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

tetano

Editor, Senior Moderator
Front Immunol


. 2024 Mar 11:15:1329846.
doi: 10.3389/fimmu.2024.1329846. eCollection 2024. Identification of mouse CD4[SUP]+[/SUP] T cell epitopes in SARS-CoV-2 BA.1 spike and nucleocapsid for use in peptide:MHCII tetramers

Laura Bricio-Moreno[SUP] 1 2 3 [/SUP], Juliana Barreto de Albuquerque[SUP] 1 2 3 [/SUP], Jake M Neary[SUP] 1 2 [/SUP], Thao Nguyen[SUP] 1 2 [/SUP], Lucy F Kuhn[SUP] 1 2 [/SUP], YeePui Yeung[SUP] 1 2 [/SUP], Kathryn M Hastie[SUP] 4 [/SUP], Sara Landeras-Bueno[SUP] 4 [/SUP], Eduardo Olmedillas[SUP] 4 [/SUP], Chitra Hariharan[SUP] 4 [/SUP], Anusha Nathan[SUP] 5 6 [/SUP], Matthew A Getz[SUP] 5 [/SUP], Alton C Gayton[SUP] 5 [/SUP], Ashok Khatri[SUP] 3 7 [/SUP], Gaurav D Gaiha[SUP] 3 5 8 [/SUP], Erica Ollmann Saphire[SUP] 4 9 [/SUP], Andrew D Luster[SUP] 1 2 3 [/SUP], James J Moon[SUP] 1 2 3 10 [/SUP]



Affiliations
Abstract

Understanding adaptive immunity against SARS-CoV-2 is a major requisite for the development of effective vaccines and treatments for COVID-19. CD4[SUP]+[/SUP] 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[SUP]+[/SUP] T cell responses in mouse models, we comprehensively mapped I-A[SUP]b[/SUP]-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[SUP]+[/SUP] T cell studies in mice.

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

 
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