Open AccessProduction and Thermal Exchange of Conditional Peptide‐MHC I Multimers
Author(s) -
Luimstra Jolien J.,
Franken Kees L. M. C.,
Garstka Malgorzata A.,
Drijfhout Jan W.,
Neefjes Jacques,
Ovaa Huib
Publication year - 2019
Publication title -
current protocols in immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.618
H-Index - 43
eISSN - 1934-368X
pISSN - 1934-3671
DOI - 10.1002/cpim.85
Subject(s) - cytotoxic t cell , mhc class i , cd8 , major histocompatibility complex , flow cytometry , t cell , microbiology and biotechnology , biology , antigen , mhc restriction , chemistry , immunology , immune system , biochemistry , in vitro
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.
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