Luimstra Jolien J, Franken Kees L M C, Garstka Malgorzata A, Drijfhout Jan W, Neefjes Jacques, Ovaa Huib
Oncode Institute and Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands.
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
Curr Protoc Immunol. 2019 Sep;126(1):e85. doi: 10.1002/cpim.85.
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.
细胞毒性CD8 T细胞通过识别所有有核细胞上由MHC I类分子呈递的特定抗原介导细胞免疫。研究T细胞的相互作用和反应可为感染、自身免疫和癌症提供宝贵信息。荧光标记的MHC I多聚体能用于通过流式细胞术定量、表征和分离特定的CD8 T细胞。在此，我们描述了条件性MHC I多聚体的制备和应用，通过在特定温度下孵育，它们能够装载所选肽段。多聚体与模板肽折叠在一起，该模板肽在低温下形成稳定复合物，但在特定的升高温度下会解离。利用该技术可并行生成多个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使用时。© 2019作者。
