Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization.

Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization.
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DOI:
10.1038/nbt.2593
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发表时间:
2013-07
影响因子:
46.9
通讯作者:
--
中科院分区:
工程技术1区
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--
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目前还不可能预测不同个体的T细胞会识别哪些表位。这是彻底分析和理解接种疫苗或感染后t细胞反应的障碍。在这里,通过结合大量细胞术和组合肽- mhc四聚体染色,我们开发了一种方法,可以快速和同时识别和表征许多表位特异性的T细胞。我们使用它在单个人类血液样本中筛选多达109种不同的肽- mhc四聚体,同时仍然保留至少23个标记来分析t细胞表型和功能的其他标记。在77个轮状病毒候选表位中,我们在健康人血液中鉴定出6个限制于人白细胞抗原(HLA)-A*0201的t细胞表位。轮状病毒VP3蛋白表位特异性T细胞表现出独特的表型,并在肠上皮中高频存在。这种方法对于t细胞对传染病或疫苗的反应的综合分析应该是有用的。
It is currently not possible to predict which epitopes will be recognized by T cells in different individuals. This is a barrier to the thorough analysis and understanding of T-cell responses after vaccination or infection. Here, by combining mass cytometry with combinatorial peptide–MHC tetramer staining, we have developed a method allowing the rapid and simultaneous identification and characterization of T cells specific for many epitopes. We use this to screen up to 109 different peptide–MHC tetramers in a single human blood sample, while still retaining at least 23 labels to analyze other markers of T-cell phenotype and function. Among 77 candidate rotavirus epitopes, we identified six T-cell epitopes restricted to human leukocyte antigen (HLA)-A*0201 in the blood of healthy individuals. T cells specific for epitopes in the rotavirus VP3 protein displayed a distinct phenotype and were present at high frequencies in intestinal epithelium. This approach should be useful for the comprehensive analysis of T-cell responses to infectious diseases or vaccines.
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