Production and Thermal Exchange of Conditional Peptide-MHC I Multimers.
Production and Thermal Exchange of Conditional Peptide-MHC I Multimers.
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DOI：
10.1002/cpim.85
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发表时间：
2019-09-01
影响因子：
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通讯作者：
Ovaa, Huib
中科院分区：
文献类型：
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作者：
Luimstra, Jolien J;Franken, Kees L M C;Ovaa, Huib
细胞毒性 CD8+ T 细胞通过识别所有有核细胞上 MHC I 类呈递的特定抗原来介导细胞免疫。研究 T 细胞相互作用和反应可以提供有关感染、自身免疫和癌症的宝贵信息。荧光标记的 MHC I 多聚体可用于通过流式细胞术定量、表征和分离特定的 CD8+ T 细胞。在这里，我们描述了条件 MHC I 多聚体的生产和使用，这些多聚体可以通过在规定的温度下孵育来装载所选的肽。多聚体与模板肽折叠，在低温下形成稳定的复合物，但在指定的高温下解离。使用该技术可以并行生成多个 MHC I 多聚体，以便对大量抗原特异性 CD8+ T 细胞进行染色和分离，特别是与条形码技术结合时。 © 2019 作者。
Cytotoxic CD8+ T cells mediate cellular immunity through recognition of specific antigens presented by MHC class I on all nucleated cells. Studying T cell interactions and responses provides invaluable information on infection, autoimmunity and cancer. Fluorescently labeled multimers of MHC I can be used to quantify, characterize, and isolate specific CD8+ T cells by flow cytometry. Here we describe the production and use of conditional MHC I multimers that can be loaded with peptides of choice by incubating them at a defined temperature. Multimers are folded with a template peptide that forms a stable complex at low temperature, but dissociates at a defined elevated temperature. Using this technology multiple MHC I multimers can be generated in parallel, to allow staining and isolation of large sets of antigen-specific CD8+ T cells, especially when combined with barcoding technologies. © 2019 The Authors.