Abstract
The ongoing COVID-19 pandemic caused by SARS-CoV-2 has posed a devastating threat worldwide. The receptor-binding domain (RBD) of the spike protein is one of the most important antigens for SARS-CoV-2 vaccines, while the analysis of CD8 cytotoxic T lymphocyte activity in preclinical studies using mouse models is critical for evaluating vaccine efficacy. Here, we immunized C57BL/6 wild-type mice and transgenic mice expressing human angiotensin-converting enzyme 2 (ACE2) with the SARS-CoV-2 RBD protein to evaluate the IFN-γ-producing T cells in the splenocytes of the immunized mice using an overlapping peptide pool by an enzyme-linked immunospot assay and flow cytometry. We identified SARS-CoV-2 S395–404 as a major histocompatibility complex (MHC) class I-restricted epitope for the RBD-specific CD8 T cell responses in C57BL/6 mice.
Highlights
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative pathogen of human coronavirus disease 2019 (COVID-19) [1,2]
One week after the last immunization, an IFN-γ Enzyme-Linked Immunospot (ELISPOT) assay was performed by stimulating the splenocytes obtained from immunized mice with an receptor-binding domain (RBD) peptide pool of 42 synthetic RBD peptides with 15-mers overlapping by 5 aa (Table 1)
The level of increase provoked by treatment with the S391–405 peptide was similar to the spot forming units (SFUs) values achieved via stimulation with the peptide pool (214 ± 95 SFUs) (p < 0.0001)
Summary
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative pathogen of human coronavirus disease 2019 (COVID-19) [1,2]. Most COVID-19 vaccine candidates have focused on the SARS-CoV-2 spike protein or the receptor-binding domain (RBD) of the spike protein as the vaccine antigen [4]. The SARS-CoV-2 RBD primarily binds to angiotensin-converting enzyme 2 (ACE2). Facilitates the entry of the virus into host cells [5]. CD8 cytotoxic T lymphocytes (CTLs) play a critical role in eliminating virus-infected cells, thereby leading to the effective clearance of viral infection. CTL activity is important to provide long-term protection from reinfection of the virus [6]
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