Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

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

Draghi, Arianna ; Chamberlain, Christopher Aled ; Khan, Shawez ; Papp, Krisztian ; Lauss, Martin LU ; Soraggi, Samuele ; Radic, Haja Dominike ; Presti, Mario ; Harbst, Katja LU orcid and Gokuldass, Aishwarya , et al. (2021) In Frontiers in Immunology 12.
Abstract

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... (More)

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.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; ; and , et al. (More)
; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; and (Less)
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
CD137 (4-1BB), immune-monitoring, immune-responses to cancer, single-cell technologies, tumor-infiltrating lymphocytes (TILs), tumor-specific activation, tumor-specific reactivity
in
Frontiers in Immunology
volume
12
article number
705422
publisher
Frontiers Media S. A.
external identifiers
  • pmid:34707600
  • scopus:85117920197
ISSN
1664-3224
DOI
10.3389/fimmu.2021.705422
language
English
LU publication?
yes
additional info
Publisher Copyright: © Copyright © 2021 Draghi, Chamberlain, Khan, Papp, Lauss, Soraggi, Radic, Presti, Harbst, Gokuldass, Kverneland, Nielsen, Westergaard, Andersen, Csabai, Jönsson, Szallasi, Svane and Donia.
id
183dcc6c-c7cb-4ab8-9312-203a9d107a96
date added to LUP
2021-11-23 15:37:45
date last changed
2024-06-17 00:06:40
@article{183dcc6c-c7cb-4ab8-9312-203a9d107a96,
  abstract     = {{<p>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<sup>+</sup> and CD4<sup>+</sup> 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<sup>+</sup> TILs can be detected in vitro compared to commonly used assays. This assay revealed multiple polyfunctionality-based clusters of both CD4<sup>+</sup> and CD8<sup>+</sup> 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.</p>}},
  author       = {{Draghi, Arianna and Chamberlain, Christopher Aled and Khan, Shawez and Papp, Krisztian and Lauss, Martin and Soraggi, Samuele and Radic, Haja Dominike and Presti, Mario and Harbst, Katja and Gokuldass, Aishwarya and Kverneland, Anders and Nielsen, Morten and Westergaard, Marie Christine Wulff and Andersen, Mads Hald and Csabai, Istvan and Jönsson, Göran and Szallasi, Zoltan and Svane, Inge Marie and Donia, Marco}},
  issn         = {{1664-3224}},
  keywords     = {{CD137 (4-1BB); immune-monitoring; immune-responses to cancer; single-cell technologies; tumor-infiltrating lymphocytes (TILs); tumor-specific activation; tumor-specific reactivity}},
  language     = {{eng}},
  month        = {{10}},
  publisher    = {{Frontiers Media S. A.}},
  series       = {{Frontiers in Immunology}},
  title        = {{Rapid Identification of the Tumor-Specific Reactive TIL Repertoire via Combined Detection of CD137, TNF, and IFNγ, Following Recognition of Autologous Tumor-Antigens}},
  url          = {{http://dx.doi.org/10.3389/fimmu.2021.705422}},
  doi          = {{10.3389/fimmu.2021.705422}},
  volume       = {{12}},
  year         = {{2021}},
}