PepTivator® SARS-CoV-2 Prot_N

PepTivator
®
SARS-CoV-2 Prot_N is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the complete sequence of the nucleocapsid phosphoprotein (“N”) of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43423.2).
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.

Data and images for
PepTivator
®
SARS-CoV-2 Prot_N

Figures

Figure 1

View details
Exemplary analysis of SARS-CoV-2–specific CD4+ and CD8+ T cells.
A) PBMCs were stimulated with a mix of PepTivator SARS-CoV-2 Prot_N, Prot_M, and Prot_S or unstimulated as negative control. The data shows CD154 and TNF-α for CD4
+
T cells and TNF-α and IFN-γ for CD8
+
T cells. B/C). Either PepTivators covering the complete sequence of the nucleoprotein (N; PepTivator SARS-CoV-2 Prot_N), the membrane protein
(M; PepTivator SARS-CoV-2 Prot_M), the spike protein (S; PepTivator SARS-CoV-2 Prot_S, Prot_S1, and Prot_S+), or the mix of all were used to stimulate SARS-CoV-2–reactive T cells. Frequencies of CD154+ within CD4
+
T cells and TNF-α
+
within CD8
+
T cells are shown. In the quantitative analysis (B) each dot corresponds to one donor and in the heat maps (C) SARS-CoV-2–reactive T cell frequencies measured for each donor are color coded.

Figure 1

Exemplary analysis of SARS-CoV-2–specific CD4+ and CD8+ T cells.
A) PBMCs were stimulated with a mix of PepTivator SARS-CoV-2 Prot_N, Prot_M, and Prot_S or unstimulated as negative control. The data shows CD154 and TNF-α for CD4
+
T cells and TNF-α and IFN-γ for CD8
+
T cells. B/C). Either PepTivators covering the complete sequence of the nucleoprotein (N; PepTivator SARS-CoV-2 Prot_N), the membrane protein
(M; PepTivator SARS-CoV-2 Prot_M), the spike protein (S; PepTivator SARS-CoV-2 Prot_S, Prot_S1, and Prot_S+), or the mix of all were used to stimulate SARS-CoV-2–reactive T cells. Frequencies of CD154+ within CD4
+
T cells and TNF-α
+
within CD8
+
T cells are shown. In the quantitative analysis (B) each dot corresponds to one donor and in the heat maps (C) SARS-CoV-2–reactive T cell frequencies measured for each donor are color coded.

Figure 2

View details
Recovered COVID-19 patients show elevated levels of IFN-γ–producing CD3+ T cells upon stimulation with SARS‑CoV‑2 PepTivator Peptide Pools.
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.

Figure 2

Recovered COVID-19 patients show elevated levels of IFN-γ–producing CD3+ T cells upon stimulation with SARS‑CoV‑2 PepTivator Peptide Pools.
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.

Specifications for
PepTivator
®
SARS-CoV-2 Prot_N

Overview

PepTivator
®
SARS-CoV-2 Prot_N is a pool of lyophilized peptides, consisting mainly of 15-mer sequences with 11 amino acids overlap, covering the complete sequence of the nucleocapsid phosphoprotein (“N”) of SARS-Coronavirus 2 (GenBank MN908947.3, Protein QHD43423.2).
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.

Detailed product information

Background information

SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), also known as novel coronavirus 2019-nCoV, causes fever, severe respiratory illness and can lead to life threatening pneumonia. The first cases of this disease, termed COVID-19 for coronavirus disease 2019, have been detected in December 2019 Wuhan, China.
SARS-CoV-2 Prot_N stands for the nucleocapsid phosphoprotein of SARS-CoV-2. This protein is responsible for the packaging of the genome of coronaviruses and, thus plays a role during replication, transcription as well as virion assembly. Therefore, the nucleocapsid phosphoprotein (“N”) of SARS-CoV-2 has been suggested as possible target for vaccine development.

