Cukalac Tania, Kan Wan-Ting, Dash Pradyot, Guan Jing, Quinn Kylie M, Gras Stephanie, Thomas Paul G, La Gruta Nicole L
Department of Microbiology and Immunology, University of Melbourne, Peter Doherty Institute of Infection and Immunity, Melbourne, VIC, Australia.
Department of Immunology, St Jude Children's Research Hospital, Memphis, TN, USA.
Immunol Cell Biol. 2015 Oct;93(9):804-14. doi: 10.1038/icb.2015.44. Epub 2015 Mar 25.
T-cell receptor (TCR) usage has an important role in determining the outcome of CD8(+) cytotoxic T-lymphocyte responses to viruses and other pathogens. However, the characterization of TCR usage from which such conclusions are drawn is based on exclusive analysis of either the TCRα chain or, more commonly, the TCRβ chain. Here, we have used a multiplexed reverse transcription-PCR protocol to analyse the CDR3 regions of both TCRα and β chains from single naive or immune epitope-specific cells to provide a comprehensive picture of epitope-specific TCR usage and selection into the immune response. Analysis of TCR repertoires specific for three influenza-derived epitopes (D(b)NP(366), D(b)PA(224) and D(b)PB1-F2(62)) showed preferential usage of particular TCRαβ proteins in the immune repertoire relative to the naive repertoire, in some cases, resulting in a complete shift in TRBV preference or CDR3 length, and restricted repertoire diversity. The NP(366)-specific TCRαβ repertoire, previously defined as clonally restricted based on TCRβ analysis, was similarly diverse as the PA(224)- and PB1-F2(62)-specific repertoires. Intriguingly, preferred TCR characteristics (variable gene usage, CDR3 length and junctional gene usage) appeared to be able to confer specificity either independently or in concert with one another, depending on the epitope specificity. These data have implications for established correlations between the nature of the TCR repertoire and response outcomes after infection, and suggest that analysis of a subset of cells or a single TCR chain does not accurately depict the nature of the antigen-specific TCRαβ repertoire.
T细胞受体（TCR）的使用在决定CD8⁺细胞毒性T淋巴细胞对病毒和其他病原体的反应结果中起着重要作用。然而，得出此类结论所依据的TCR使用特征是基于对TCRα链或更常见的TCRβ链的单独分析。在此，我们使用了一种多重逆转录PCR方案来分析单个初始或免疫表位特异性细胞的TCRα和β链的互补决定区3（CDR3）区域，以全面了解表位特异性TCR的使用情况以及其在免疫反应中的选择。对三种流感衍生表位（D(b)NP(366)、D(b)PA(224)和D(b)PB1-F2(62)）特异性的TCR库分析表明，相对于初始库，免疫库中特定的TCRαβ蛋白有优先使用情况，在某些情况下，导致TRBV偏好或CDR3长度发生完全转变，并且库的多样性受限。先前基于TCRβ分析被定义为克隆受限的NP(366)特异性TCRαβ库，与PA(224)和PB1-F2(62)特异性库同样具有多样性。有趣的是，根据表位特异性，偏好的TCR特征（可变基因使用、CDR3长度和连接基因使用）似乎能够独立或相互协同赋予特异性。这些数据对已确立的TCR库性质与感染后反应结果之间的相关性具有启示意义，并表明对细胞亚群或单个TCR链的分析不能准确描绘抗原特异性TCRαβ库的性质。
