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.
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.
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发表时间：
1998-09
影响因子：
4.4
通讯作者：
W. Ikematsu;J. Kobarg;H. Ikematsu;Y. Ichiyoshi;P. Casali
中科院分区：
文献类型：
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作者：
W. Ikematsu;J. Kobarg;H. Ikematsu;Y. Ichiyoshi;P. Casali
在先前的工作中，我们使用来自接种灭活狂犬病病毒（胸腺依赖性（TD）嵌合抗原）的4名受试者的B细胞，制备了4种IgM、5种IgG 1和1种IgA 1抗狂犬病病毒的mAb，并对mAb V（H）DJ（H）基因进行了测序。在这里，我们克隆了V κ J κ和V λ J λ基因，以完成这些单克隆抗体的Ag结合位点的一级结构。虽然抗狂犬病病毒单克隆抗体选择VA基因（2e.2.2两次，DPL 11和DPL 23）反映了人类单倍体基因组中V λ基因的代表性（随机利用），但V κ基因（O2/O 12两次，O 8/O 18，A3/A19，A27和L2）的代表性（p = 0.0018）（非随机利用）。此外，抗狂犬病病毒单克隆抗体对V κ和V λ基因的选择与557种不同的V κ J κ重排、253种γ-球蛋白型丙种球蛋白病中的V λ J λ重排以及其他抗胸腺依赖性Ag的单克隆抗体，包括23种抗HIV单克隆抗体和51种类风湿因子，但与43株抗流感嗜血杆菌B型多糖（一种原型胸腺非依赖性（TI）Ag）的Ab不同。抗狂犬病病毒mAb V κ J κ和V λ J λ片段显示可变数目的体细胞突变，在mAb 58和病毒中和mAb 57中，这在互补决定区中引起显著浓度的氨基酸置换（分别为p = 0.0028和p = 0.0023），表明Ag的选择。这种Ag依赖的体细胞选择过程叠加在发生在用于Ag重排的B细胞受体阶段的体细胞多样化过程上，并且需要V基因3'截短和N核苷酸添加以产生异质性CDR 3。
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.