CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
英文原题:Assessment of human and mouse tumor-antigen specific CD8 + T cells by multimer staining in multiple compartments.
Assessment of human and mouse tumor-antigen specific CD8 + T cells by multimer staining in multiple compartments.
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在肿瘤部位及外周检测肿瘤抗原特异性T细胞,对于研究和监测直接参与抗肿瘤免疫应答的T细胞至关重要。T细胞通过其T细胞受体(TCR)识别由主要组织相容性复合体(MHC)分子呈递肿瘤抗原肽的肿瘤细胞。可利用与荧光染料偶联的多重肽-MHC分子,通过流式细胞术分析抗原特异性CD8+和CD4+ T细胞。这些肽-MHC多聚体结合流式细胞术的持续进展,改变了抗原特异性T细胞的表征方式,可用于感染、自身免疫和癌症研究中T细胞的可视化、计数、追踪、表型鉴定和分离,也可用于监测多种疗法过程中的特异性适应性免疫应答。本文详细介绍一种简便且准确的多聚体染色方案,用于检测从人和小鼠肿瘤及外周血单个核细胞(PBMC)中分离的肿瘤抗原特异性CD8+ T细胞。
Detection of tumor antigen-specific T cells at the tumor site and in the periphery is pivotal to study and monitor T cells directly involved in the antitumor immune response. Through their T-cell receptor (TCR), T cells recognize tumor cells presenting their antigenic peptides by major histocompatibility complex (MHC) molecules. Analyses of antigen-specific CD8 + and CD4 + T cells can be realized by flow cytometry, using fluorochrome-conjugated multiplexed peptide-MHC molecules.
These peptide-MHC multimers, in conjunction with continuing advances in flow cytometry, have transformed the characterization of antigen-specific T cells. They enable the visualization, enumeration, tracking, phenotype characterization and isolation of T cells during infection, autoimmunity and cancer. They can be useful to monitor specific adaptive responses during various type of therapies.
Here we detail a simple and accurate protocol of multimer staining of tumor antigen-specific CD8 + T cells isolated from human and mouse tumors and Peripheral Blood Mononuclear Cells (PBMCs).
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