Rapid Identification of the Tumor-Specific Reactive TIL Repertoire via Combined Detection of CD137, TNF, and IFNϒ, Following Recognition of Autologous Tumor-Antigens
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Detecting the entire repertoire of tumor-specific reactive tumor-infiltrating lymphocytes (TILs) is essential for investigating their immunological functions in the tumor microenvironment. Current in vitro assays identifying tumor-specific functional activation measure the upregulation of surface molecules, de novo production of antitumor cytokines, or mobilization of cytotoxic granules following recognition of tumor-antigens, yet there is no widely adopted standard method. Here we established an enhanced, yet simple, method for identifying simultaneously CD8+ and CD4+ tumor-specific reactive TILs in vitro, using a combination of widely known and available flow cytometry assays. By combining the detection of intracellular CD137 and de novo production of TNF and IFNγ after recognition of naturally-presented tumor antigens, we demonstrate that a larger fraction of tumor-specific and reactive CD8+ TILs can be detected in vitro compared to commonly used assays. This assay revealed multiple polyfunctionality-based clusters of both CD4+ and CD8+ tumor-specific reactive TILs. In situ, the combined detection of TNFRSF9, TNF, and IFNG identified most of the tumor-specific reactive TIL repertoire. In conclusion, we describe a straightforward method for efficient identification of the tumor-specific reactive TIL repertoire in vitro, which can be rapidly adopted in most cancer immunology laboratories.
Originalsprog | Engelsk |
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Artikelnummer | 705422 |
Tidsskrift | Frontiers in Immunology |
Vol/bind | 12 |
Antal sider | 13 |
ISSN | 1664-3224 |
DOI | |
Status | Udgivet - 2021 |
Bibliografisk note
Funding Information:
This work was supported by the by The Danish Cancer Society under Grants R180-A11339, R184-A11806, and R204-A12535; the Lundbeck Foundation under Grants R233-2016-3728, R286-2018-991, and R307-2018-3636; the Independent Research Fund Denmark under Grant 8045-00067B; the Danish National Board of Health under Grant 4-1612-236/8 “Empowering Cancer Immunotherapy in Denmark”; the National Research, Development and Innovation Fund of Hungary under Grant Project no. FIEK_16-1-2016-0005; the Herlev and Gentofte Hospital Research Council under Clinician-Scientist Grant.
Publisher Copyright:
© Copyright © 2021 Draghi, Chamberlain, Khan, Papp, Lauss, Soraggi, Radic, Presti, Harbst, Gokuldass, Kverneland, Nielsen, Westergaard, Andersen, Csabai, Jönsson, Szallasi, Svane and Donia.
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