Enzyme-linked ImmunoSpot (ELISpot) assay to quantify peptide-specific IFN-γ production by splenocytes in a mouse tumor model after radiation therapy.

Fiche publication

Date publication

septembre 2024

Journal

Methods in cell biology

Auteurs

Membres identifiés du Cancéropôle Est :
Pr ADOTEVI Olivier , Dr BOUSTANI Jihane

Tous les auteurs :
Lecoester B, Xie Y, Marguier A, Boulerot L, Malfroy M, Adotévi O, Boustani J

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/39393885

Résumé

To develop new effective therapeutic strategies for cancer patients, there is a need for extensive and precise insights into the mechanisms involved in the immune response to anti-cancer treatments. The enzyme-linked immunospot (ELISpot) assay is a rapid and reproducible technique that allows the detection of cytokine-producing antigen-specific T cells at the single cell level. This protocol describes an interferon gamma (IFN-γ) ELISpot method for measuring antigen-specific murine CD8 T cells that produce IFN-γ, a marker of their activation and cytotoxicity. Splenocytes from tumor-bearing mice treated with radiation therapy were used as source of CD8 T cells and were stimulated with a tumor-derived peptide. This method was facilitated by a ready-to-use assay kit and provides a tool to analyze the specificity, intensity, and kinetics of specific CD8 T cells.