I have done immunopeptidomics for B16F10 melanoma cell line and would like to prioritize my peptides to find an immunogenic target. what is the best in vitro T cell based assay (using the isolated TILs from the murine melanoma tumor) !! any tested protocols !! or suggestions !!
Thank you.I have done Intracellular Cytokine Staining for IFN-g and it worked after co-culturing the TILs with BMDCs stimulated with the target peptides. However, I wonder if someone has established more reliable method such as CFSE proliferation assay !! (Tetramers are expensive).
Hi Mona,
IFN-g ELISpot would be a little closer to funcionality since the T cells have to secrete IFN-g actively. It is cheaper than Tetramers and directly proves not only activation but also recognition and more important functionality. Next step closer to true antitumoral activity would be a funkctional kill assay. Either chromium-release or any form of fluorescence-based cell death detection.
Unkind question: is this still of interest for B16F10???
Good luck anyway,
Michael
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