Rapid Identification of the Tumor-Specific Reactive TIL Repertoire via Combined Detection of CD137, TNF, and IFNγ, Following Recognition of Autologous Tumor-Antigens

• DOI: 10.3389/fimmu.2021.705422
• Publication Date: 2021-10-13T17:09:44Z
• Authors: Arianna Draghi; Christopher Aled ...; Shawez Khan; Krisztian Papp; Martin Lauss; Samuele Soraggi; Haja Dominike Radic; Mario Presti; Katja Harbst; Aishwarya Gokuldass; Anders Kverneland; Morten Nielsen; Marie Christine W...; Mads Hald Andersen; Istvan Csabai; Göran Jönsson; Zoltan Szallasi; Inge Marie Svane; Marco Donia
• Keywords: CD137; Tumor-infiltrating lymphocytes; Tumor Antigen

Detecting the entire repertoire of tumor-specific reactive tumor-infiltrating lymphocytes (TILs) is essential for investigating their immunological functions in tumor microenvironment. Current vitro assays identifying functional activation measure upregulation surface molecules, de novo production antitumor cytokines, or mobilization cytotoxic granules following recognition tumor-antigens, yet there no widely adopted standard method. Here we established an enhanced, simple, method simultaneously CD8+ and CD4+ TILs vitro, using a combination known available flow cytometry assays. By combining detection intracellular CD137 TNF IFNγ after naturally-presented antigens, demonstrate that larger fraction can be detected compared to commonly used This assay revealed multiple polyfunctionality-based clusters both TILs. In situ, combined TNFRSF9, TNF, IFNG identified most TIL repertoire. conclusion, describe straightforward efficient identification which rapidly cancer immunology laboratories.