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STAR Protocols

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Optimization of antigen-specific CD8+ T?cell activation conditions for infectious diseases including COVID-19

Published:17 September 2021 DOI: 10.1016/j.xpro.2021.100789 PMID: 34414379
Chanchan Xiao, Congling Qiu, Jieping Deng, Jiezhou Ye, Lijuan Gao, Jun Su, Oscar Junhong Luo, Pengcheng Wang, Guobing Chen

Abstract

Here, we describe the use of the artificial antigen-presenting cell (aAPC) system for the verification of T-cell epitopes. We purify and activate CD8+ T?cells from blood samples from HLA-A2 that are negative for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). CD8+ T?cells are combined with peptide-loaded T2-A2 cells, which are then stained with a SARS-CoV-2-specific MHC-1 tetramer to identify specific HLA-A2-restricted T-cell epitopes. The use of aAPC and healthy donors means that only BSL2 lab conditions are needed. For details of the use and implementation of this protocol, please refer to Deng et al. (2021).

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