Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.
Department of Medicine Huddinge, Center for Infectious Medicine (CIM), Karolinska Institutet, Stockholm, Sweden.
Methods Mol Biol. 2022;2574:209-219. doi: 10.1007/978-1-0716-2712-9_9.
T-cell repertoire characterization is a methodology that enables the identification of T-cell receptor (TCR) gene sequences in a T-cell population. TCR genes are composed of modular gene segments V (D) J that undergo somatic recombination, resulting in unique and unpredictable sequences that can be utilized to identify each T-cell clone. The analysis of the TCR composition in a T-cell population can give information on the biological phenomenon such as antigen-driven expansion and heterogeneity of T-cell responses. Bulk TCR analysis can give useful information on the clonality and can help track a specific clonotype over time or in different compartments, although the information about pairing cannot be provided. Single-cell TCR sequencing, on the other hand, can provide pairing information that are necessary to reconstruct the TCR and confirm antigen specificity.Here we describe common methods to characterize T-cell repertoires based on both bulk and single-cell next-generation sequencing.
T 细胞受体(TCR)基因序列的分析方法可用于鉴定 T 细胞群体中的 TCR 基因序列。TCR 基因由模块化的基因片段 V(D)J 组成,这些片段经历体细胞重组,产生独特且不可预测的序列,可用于鉴定每个 T 细胞克隆。分析 T 细胞群体中的 TCR 组成可以提供关于生物学现象的信息,例如抗原驱动的扩增和 T 细胞反应的异质性。批量 TCR 分析可以提供关于克隆性的有用信息,并有助于跟踪特定克隆型随时间或在不同隔室中的变化,尽管无法提供配对信息。相比之下,单细胞 TCR 测序可以提供配对信息,这些信息对于重建 TCR 和确认抗原特异性是必要的。在这里,我们描述了基于批量和单细胞下一代测序来描述 T 细胞受体库的常见方法。