Alanio Cécile, Bouvier Isabelle, Jusforgues-Saklani Hélène, Albert Matthew L
Immunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d'Immunologie Humaine, Institut Pasteur, Paris, France.
Methods Mol Biol. 2013;960:309-326. doi: 10.1007/978-1-62703-218-6_23.
The tracking of epitope-specific T cells is a useful approach for the study of adaptive immune responses. This protocol describes how Major Histocompatibility Complex Class I (MHC-I) multimers can be used to stain, enrich, and enumerate (rare) populations of CD8(+) T cells specific for a given antigen. It provides the detailed steps for multimer labeling, magnetic enrichment, and cytometric analysis. Additionally, it provides informations for multiplexing experiments in order to achieve simultaneous detection of multiple antigenic specificities, and strategies for coupling the protocol with functional assays (e.g., intracellular cytokine staining). Future developments in cytometric systems (e.g., mass spectroscopy-based cytometry) and gene expression studies (e.g., single cell PCR) will extend these approaches and provide an unprecedented assessment of the immune repertoire.
对表位特异性T细胞进行追踪是研究适应性免疫反应的一种有用方法。本方案描述了如何使用主要组织相容性复合体I类（MHC-I）多聚体来染色、富集和计数针对给定抗原的CD8(+) T细胞（稀有）群体。它提供了多聚体标记、磁性富集和细胞计数分析的详细步骤。此外，它还提供了用于多重实验以实现同时检测多种抗原特异性的信息，以及将该方案与功能测定（如细胞内细胞因子染色）相结合的策略。细胞计数系统（如基于质谱的细胞计数）和基因表达研究（如单细胞PCR）的未来发展将扩展这些方法，并对免疫库进行前所未有的评估。
