Department of Microbiology and Immunology, The University of Melbourne, Parkville, Victoria 3052, Australia.
Nat Commun. 2013;4:2333. doi: 10.1038/ncomms3333.
T cell repertoire diversity and clonotype follow-up in vaccination, cancer, infectious and immune diseases represent a major challenge owing to the enormous complexity of the data generated. Here we describe a next generation methodology, which combines 5'RACE PCR, 454 sequencing and, for analysis, IMGT, the international ImMunoGeneTics information system (IMGT), IMGT/HighV-QUEST web portal and IMGT-ONTOLOGY concepts. The approach is validated in a human case study of T cell receptor beta (TRB) repertoire, by chronologically tracking the effects of influenza vaccination on conventional and regulatory T cell subpopulations. The IMGT/HighV-QUEST paradigm defines standards for genotype/haplotype analysis and characterization of IMGT clonotypes for clonal diversity and expression and achieves a degree of resolution for next generation sequencing verifiable by the user at the sequence level, while providing a normalized reference immunoprofile for human TRB.
T 细胞受体库多样性和克隆型分析在疫苗接种、癌症、感染和免疫性疾病方面极具挑战性，这主要是由于产生的数据极其复杂。在这里，我们描述了一种下一代方法，该方法结合了 5'RACE PCR、454 测序，以及用于分析的 IMGT，即国际免疫遗传学信息系统 (IMGT)、IMGT/HighV-QUEST 网页门户和 IMGT-ONTOLOGY 概念。该方法在人类 T 细胞受体 β (TRB) 受体库的案例研究中得到了验证，通过按时间顺序跟踪流感疫苗对常规和调节性 T 细胞亚群的影响。IMGT/HighV-QUEST 范例为基因型/单倍型分析以及 IMGT 克隆型的特征定义了标准，用于克隆多样性和表达的分析，并实现了用户在序列水平上可验证的下一代测序的分辨率，同时为人类 TRB 提供了标准化的参考免疫分析。
