lambda J lambda genes to complete the primary structure of the Ag-binding site of these mAbs. While the anti-rabies virus mAb selection of VA genes (2e.2.2 twice, DPL11, and DPL23) reflected the representation of the V lambda genes in the human haploid genome (stochastic utilization), that of V kappa genes (O2/O12 twice, O8/O18, A3/A19, A27, and L2) did not (p = 0.0018) (nonstochastic utilization). Furthermore, the selection of both V kappa and V lambda genes by the anti-rabies virus mAbs vastly overlapped with that of 557 assorted V kappa J kappa rearrangements, that of 253 V lambda J lambda rearrangements in lambda-type gammopathies, and that of other Abs to thymus-dependent Ags, including 23 anti-HIV mAbs and 51 rheumatoid factors, but differed from that of 43 Abs to Haemophilus influenzae type b polysaccharide, a prototypic thymus-independent (TI) Ag. The anti-rabies virus mAb V kappa J kappa and V lambda J lambda segments displayed variable numbers of somatic mutations, which, in mAb58 and the virus-neutralizing mAb57, entailed a significant concentration of amino acid replacements in the complementarity-determining regions (p = 0.0028 and p = 0.0023, respectively), suggesting a selection by Ag. This Ag-dependent somatic selection process was superimposed on a somatic diversification process that occurred at the stage of B cell receptor for Ag rearrangement, and that entailed V gene 3' truncation and N nucleotide additions to yield heterogeneous CDR3s.
Ikematsu
W
W
Department of Pathology, Cornell University Medical College, New York 10021, USA.
Kobarg
J
J
Ikematsu
H
H
Ichiyoshi
Y
Y
Casali
P
P
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D000916:Antibody Diversity / Q000235:genetics; D001483:Base Sequence; D002999:Clone Cells; D003001:Cloning, Molecular; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D005803:Genes, Immunoglobulin; D015483:HIV Antibodies / Q000737:chemistry / Q000235:genetics; D019741:Haemophilus influenzae type b / Q000276:immunology; D006801:Humans; D007070:Immunoglobulin A / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007133:Immunoglobulin Joining Region / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007075:Immunoglobulin M / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010265:Paraproteinemias / Q000235:genetics / Q000276:immunology; D010446:Peptide Fragments / Q000235:genetics; D011135:Polysaccharides, Bacterial / Q000235:genetics / Q000276:immunology; D011820:Rabies virus / Q000276:immunology*; D012217:Rheumatoid Factor / Q000737:chemistry / Q000235:genetics
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015483:HIV Antibodies; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007133:Immunoglobulin Joining Region; D007075:Immunoglobulin M; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D010446:Peptide Fragments; D011135:Polysaccharides, Bacterial; D012217:Rheumatoid Factor