Cytokine profiling of tumor-infiltrating T lymphocytes by flow cytometry

Methods Enzymol. 2020:631:1-20. doi: 10.1016/bs.mie.2019.08.015. Epub 2019 Oct 18.

Abstract

Intracellular cytokine staining (ICS) utilizing fluorescently labeled, cytokine-specific antibodies is a powerful technique utilized to evaluate cytokine expression that provides resolution at the single cell level. Combined with multi-parameter flow cytometry, ICS can provide detailed information on complex cytokine profiles and cellular phenotypes within the tumor microenvironment, particularly for the CD4+ T helper and CD8+ cytotoxic T cell compartments. This technique provides critical information concerning tumor-infiltrating T cell function that is essential for evaluating existing or therapeutically-induced tumor antigen-specific T cell responses in both preclinical models and cancer patients. In this chapter we will discuss in detail the critical steps necessary for a successful ICS assay and outline common protocols for the evaluation of cytokine production from T cell subsets present within the tumor microenvironment.

Keywords: Cancer; Intracellular cytokine staining; Tumor-infiltrating lymphocytes.

MeSH terms

  • Animals
  • Cytokines / analysis*
  • Flow Cytometry / methods*
  • Humans
  • Immunologic Techniques / methods*
  • Lymphocytes, Tumor-Infiltrating / metabolism*
  • T-Lymphocytes / metabolism
  • Tumor Microenvironment / immunology*

Substances

  • Cytokines