Identification of Tertiary Lymphoid Structure-Associated Follicular Helper T Cells in Human Tumors and Tissues

Methods Mol Biol. 2018:1845:205-222. doi: 10.1007/978-1-4939-8709-2_12.

Abstract

Follicular helper T (Tfh) cells are major components of the humoral immune response due to their pivotal role in germinal center formation and antibody affinity maturation following B-cell isotype switching. This CD4+ T-cell subtype is mainly found in the B-cell zone of secondary lymphoid organs as well as in tertiary lymphoid structures (TLS), which are highly organized structures composed of T and B cells, occasionally found at the invasive margin in the tumor microenvironment.We describe here how to perform immunofluorescence staining of tumor tissue sections and multicolor flow cytometry on tumor cell suspensions to identify and visualize these TLS-associated Tfh cells within the tumor microenvironment of various human cancers. These assays take advantage of combinations of markers and molecules involved in Tfh differentiation and function.

Keywords: Bcl-6; CXCR5; Flow cytometry; Follicular helper T cell; Immunofluorescence; Multiplex; PD-1; Paraffin-embedded tissue; Tertiary lymphoid structure; Tumor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers
  • Data Analysis
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Humans
  • Immunophenotyping / methods
  • Neoplasms / immunology*
  • Neoplasms / metabolism
  • Neoplasms / pathology*
  • T-Lymphocyte Subsets / immunology
  • T-Lymphocyte Subsets / metabolism
  • T-Lymphocytes, Helper-Inducer / immunology*
  • T-Lymphocytes, Helper-Inducer / metabolism
  • Tertiary Lymphoid Structures / immunology*
  • Tertiary Lymphoid Structures / metabolism
  • Tertiary Lymphoid Structures / pathology*
  • Tumor Microenvironment*

Substances

  • Biomarkers