CLONAL ANALYSIS OF A HUMAN-ANTIBODY RESPONSE - III - NUCLEOTIDE-SEQUENCES OF MONOCLONAL IGM, IGG, AND IGA TO RABIES VIRUS REVEAL RESTRICTEDVK GENE UTILIZATION, JUNCTIONAL V-KAPPA-J-KAPPA AND V-LAMBDA-J-LAMBDADIVERSITY, AND SOMATIC HYPERMUTATION

In previous work, me generated four IgM, five IgG1, and one IgA1 mAbs to rabies virus using B cells from four subjects vaccinated with inact ivated 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-bi nding site of these mAbs, While the anti-rabies virus mAb selection of V lambda genes (2e.2.2 twice, DPL11, and DPL23) reflected the represe ntation 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,vith that of 557 assorted V kappa J kappa rearrangements, that of 253 V lambda J lambda rearrangements in lambd a-type gammopathies, and that of other Abs to thymus-dependent Ags, in cluding 23 anti-HIV mAbs and 51 rheumatoid factors, but differed from that of 43 Abs to Haemophilus influenzae type b polysaccharide, a prot otypic thymus-independent (TI) Ag, The anti-rabies virus mAb V kappa J kappa and V lambda J lambda segments displayed variable numbers of so matic mutations, which, in mAb58 and the virus-neutralizing mAb57, ent ailed a significant concentration of amino acid replacements in the co mplementarity-determining regions (p = 0.0028 and p = 0.0023, respecti vely), suggesting a selection by Ag. This Ag-dependent somatic selecti on process was superimposed on a somatic diversification process that occurred at the stage of B cell receptor for Ag rearrangement, and tha t entailed V gene 3' truncation and N nucleotide additions to yield he terogeneous CDR3s.