Next-generation sequencing reveals new insights about gene usage and CDR-H3 composition in the horse antibody repertoire
Next-generation sequencing reveals new insights about gene usage and CDR-H3 composition in the horse antibody repertoire
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DOI：
10.1016/j.molimm.2018.11.017
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发表时间：
2019-01-01
影响因子：
3.6
通讯作者：
Felicori, Liza F.
中科院分区：
文献类型：
--
作者：
Manso, Taciana Conceicao;Groenner-Penna, Michele;Felicori, Liza F.
一个多世纪以来，马血清抗体一直被用于治疗和预防传染病和中毒。然而，对产生马血清抗体的免疫遗传多样性知之甚少。在这里，我们采用下一代测序对马b细胞库进行了首次全面分析。重链（IGH）和轻链（IGL）基因座分别获得了近45,000和30,000个克隆型。我们观察到常见亚群IGHV2（92.49%）和IGLV8（82.50%）的倾斜使用，与先前的报道一致，但也有罕见基因IGHV6S1和IGLV4S2的新使用。与人、兔和小鼠相比，CDR-H3环的氨基酸组成在106和116位显示出不同的氨基酸模式，这表明马的CDR-H3环的构象主要是延长的。我们的分析为马体内产生抗体多样性的机制提供了新的见解，并可适用于未来治疗用途的合成抗体的优化设计。
Horse serum antibodies have been used for greater than a century for the treatment and prophylaxis of infectious diseases and envenomations. Little is known, however, about the immunogenetic diversity that produces horse serum antibodies. Here, we employed next-generation sequencing for a first-in-kind comprehensive analysis of the equine B-cell repertoire. Nearly 45,000 and 30,000 clonotypes were obtained for the heavy-chain (IGH) and lambda light-chain (IGL) loci, respectively. We observed skewed use of the common subgroups IGHV2 (92.49%) and IGLV8 (82.50%), consistent with previous reports, but also novel use of the rare genes IGHV6S1 and IGLV4S2. CDR-H3 amino acid composition revealed different amino acid patterns at positions 106 and 116 compared to human, rabbit, and mouse, suggesting that an extended conformation predominates among horse CDR-H3 loops. Our analysis provides new insights regarding the mechanisms employed to generate antibody diversity in the horse, and could be applicable to the optimized design of synthetic antibodies intended for future therapeutic use.