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

STAR Protoc. 2021-08; 
Chanchan Xiao, Congling Qiu, Jieping Deng, Jiezhou Ye, Lijuan Gao, Jun Su, Oscar Junhong Luo, Pengcheng Wang, Guobing Chen
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Peptide Synthesis … The candidate peptides were synthesized in GenScript Biotechnology Co., Ltd (Nanjing, China) and dissolved in DMSO or in water with the … The candidate peptides (Table 1) are synthesized in GenScript Biotechnology and dissolved in DMSO at the concentration of 10 mM. … Get A Quote

摘要

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).

关键词

Cell Biology, Cell isolation, Immunology, Microbiology