Comprehensive assessment of T-cell receptor β-chain diversity in αβ T cells
Comprehensive assessment of T-cell receptor β-chain diversity in αβ T cells
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DOI：
10.1182/blood-2009-04-217604
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发表时间：
2009-11-05
期刊：
影响因子：
20.3
通讯作者：
Carlson, Christopher S.
中科院分区：
文献类型：
--
作者：
Robins, Harlan S.;Campregher, Paulo V.;Carlson, Christopher S.
适应性免疫系统使用几种策略来产生具有足够多样性的T细胞和b细胞抗原受体库，以识别潜在病原体。在主要识别主要组织相容性复合体分子呈递的肽抗原的α - β T细胞中，这种受体多样性大部分包含在T细胞受体(TCR) α和β链的第三互补决定区（CDR3）中。尽管据估计适应性免疫系统可以产生多达10（16）对不同的α - β对，但TCR CDR3多样性的直接评估尚未证明适用于基于标准毛细管电泳的DNA测序。我们开发了一种新的实验和计算方法来测量基于单分子DNA测序的TCR CDR3多样性，并使用这种方法确定了来自2名成人T细胞的数百万重排TCR β基因的CDR3序列。我们发现总的TCR β受体多样性至少比以前的估计高4倍，CD45RO（+）抗原经历的α β T细胞亚群的多样性至少比以前的估计高10倍。这些方法对于在造血细胞移植后、先天性或获得性免疫缺陷状态以及正常衰老过程中评估α - β t细胞库多样性是有价值的。[血液。2009;114:4099-4107]
The adaptive immune system uses several strategies to generate a repertoire of T- and B-cell antigen receptors with sufficient diversity to recognize the universe of potential pathogens. In alpha beta T cells, which primarily recognize peptide antigens presented by major histocompatibility complex molecules, most of this receptor diversity is contained within the third complementarity-determining region (CDR3) of the T-cell receptor (TCR) alpha and beta chains. Although it has been estimated that the adaptive immune system can generate up to 10(16) distinct alpha beta pairs, direct assessment of TCR CDR3 diversity has not proved amenable to standard capillary electrophoresis-based DNA sequencing. We developed a novel experimental and computational approach to measure TCR CDR3 diversity based on single-molecule DNA sequencing, and used this approach to determine the CDR3 sequence in millions of rearranged TCR beta genes from T cells of 2 adults. We find that total TCR beta receptor diversity is at least 4-fold higher than previous estimates, and the diversity in the subset of CD45RO(+) antigen-experienced alpha beta T cells is at least 10-fold higher than previous estimates. These methods should prove valuable for assessment of alpha beta T-cell repertoire diversity after hematopoietic cell transplantation, in states of congenital or acquired immunodeficiency, and during normal aging. (Blood. 2009; 114: 4099-4107)