Generation of high-affinity antibody against T cell-dependent antigen in the Ganp gene-transgenic mouse
Generation of high-affinity antibody against T cell-dependent antigen in the Ganp gene-transgenic mouse
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DOI：
10.4049/jimmunol.174.8.4485
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发表时间：
2005-04-15
影响因子：
4.4
通讯作者：
Kuwahara, K
中科院分区：
文献类型：
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作者：
Sakaguchi, N;Kimura, T;Kuwahara, K
在B细胞中缺乏生发中心相关DNA引发酶（GANP）的小鼠中，高亲和力Ab的产生受损。在这项研究中，我们研究了其过表达在ganp转基因C57 BL/6小鼠（Ganp（Tg））中的作用。Ganp（Tg）在B细胞发育、血清IG水平和对T细胞非依赖性Ag的应答中显示正常表型;然而，与C57 BL/6相比，其产生的Ab对硝基苯鸡丙种球蛋白具有更高的亲和力。为了进一步检查亲和力增加，我们建立了产生高亲和力mAb的杂交瘤，并使用BIAcore比较它们的亲和力。C57 BL/6产生了高亲和力的抗硝基苯基单克隆抗体（K-D类似于2.50 × 10（-7）M）的IgG 1/λ 1，并含有具有W33 L突变的V（H）186.2区。GanpTg通过使用V（H）186.2以及非典型V（H）7183区域产生高得多的亲和力（K-D > 1.57 × 10（-9）M）。Ganp（Tg）还产生了具有中和活性的异常高亲和力的抗HIV-1（V3肽）mAb（K，> 9.90 x 10-11 m）。这些结果表明，GANP参与V区改变，产生高亲和力Ab。
Generation of high-affinity Ab is impaired in mice lacking germinal center-associated DNA primase (GANP) in B cells. In this study, we examined the effect of its overexpression in ganp transgenic C57BL/6 mice (Ganp(Tg)). Ganp(Tg) displayed normal phenotype in B cell development, serum Ig levels, and responses against T cell-independent Ag; however, it generated the Ab with much higher affinity against nitrophenyl-chicken gammaglobulin in comparison with C57BL/6. To further examine the affinity increase, we established hybridomas producing high-affinity mAbs and compared their affinities using BIAcore. C57BL/6 generated high-affinity anti-nitrophenyl mAbs (K-D similar to 2.50 X 10(-7) M) of IgG1/lambda 1 and contained the V(H)186.2 region with W33L mutation. GanpTg generated much higher affinity (K-D > 1.57 X 10(-9) M) by usage of V(H)186.2 as well as noncanonical V(H)7183 regions. Ganp(Tg) also generated exceptionally high-affinity anti-HIV-1 (V3 peptide) mAbs (K, > 9.90 x 10-11 m) with neutralizing activity. These results demonstrated that GANP is involved in V region alteration generating high-affinity Ab.