STAR Protocols (Jun 2022)

Flow cytometry analysis of endothelial cells and subsets of exhausted CD8+ T cells in murine tumor models

  • Lucas Blanchard,
  • Estefania Vina,
  • Assia Asrir,
  • Claire Tardiveau,
  • Juliette Coudert,
  • Robin Laffont,
  • Dorian Tarroux,
  • Sarah Bettini,
  • Krystle Veerman,
  • Fanny Lafouresse,
  • Mélanie Pichery,
  • Emilie Mirey,
  • Elisabeth Bellard,
  • Nathalie Ortega,
  • Jean-Philippe Girard

Journal volume & issue
Vol. 3, no. 2
p. 101444

Abstract

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Summary: Here, we present a protocol for flow cytometry analysis of endothelial cells (ECs) and CD8+ T cells in murine tumor models, at baseline and after cancer immunotherapy with anti-PD-1/anti-CTLA-4 antibodies. We provide gating strategies for identification of specific cell subsets including ECs from tumor-associated high endothelial venules (TA-HEVs), stem-like, and terminally exhausted CD8+ T cells. This protocol represents a valuable tool for the analysis of rare subsets of tumor ECs and CD8+ T cells with critical roles in antitumor immunity.For complete details on the use and execution of this protocol, please refer to Asrir et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords