Laboratory of Molecular and Cellular Biology, Graduate School of Science and Engineering for Research, University of Toyama, Toyama-shi, Toyama, Japan.
Medical & Biological Laboratories Co., Ltd., Ina-shi, Nagano, Japan.
PLoS One. 2018 Dec 13;13(12):e0208977. doi: 10.1371/journal.pone.0208977. eCollection 2018.
The guinea pig has been used as a model to study various human infectious diseases because of its similarity to humans regarding symptoms and immune response, but little is known about the humoral immune response. To better understand the mechanism underlying the generation of the antibody repertoire in guinea pigs, we performed deep sequencing of full-length immunoglobulin variable chains from naïve B and plasma cells. We gathered and analyzed nearly 16,000 full-length VH, Vκ and Vλ genes and analyzed V and J gene segment usage profiles and mutation statuses by annotating recently reported genome data of guinea pig immunoglobulin genes. We found that approximately 70% of heavy, 73% of kappa and 81% of lambda functional germline V gene segments are integrated into the actual V(D)J recombination events. We also found preferential use of a particular V gene segment and accumulated mutation in CDRs 1 and 2 in antigen-specific plasma cells. Our study represents the first attempt to characterize sequence diversity in the expressed guinea pig antibody repertoire and provides significant insight into antibody repertoire generation and Ig-based immunity of guinea pigs.
豚鼠因其在症状和免疫反应方面与人类相似,被用作研究各种人类传染病的模型,但人们对体液免疫反应知之甚少。为了更好地了解豚鼠抗体库产生的机制,我们对幼稚 B 细胞和浆细胞中的全长免疫球蛋白可变链进行了深度测序。我们收集并分析了近 16000 个全长 VH、Vκ 和 Vλ 基因,并通过注释最近报道的豚鼠免疫球蛋白基因的基因组数据,分析了 V 和 J 基因片段的使用情况和突变状态。我们发现,大约 70%的重链、73%的κ链和 81%的λ链功能性胚系 V 基因片段整合到实际的 V(D)J 重组事件中。我们还发现,在抗原特异性浆细胞中,特定 V 基因片段的使用和 CDR1 和 CDR2 中的突变积累存在偏好性。我们的研究代表了首次尝试描述表达的豚鼠抗体库中的序列多样性,为豚鼠抗体库产生和基于 Ig 的免疫提供了重要的见解。