Identification of mouse CD4+ T cell epitopes in SARS-CoV-2 BA.1 spike and nucleocapsid for use in peptide:MHCII tetramers

Front Immunol. 2024 Mar 11:15:1329846. doi: 10.3389/fimmu.2024.1329846. eCollection 2024.

Abstract

Understanding adaptive immunity against SARS-CoV-2 is a major requisite for the development of effective vaccines and treatments for COVID-19. CD4+ T cells play an integral role in this process primarily by generating antiviral cytokines and providing help to antibody-producing B cells. To empower detailed studies of SARS-CoV-2-specific CD4+ T cell responses in mouse models, we comprehensively mapped I-Ab-restricted epitopes for the spike and nucleocapsid proteins of the BA.1 variant of concern via IFNγ ELISpot assay. This was followed by the generation of corresponding peptide:MHCII tetramer reagents to directly stain epitope-specific T cells. Using this rigorous validation strategy, we identified 6 immunogenic epitopes in spike and 3 in nucleocapsid, all of which are conserved in the ancestral Wuhan strain. We also validated a previously identified epitope from Wuhan that is absent in BA.1. These epitopes and tetramers will be invaluable tools for SARS-CoV-2 antigen-specific CD4+ T cell studies in mice.

Keywords: C57BL/6; HexaPro; epitope mapping; immunodominance; vFLIP.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD4-Positive T-Lymphocytes
  • COVID-19*
  • Epitopes, T-Lymphocyte
  • Histocompatibility Antigens Class II / chemistry
  • Mice
  • Nucleocapsid / chemistry
  • Peptides / chemistry
  • SARS-CoV-2* / chemistry
  • Spike Glycoprotein, Coronavirus / chemistry

Substances

  • Epitopes, T-Lymphocyte
  • Peptides
  • Histocompatibility Antigens Class II
  • spike protein, SARS-CoV-2
  • Spike Glycoprotein, Coronavirus