Production and purification of infectious diseases-associated tetramer for staining of CD8+ T cells from PBMCs

Jiezhou Ye; Wen Lei; ChanChan Xiao; Xiangmeng Su; Jie Gao; Xinran Li; Wenxin Li; Pengcheng Wang; Zhenyou Jiang; Guobing Chen

STAR Protocols (Mar 2022)

Production and purification of infectious diseases-associated tetramer for staining of CD8+ T cells from PBMCs

Jiezhou Ye,
Wen Lei,
ChanChan Xiao,
Xiangmeng Su,
Jie Gao,
Xinran Li,
Wenxin Li,
Pengcheng Wang,
Zhenyou Jiang,
Guobing Chen

Affiliations

Jiezhou Ye: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China; Guangdong-Hong Kong-Macau Great Bay Area Geroscience Joint Laboratory, Guangzhou, China
Wen Lei: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China; Guangdong-Hong Kong-Macau Great Bay Area Geroscience Joint Laboratory, Guangzhou, China
ChanChan Xiao: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China; Guangdong-Hong Kong-Macau Great Bay Area Geroscience Joint Laboratory, Guangzhou, China
Xiangmeng Su: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China
Jie Gao: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China
Xinran Li: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China
Wenxin Li: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China
Pengcheng Wang: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China
Zhenyou Jiang: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China; Corresponding author
Guobing Chen: Department of Microbiology and Immunology, Institute of Geriatric Immunology, School of Medicine, Jinan University, Guangzhou, China; Guangdong-Hong Kong-Macau Great Bay Area Geroscience Joint Laboratory, Guangzhou, China; Affiliated Huaqiao Hospital, Jinan University, Guangzhou, China; Corresponding author

Journal volume & issue: Vol. 3, no. 1
p. 101206

Abstract

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Summary: Major histocompatibility complex (MHC) tetramers can work as diagnostic tools to identify antigen-specific T cells in immunological research and monitoring. Here, we provide a general protocol for the production of MHC tetramer. We obtain highly pure N-terminal His-tagged HLA-A2 α chain and β2-microglobulin (β2m) to fold a monomer with a photocleavable peptide, which can exchange with an HLA-A2 presented peptide derived from influenza A virus. Further those monomers compose tetramer to stain antigen-specific CD8+ T cells.For complete details on the use and execution of this protocol, please refer to Xiao C.C. et al. (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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