TCR Repertoire Analysis by Next Generation Sequencing Allows Complex Differential Diagnosis of T Cell-Related Pathology
TCR Repertoire Analysis by Next Generation Sequencing Allows Complex Differential Diagnosis of T Cell-Related Pathology
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DOI：
10.1111/ajt.12431
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发表时间：
2013-11-01
影响因子：
8.8
通讯作者：
Babel, N.
中科院分区：
文献类型：
--
作者：
Dziubianau, M.;Hecht, J.;Babel, N.
克隆型分析对于抗原特异性T细胞的完整表征是必不可少的。此外，关于克隆身份的知识允许追踪全血和组织浸润中的抗原特异性T细胞，并且可以提供关于抗原特异性的信息。在这里，我们开发了基于下一代测序（NGS）的平台，用于T细胞的高度定量克隆型表征，并确定了输入材料（DNA、RNA、离体衍生或细胞培养扩增的T细胞）的无偏表征的要求。此后，我们进行了T细胞受体（TCR）的各种标本在临床设置，包括巨细胞病毒（CMV），多瘤病毒BK（BKV）的再活化和急性细胞移植排斥反应的库分析。我们的研究结果揭示了病毒特异性T细胞克隆型的动态性质; CMV再激活与新的高度丰富的抗原特异性克隆的出现有关。此外，BKV，同种异体抗原特异性T细胞，肾移植和尿来源的淋巴细胞之间的克隆型重叠的分析提供了提示同种异体移植功能障碍的鉴别诊断，并使适当的治疗调整。我们相信，所建立的方法将提供深入了解病毒特异性/抗肿瘤免疫的调节，并具有较高的诊断潜力，在临床routine.This研究描述了开发和表征的T细胞受体库分析的基础上下一代测序及其应用复杂的鉴别诊断移植后肾功能不全。
Clonotype analysis is essential for complete characterization of antigen-specific T cells. Moreover, knowledge on clonal identity allows tracking of antigen-specific T cells in whole blood and tissue infiltrates and can provide information on antigenic specificity. Here, we developed a next generation sequencing (NGS)-based platform for the highly quantitative clonotype characterization of T cells and determined requirements for the unbiased characterization of the input material (DNA, RNA, ex vivo derived or cell culture expanded T cells). Thereafter we performed T cell receptor (TCR) repertoire analysis of various specimens in clinical settings including cytomegalovirus (CMV), polyomavirus BK (BKV) reactivation and acute cellular allograft rejection. Our results revealed dynamic nature of virus-specific T cell clonotypes; CMV reactivation was linked to appearance of new highly abundant antigen-specific clonalities. Moreover, analysis of clonotype overlap between BKV-, alloantigen-specific T cell-, kidney allograft- and urine-derived lymphocytes provided hints for the differential diagnosis of allograft dysfunction and enabled appropriate therapy adjustment. We believe that the established approach will provide insights into the regulation of virus-specific/anti-tumor immunity and has high diagnostic potential in the clinical routine.This study describes the development and characterization of T cell receptor repertoire analysis based on Next Generation Sequencing and its application for complex differential diagnosis of posttransplant kidney dysfunction.