Abstract
IntroductionSimilar to antibody detection, severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2)-specific T-cell response evaluation is also pivotal among the coronavirus disease 2019 (COVID-19) convalescents and the vaccinated populations. Nucleocapsid (N) protein is one of the main structural proteins of SARS-CoV-2 and can trigger T-cell responses in humans.MethodsAn overlapping peptide pool covering the full length of the N protein was designed, peptides with positive T-cell activating potency in COVID-19 convalescents were screened, and CD8+ T cell epitopes were further identified. The epitope was used to detect the SARS-CoV-2-specific CD8+ T cell responses in COVID-19 convalescents based in intracellular cytokine staining and tetramer staining in flow cytometry. ResultsA human leukocyte antigen A (HLA-A)*1101-restricted CD8+ T cell epitope, which could stimulate the production of IFN-γ via peripheral blood mononuclear cells (PBMCs) of the convalescents was defined, and the tetramer generated with this epitope could detect SARS-CoV-2-specific T cells in the PBMCs of the convalescents. The structural investigation eliminated that the epitope was a typical HLA-A*1101-restricted T-cell epitope which was conserved among all the sarbecoviruses. DiscussionThe newly identified SARS-CoV-2-derived T-cell epitope was helpful to detect the cellular immunity against different sarbecoviruses including SARS-CoV and SARS-CoV-2. This study provided an evaluation method and also a peptide candidate for the research and development of T-cell based vaccine for the virus.
Highlights
Similar to antibody detection, severe acute respiratory syndrome coronavirus type 2 (SARSCoV-2)-specific T-cell response evaluation is pivotal among the coronavirus disease 2019 (COVID19) convalescents and the vaccinated populations
CD8+ T cells recognize antigenic peptides presented by major histocompatibility complex I (MHC-I) through T cell receptors (TCRs) and are activated to kill virusinfected target cells [4,5]
Through enzyme-linked immunospot assay (ELISpot), a human leukocyte antigen A (HLAA)*1101-restricted CD8+ T cell epitope, which could stimulate the production of IFN-γ by the peripheral blood mononuclear cells (PBMCs) of the convalescents, was defined, and the tetramer generated with this epitope could detect SARS-CoV-2-specific T cells in the PBMCs of the convalescents
Summary
Severe acute respiratory syndrome coronavirus type 2 (SARSCoV-2)-specific T-cell response evaluation is pivotal among the coronavirus disease 2019 (COVID19) convalescents and the vaccinated populations. Nucleocapsid (N) protein is one of the main structural proteins of SARS-CoV-2 and can trigger T-cell responses in humans. The in vitro refolding experiments and crystallographic structural analysis showed that the epitope was a typical HLAA*1101-restricted and conserved among all the sarbecoviruses. These findings provided the basis for cellular immunity evaluation in COVID-19 recovered patients and vaccinated donors, and the newly identified epitope was helpful for the development of a polypeptide vaccine for SARS-CoV-2
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