Applications

PepTivator
®
SARS-CoV-2 Prot_N – research grade has been specifically developed for
in vitro
stimulation of SARS-CoV-2–specific T cells. Peptides of 15 amino acids in length and 11 amino acids overlap represent an optimized solution for stimulating both CD4
+
and CD8
+
T cells in various applications, including:
  • Detection and analysis of SARS-CoV-2–specific CD4+ and CD8+ effector/memory T cells in PBMCs by MACS® Cytokine Secretion Assays, intracellular cytokine staining, or other technologies.
  • Isolation of viable SARS-CoV-2–specific CD4+ T cells with the CD154 MicroBead Kit, or of viable CD4+ and 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 of SARS-CoV-2–specific CD4+ and CD8+ effector/ memory T cells from naive T cell populations.
  • Pulsing of antigen-presenting cells e.g. for research on dendritic cell vaccination.
More information regarding the work with antigen-specific T cells can be found in the T cell application section.

References for
PepTivator
®
SARS-CoV-2 Prot_N

Publications

  1. Sattler, A. et al. (2020) SARS-CoV-2-specific T cell responses and correlations with COVID-19 patient predisposition J. Clin. Invest. 130(12): 6477-6489
  2. Bacher, P. et al. (2020)
    Low avidity CD4
    +
    T cell responses to SARS-CoV-2 in unexposed individuals and humans with severe COVID-19
    Immunity 53: 1258-1271
  3. Bonifacius, A. et al. (2021) COVID-19 immune signatures reveal stable antiviral T cell function despite declining humoral responses Immunity 54(2): 340-354
  4. Shomuradova, A. S. et al. (2020) SARS-CoV-2 epitopes are recognized by a public and diverse repertoire of human T cell receptors. Immunity 53(6): 1245-1257
  5. Habel, J. R. et al. (2020)
    Suboptimal SARS-CoV-2-specific CD8
    +
    T cell response associated with the prominent HLA-A*02:01 phenotype
    Proc. Natl. Acad. Sci. U.S.A. 117(3): 24384-24391
  6. Albecka, A. et al. A functional assay for serum detection of antibodies against SARS-CoV-2 nucleoprotein EMBO J. (DOI: 10.15252/embj.2021108588)
  7. Sekine, T. et al. (2020) Robust T Cell Immunity in Convalescent Individuals with Asymptomatic or Mild COVID-19 Cell 183(1): 158-168
  8. Au, L. et al. (2021) Cytokine release syndrome in a patient with colorectal cancer after vaccination with BNT162b2 Nat Med 27(8): 1362-1366
  9. Ishigaki, H. et al. (2021) Neutralizing antibody-dependent and -independent immune responses against SARS-CoV-2 in cynomolgus macaques Virology 554: 97-105
  10. Sir Karakus, G. et al. (2021) Preclinical efficacy and safety analysis of gamma-irradiated inactivated SARS-CoV-2 vaccine candidates Sci Rep 11(1): 5804
  11. Cooper, R. S. et al. (2021) Rapid GMP-Compliant Expansion of SARS-CoV-2-Specific T Cells From Convalescent Donors for Use as an Allogeneic Cell Therapy for COVID-19 Front Immunol 11: 598402
  12. Strafella, C. et al. (2021) Case Report: Sars-CoV-2 Infection in a Vaccinated Individual: Evaluation of the Immunological Profile and Virus Transmission Risk Front Immunol (doi: 10.3389/fimmu.2021.708820)
  13. Chan, Y-H. et al. Asymptomatic COVID-19: disease tolerance with efficient anti-viral immunity against SARS-CoV-2 EMBO Mol. Med. (doi: 10.15252/emmm.202114045)
  14. Anft, M. et al. (2020) COVID-19 progression is potentially driven by T cell immunopathogenesis medRxiv - the preprint server for health sciences (doi.org/10.1101/2020.04.28.20083089 )
