SARS-CoV-2 PepTivator Peptide Pools can be used to investigate T cell immunity in COVID-19 after natural infection or vaccination. Load antigen-presenting cells with the peptides and stimulate SARS-CoV-2–specific T cells. The T cells can then be detected and/or isolated for further research, including analysis of T cell immune response after vaccination or monitoring the immune status after infection. We provide different peptide pools covering specified proteins parts of wild-type and mutated SARS-CoV-2 variants.
Application of SARS-CoV-2 PepTivator Peptide Pools
- Individuals recovered from COVID-19 show elevated levels of IFN-γ–producing CD3+ T cells upon stimulation with SARS-CoV-2 PepTivator Peptide Pools
Stimulation with SARS-CoV-2 PepTivator Peptide Pools shows the presence of virus-specific T cells in individuals recovered from COVID-19. After stimulation with the peptide pools, intracellular IFN-γ was stained and analyzed in CD3+ T cells using flow cytometry.
Samples of a healthy donor and a recovered COVID-19 patient were stimulated for 4 h with the indicated SARS-CoV-2 PepTivator Peptide Pools (Prot_N, Prot_M, Prot_S, or a mix of all three) in the presence of BFA. As negative control, samples were left untreated without (w/o) antigen. Subsequently, T cell lineage surface markers and intracellular cytokines were stained. The presented plots are exemplary data showing IFN-γ+CD3+ T cells. The data demonstrate an increased number of activated T cells in recovered COVID-19 patients upon stimulation with the SARS-CoV-2 peptide pools. Quantitative analysis is shown below.
Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use. - Quantitative analysis of spike protein–specific CD4+ T cells in convalescent COVID-19 donors
Both options stimulate SARS-CoV-2–reactive T cells to a similar extend as shown by the exemplary analysis of SARS-CoV-2–specific CD4+ T cells in recovered individuals and are suitable for analyzing the immune response upon vaccination with a vaccine based on the spike protein.
Samples of a healthy donor and a recovered COVID-19 patient were stimulated with SARS-CoV-2 PepTivator Peptide Pools (Prot_N, Prot_M, Prot_S, or a mix of all three) and examined for IFN-γ production. Stained cells were analyzed using flow cytometry and the frequencies of IFN-γ+CD3+ T cells are shown in the bar chart. Flow data are shown above.
Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use. - Detecting virus-specific T cells after COVID-19 vaccination
Spike protein–reactive T cells were stimulated with PepTivator SARS-CoV-2 Prot_S Complete, which covers the complete sequence of the mature spike protein. Analysis of T cell activation markers illustrates that this PepTivator Peptide Pool is suitable for stimulation of virus-specific CD4+ and CD8+ T cells after COVID-19 vaccination.
Samples of a healthy donor and a recovered COVID-19 patient were stimulated with SARS-CoV-2 PepTivator Peptide Pools (mix of Prot_N, Prot_M and Prot_S), left untreated without (w/o) antigens as negative control, or incubated with influenza antigens as positive control. Based on the Rapid ARTE protocol, the cells were subsequently stained (CD3, CD4, CD8, CD154, and TNF-α) and enriched. Cells were gated on CD4+ cells for the presented exemplary flow data. Analysis of CD154 and TNF-α shows successful enrichment of rare SARS-CoV-2–specific CD4+ T cells.
Data are kindly provided by Alexander Scheffold and Petra Bacher, UKSH, Kiel, Germany.
Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use.
SARS‑CoV‑2 PepTivator® Peptide Pools Product Information:
| PepTivator SARS-CoV-2 Prot_S: | |||
| 130-126-700 | PepTivator SARS-CoV-2 Prot_S, 6 nmol per peptide | for stimulation of 1×10^8 cells | The PepTivator SARS-CoV-2 Prot_S covers selected immunodominant sequence domains of the spike protein (aa 304–338, 421–475, 492–519, 683–707, 741–770, 785–802, and 885–1273). |
| 130-126-701 | PepTivator SARS-CoV-2 Prot_S, 60 nmol per peptide | for stimulation of 1×10^9 cells | |
| PepTivator SARS-CoV-2 Prot_S1: | |||
| 130-127-041 | PepTivator SARS-CoV-2 Prot_S1, 6 nmol per peptide | for stimulation of 1×10^8 cells | SARS-CoV-2 Prot_S1 covers the complete N-terminal S1 domain of the spike protein (aa 1–692) |
| 130-127-048 | PepTivator SARS-CoV-2 Prot_S1, 60 nmol per peptide | for stimulation of 1×10^9 cells | |
| PepTivator SARS-CoV-2 Prot_S+: | |||
| 130-127-311 | PepTivator SARS-CoV-2 Prot_S+, 6 nmol per peptide | for stimulation of 1×10^8 cells | SARS-CoV-2 Prot_S+ parts of the C-terminal S2 domain (aa 689–895) |
| 130-127-312 | PepTivator SARS-CoV-2 Prot_S+, 60 nmol per peptide | for stimulation of 1×10^9 cells | |
| PepTivator SARS-CoV-2 Prot_N: | |||
| 130-126-698 | PepTivator SARS-CoV-2 Prot_N, 6 nmol per peptide | for stimulation of 1×10^8 cells | covering the complete sequence of the nucleocapsid phosphoprotein (“N”) of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43423.2) |
| 130-126-699 | PepTivator SARS-CoV-2 Prot_N, 60 nmol per peptide | for stimulation of 1×10^9 cells | |
| PepTivator SARS-CoV-2 Prot_M: | |||
| 130-126-702 | PepTivator SARS-CoV-2 Prot_M, 6 nmol per peptide | for stimulation of 1×10^8 cells | covering the complete sequence of the nucleocapsid phosphoprotein (“N”) of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43423.2). |
| 130-126-703 | PepTivator SARS-CoV-2 Prot_M, 60 nmol per peptide | for stimulation of 1×10^9 cells | |
Schematic alignment of the spike glycoprotein and the SARS‑CoV‑2 PepTivators Prot_S, Prot_S1, and Prot_S+:
PepTivator SARS‑CoV‑2 Prot_S covers the predicted immunodominant domains of the SARS‑CoV‑2 spike glycoprotein (protein S), PepTivator SARS‑CoV‑2 Prot_S1 covers the N-terminal S1 domain and PepTivator SARS‑CoV‑2 Prot_S+ covers a part of the C-terminal S2 domain.