高通量测序分析人类TCRβ链CDR3的 repertoire 特征

Analysis of the Repertoire Features of TCR Beta Chain CDR3 in Human by High-Throughput Sequencing.

作者信息

Hou Xianliang, Wang Mingbang, Lu Chong, Xie Qian, Cui Guangying, Chen Jianing, Du Yu, Dai Yong, Diao Hongyan

机构信息

State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.

出版信息

Cell Physiol Biochem. 2016;39(2):651-67. doi: 10.1159/000445656. Epub 2016 Jul 21.

Abstract

BACKGROUND/AIMS: To ward off a wide variety of pathogens, the human adaptive immune system harbors a vast array of T-cell receptors, collectively referred to as the TCR repertoire. Assessment of the repertoire features of TCR is vital for us to deeper understand of immune behaviour and immune response.

METHODS

In this study, we used a combination of multiplex-PCR, Illumina sequencing and IMGT (ImMunoGeneTics)/HighV-QUEST for a standardized analysis of the repertoire features of TCR beta chain in the blood of healthy individuals, including the repertoire features of public TCR complementarity-determining regions (CDR3) sequences, highly expanded clones, long TCR CDR3 sequences.

RESULTS

We found that public CDR3 sequences and high-frequency sequences had the same characteristics, both of them had fewer nucleotide additions and shorter CDR3 length, which were closer to the germline sequence. Moreover, our studies provided evidence that public amino acid sequences are produced by multiple nucleotide sequences. Notably, there was skewed VDJ segment usage in long CDR3 sequences, the expression levels of 10 TRβV segments, 7 TRβJ segments and 2 TRβD segments were significantly different in the long CDR3 sequences compared to the short CDR3 sequences. Moreover, we identified that extensive N additions and increase of D gene usage contributing to TCR CDR3 length, and observed there was distinct usage frequency of amino acids in long CDR3 sequences compared to the short CDR3 sequences.

CONCLUSIONS

Some repertoire features could be observed in the public sequences, highly abundance clones, and long TCR CDR3 sequences, which might be helpful for further study of immune behavior and immune response.

摘要

背景/目的:为抵御多种病原体,人类适应性免疫系统拥有大量的T细胞受体,统称为TCR库。评估TCR库的特征对于我们深入了解免疫行为和免疫反应至关重要。

方法

在本研究中,我们采用多重PCR、Illumina测序和IMGT(免疫遗传学)/HighV-QUEST相结合的方法,对健康个体血液中TCRβ链的库特征进行标准化分析,包括公共TCR互补决定区(CDR3)序列的库特征、高度扩增的克隆、长TCR CDR3序列。

结果

我们发现公共CDR3序列和高频序列具有相同的特征,它们的核苷酸添加较少,CDR3长度较短,更接近胚系序列。此外,我们的研究提供了证据表明公共氨基酸序列由多个核苷酸序列产生。值得注意的是,长CDR3序列中VDJ片段的使用存在偏倚,与短CDR3序列相比,长CDR3序列中10个TRβV片段、7个TRβJ片段和2个TRβD片段的表达水平存在显著差异。此外,我们确定广泛的N添加和D基因使用的增加导致TCR CDR3长度增加,并观察到与短CDR3序列相比,长CDR3序列中氨基酸的使用频率明显不同。

结论

在公共序列、高丰度克隆和长TCR CDR3序列中可以观察到一些库特征,这可能有助于进一步研究免疫行为和免疫反应。

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