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Updated: Jan 20, 2026
In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model
Published on: November 28, 2017
Detection of Tumor Antigen-Specific T-Cell Responses After Oncolytic Vaccination
Jonathan G Pol1,2,3,4,5, Byram W Bridle6, Brian D Lichty7,8
1Gustave Roussy Comprehensive Cancer Institute, Villejuif, France. pol_jonathan@yahoo.fr.
Abstract:
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.
Insights
Oncolytic vaccines harness oncolytic viruses (OV) to stimulate antitumor immunity. This study details an assay to measure TAA-specific CD8+ T cells, crucial for evaluating vaccine effectiveness.
Area of Science:
- Immunology
- Oncology
- Vaccinology
Background:
- Oncolytic vaccines (OV) encode tumor-associated antigens (TAAs) to enhance antitumor responses.
- OV induce cancer cell lysis and TAA overexpression, stimulating adaptive immunity.
- Tumor-specific CD8+ T cells, particularly IFN-γ producers, are critical for prognosis and treatment efficacy.
Purpose of the Study:
- To describe an immunological assay for quantifying TAA-specific CD8+ T cells.
- To enable assessment of immune responses following oncolytic vaccination.
- To support the development of effective cancer vaccines.
Main Methods:
- Intracellular Cytokine Staining (ICS) assay described.
- Enumeration of IFN-γ-producing TAA-specific CD8+ T cells.
- Analysis performed on various tissues including tumor, blood, and lymphoid organs.
Main Results:
- The ICS assay allows for the detection of specific immune cell subsets.
- Quantitative and qualitative assessment of TAA-specific CD8+ T cells is achievable.
- The assay is applicable to samples from different anatomical sites.
Conclusions:
- Intracellular Cytokine Staining (ICS) is a valuable method for evaluating oncolytic vaccines.
- Measuring IFN-γ-producing CD8+ T cells provides critical insights into vaccine-induced immunity.
- This assay aids in the development and optimization of cancer immunotherapies.
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