  15. Fendler, A. et al. (2020) Adaptive immunity to SARS-CoV-2 in cancer patients: The CAPTURE study medRxiv - the preprint server for health sciences : doi.org/10.1101/2020.12.21.20248608
  16. Grau-Expósito, J. et al. Peripheral and lung resident T cell responses against SARS-CoV-2 medRxiv - the preprint server for health sciences : doi.org/10.1101/2020.12.02.20238907
  17. Jung, J. H. et al. (2021) SARS-CoV-2-specific T Cell Memory is Sustained in COVID-19 Convalescents for 8 Months with Successful Development of Stem Cell-like Memory T Cells medRxiv - the preprint server for health sciences (doi.org/10.1101/2021.03.04.21252658)
  18. Leung, W. et al. Successful manufacturing of clinical-grade SARS-CoV-2–specific T cells for adoptive cell therapy. medRxiv - the preprint server for health sciences : doi.org/10.1101/2020.04.24.20077487
  19. Riou, C. et al. (2020) Rapid, simplified whole blood-based multiparameter assay to quantify and phenotype SARS-CoV-2 specific T cells medRxiv - the preprint server for health sciences : doi.org/10.1101/2020.10.30.20223099
  20. Riou, C. et al. (2021) Profile of SARS-CoV-2-specific CD4 T cell response: Relationship with disease severity and impact of HIV-1 and active Mycobacterium tuberculosis co-infection medRxiv - the preprint server for health sciences : doi.org/10.1101/2021.02.16.21251838
  21. Willingham, S. B. et al. (2020) Characterization and Phase 1 Trial of a B Cell Activating Anti-CD73 Antibody for the Immunotherapy of COVID-19 medRxiv - the preprint server for health sciences : doi.org/10.1101/2020.09.10.20191486
  22. Li, Z. et al. (2020) SARS-CoV-2-specific T cell memory is long-lasting in the majority of convalsecent COVID-19 individuals bioRxiv - the preprint server for Biology : doi.org/10.1101/2020.11.15.383463
  23. Singh, D. K. et al. (2020) SARS-CoV-2 infection leads to acute infection with dynamic cellular and inflammatory flux in the lung that varies across nonhuman primate species bioRxiv - the preprint server for Biology : doi.org/10.1101/2020.06.05.136481
  24. Thieme, C. J. et al. (2020) Robust T Cell Response Toward Spike, Membrane, and Nucleocapsid SARS-CoV-2 Proteins Is Not Associated with Recovery in Critical COVID-19 Patients. Cell Rep Med. 1(6): 100092
  25. Demaret, J. et al. (2020) Severe SARS‐CoV‐2 patients develop a higher specific T‐cell response Clin Transl Immunology 9(12): e1217
  26. Ferreras, C. et al. (2020) SARS-CoV-2-Specific Memory T Lymphocytes From COVID-19 Convalescent Donors: Identification, Biobanking, and Large-Scale Production for Adoptive Cell Therapy Front Cell Dev Biol. 9: 9:620730
  27. Anft, M. et al. (2021) Detection of pre-existing SARS-CoV-2-reactive T cells in unexposed renal transplant patients J Nephrol. 34(4): 1025-1037
  28. Tamminen, K. et al. (2021) Fusion Protein of Rotavirus VP6 and SARS-CoV-2 Receptor Binding Domain Induces T Cell Responses Vaccines (Basel) 9(7): 733
  29. Ram, R. et al. (2021) Safety and Immunogenicity of the BNT162b2 mRNA COVID-19 Vaccine in Patients after Allogeneic HCT or CD19-based CART therapy-A Single-Center Prospective Cohort Study Transplant Cell Ther. 27(9): 788-749
  30. Vallejo, A. et al.
    IFN-γ
    +
    cell response and IFN-γ release concordance after
    in vitro
    SARS-CoV-2 stimulation
    Eur J Clin Invest (doi: 10.1111/eci.13636)
  31. Aiello, A. et al. (2021) Spike is the most recognized antigen in the whole-blood platform in both acute and convalescent COVID-19 patients Int J Infect Dis 106: 338-347

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