Detection of Tumor Antigen-Specific T-Cell Responses After Oncolytic Vaccination

  • Jonathan G. Pol
  • Byram W. Bridle
  • Brian D. LichtyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2058)


Oncolytic vaccines, which consist of recombinant oncolytic viruses (OV) encoding tumor-associated antigens (TAAs), have demonstrated potent antitumor efficacy in preclinical models and are currently evaluated in phase I/II clinical trials. On one hand, oncolysis of OV-infected malignant entities reinstates cancer immunosurveillance. On the other hand, overexpression of TAAs in infected cells further stimulates the adaptive arm of antitumor immunity. Particularly, the presence of tumor-specific CD8+ T lymphocytes within the tumor microenvironment, as well as in the periphery, has demonstrated prognostic value for cancer treatments. These effector CD8+ T cells can be detected through their production of the prototypical Tc1 cytokine: IFN-γ. The quantitative and qualitative assessment of this immune cell subset remains critical in the development process of efficient cancer vaccines, including oncolytic vaccines. The present chapter will describe a single-cell immunological assay, namely the intracellular cytokine staining (ICS), that allows the enumeration of IFN-γ-producing TAA-specific CD8+ T cells in various tissues (tumor, blood, lymphoid organs) following oncolytic vaccination.

Key words

Oncolytic virus Cancer vaccine Tumor antigen IFN-γ Intracellular cytokine staining (ICS) Flow cytometry 



J.G.P. was supported by the Seerave Foundation. B.W.B. was supported by the Terry Fox Research Institute. B.D.L. was supported by the Terry Fox Foundation, the Ontario Institute for Cancer Research, BioCanRx, and Turnstone Biologics.

Declaration of Interests: J.G.P., B.W.B., and B.D.L. share ownership of patents for cancer vaccination involving oncolytic vaccines. B.D.L. is cofounder and equity holder of and is on the board of directors of Turnstone Biologics.


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Jonathan G. Pol
    • 1
    • 2
    • 3
    • 4
    • 5
  • Byram W. Bridle
    • 6
  • Brian D. Lichty
    • 7
    • 8
    Email author
  1. 1.Gustave Roussy Comprehensive Cancer InstituteVillejuifFrance
  2. 2.INSERM, U1138ParisFrance
  3. 3.Equipe 11 Labellisée par la Ligue Nationale Contre le CancerCentre de Recherche des CordeliersParisFrance
  4. 4.Université de ParisParisFrance
  5. 5.Sorbonne UniversitéParisFrance
  6. 6.Department of Pathobiology, Ontario Veterinary CollegeUniversity of GuelphGuelphCanada
  7. 7.Department of Pathology and Molecular Medicine, McMaster Immunology Research CentreMcMaster UniversityHamiltonCanada
  8. 8.Turnstone BiologicsOttawaCanada

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