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

STAR Protoc. 2021 Sep 17;2(3):100789. doi: 10.1016/j.xpro.2021.100789. Epub 2021 Aug 14.

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).

Keywords: Cell Biology; Cell isolation; Immunology; Microbiology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD8-Positive T-Lymphocytes / immunology*
  • COVID-19 / immunology*
  • Epitopes, T-Lymphocyte / immunology*
  • HLA-A2 Antigen / immunology*
  • Humans
  • Lymphocyte Activation
  • SARS-CoV-2 / immunology*
  • Spike Glycoprotein, Coronavirus / immunology*
  • Tumor Necrosis Factor Receptor Superfamily, Member 7 / immunology*

Substances

  • Epitopes, T-Lymphocyte
  • HLA-A2 Antigen
  • Spike Glycoprotein, Coronavirus
  • Tumor Necrosis Factor Receptor Superfamily, Member 7