Human Germinal Center Reaction: Assessing T Follicular Helper-B Cells Interaction In Vitro

Methods Mol Biol. 2021:2285:173-189. doi: 10.1007/978-1-0716-1311-5_15.

Abstract

Antibody responses deeply rely on the interaction of antigen-primed B cells and CD4 helper T cells in the context of germinal center reactions, through signals provided by costimulatory molecules and cytokines. B-cell proliferation and differentiation in antibody-secreting plasma cells are processes that critically depend on the helper function of a specific CD4 T-cell subset, known as follicular helper T cells (Tfh). Here, we describe a method that mimics in vitro the cross talk between Tfh and B cells occurring in the germinal center. The procedure is based on setting up a coculture system with B cells and Tfh isolated from blood of healthy donors, or tonsils removed upon surgical intervention, in order to recapitulate in vitro the Tfh-dependent mechanisms leading to B cells' activation, proliferation, and differentiation.

Keywords: Class switch recombination; Coculture; Follicular helper T cells (Tfh); Germinal center (GC); Naïve B cells; Plasma cells differentiation; Proliferation.

MeSH terms

  • B-Lymphocytes / immunology*
  • B-Lymphocytes / metabolism
  • Cell Communication*
  • Cell Differentiation
  • Cell Proliferation
  • Cell Separation*
  • Cells, Cultured
  • Coculture Techniques
  • Cytokines / metabolism
  • Enzyme-Linked Immunospot Assay
  • Flow Cytometry*
  • Germinal Center / cytology
  • Humans
  • Lymphocyte Activation
  • Palatine Tonsil / cytology
  • Phenotype
  • Research Design
  • T Follicular Helper Cells / immunology*
  • T Follicular Helper Cells / metabolism
  • Workflow

Substances

  • Cytokines