Analysis by Flow Cytometry of B-Cell Activation and Antibody Responses Induced by Toll-Like Receptors

Methods Mol Biol. 2016:1390:229-48. doi: 10.1007/978-1-4939-3335-8_15.

Abstract

Toll-like receptors (TLRs) are expressed in B lymphocytes and contribute to B-cell activation, antibody responses, and their maturation. TLR stimulation of mouse B cells induces class switch DNA recombination (CSR) to isotypes specified by cytokines, and also induces formation of IgM(+) as well as class-switched plasma cells. B-cell receptor (BCR) signaling, while on its own inducing limited B-cell proliferation and no CSR, can enhance CSR driven by TLRs. Particular synergistic or antagonistic interactions among TLR pathways, BCR, and cytokine signaling can have important consequences for B-cell activation, CSR, and plasma cell formation. This chapter outlines protocols for the induction and analysis of B-cell activation and antibody production by TLRs with or without other stimuli.

Keywords: AID; Antibody; B cells; B-cell receptor (BCR); Class switch DNA recombination (CSR); CpG; