Background information
SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) was first detected in December 2019 in Wuhan, China. Investigations of the cellular immune response revealed the crucial role of virus-reactive T cells after natural infection and vaccination. Analysis of antigen-specific CD8+ T cells has, however, proven challenging for certain individuals due to low T cell frequencies. To efficiently stimulate SARS-CoV-2–reactive CD8+ T cells, PepTivator SARS-CoV-2 MHC-I Select and SARS-CoV-2 MHC-I Select Prot_S were designed.
PepTivator SARS-CoV-2 MHC-I Select consists of 173 MHC class I–restricted peptides of 8–11 aa length. Sequences are derived from the following structural and non-structural SARS-CoV-2 proteins: envelope protein, membrane protein, nucleoprotein, ORF1ab, ORF3a, ORF7a, ORF8, and ORF10 (GenBank MN908947.3). The peptides are restricted to the following HLA-molecules: HLA‑A*01, A*02, A*03, A*11, A*24, A*26, A*29, A*30, A*31, A*33, A*68,B*07, B*08, B*13, B*15, B*27, B*35, B*39, B*40, B*44, B*51, B*54, B*57, B*58, C*07.
PepTivator SARS-CoV-2 MHC-I Select Prot_S consists of 146 MHC class I–restricted peptides of 8–12 aa length derived exclusively from the SARS-CoV-2 surface or spike glycoprotein (“S”) (GenBank MN908947.3, Protein QHD43416.1).The peptides are restricted to the following HLA-molecules: HLA‑A*01, A*02, A*03, A*11, A*24, A*26, A*29, A*30, A*31, A*32, A*68, B*07, B*08, B*14, B*15, B*27, B*35, B*39, B*40, B*44, B*51, B*53, B*54, B*57, B*58, C*07.
The MHC-I–specific SARS-CoV-2 PepTivators can be
- • applied individually to analyze a) CD8+ T cells reactive to the SARS-CoV-2 spike protein (PepTivator SARS-CoV-2 MHC-I Select Prot_S) or b) CD8+ T cells reactive to the whole SARS-CoV-2 proteome without the spike protein (PepTivator SARS-CoV-2 MHC-I Select)
- • used together (PepTivator SARS-CoV-2 MHC-I Select plus PepTivator SARS-CoV-2 MHC-I Select Prot_S) to analyze CD8+ T cells reactive to the whole SARS-CoV-2 proteome, or
- • used to supplement other PepTivators to boost the CD8+ T cell response and analyze CD4+ as well as CD8+ T cells.
Comparison of T cell reactivity towards the spike protein and other SARS-CoV-2 proteins allows to distinguish vaccine-induced from natural T cell immunity.
Applications
The in vitro stimulation of SARS-CoV-2–specific CD8+ T cells with PepTivators causes secretion of effector cytokines and upregulation of activation markers, which then allow the detection and isolation of SARS-CoV-2–specific T cells:
- Detection and analysis of antigen-specific CD8+ effector/memory T cells in PBMCs by MACS Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
- Isolation of viable antigen-specific CD8+ T cells using MACS Cytokine Secretion Assay – Cell Enrichment and Detection Kits. Subsequently, cells can be expanded for generation of T cell lines.
- Generation antigen-specific CD8+ effector/memory T cells from naive T cell populations.
- Pulsing of antigen-presenting cells, e.g. for research on dendritic cell vaccination.
- Detection and analysis of antigen-specific CD8+ effector/memory T cells in PBMCs by MACS Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
- Isolation of viable antigen-specific CD8+ T cells using MACS Cytokine Secretion Assay – Cell Enrichment and Detection Kits. Subsequently, cells can be expanded for generation of T cell lines.
- Generation antigen-specific CD8+ effector/memory T cells from naive T cell populations.
- Pulsing of antigen-presenting cells, e.g. for research on dendritic cell vaccination.
Overview
Stimulate CD8+ T cells reactive to SARS-CoV-2 using PepTivator Peptide Pools. MHC-I specific PepTivators are pools of lyophilized peptides, consisting of MHC class I-restricted peptides of 8–12 aa length. PepTivator SARS-CoV-2 MHC-I Select covers selected epitopes of the whole proteome of SARS-CoV-2 (GenBank MN908947.3) without the surface or spike glycoprotein (“S”). In contrast, PepTivator SARS-CoV-2 MHC-I Select Prot_S covers selected epitopes of the SARS-CoV-2 surface or spike glycoprotein (“S”) (GenBank MN908947.3, Protein QHD43416.1).
In vitro stimulation of antigen-specific T cells with PepTivator Peptide Pools causes the secretion of effector cytokines and the up-regulation of activation markers, which then allow the detection and isolation of antigen-specific T cells.
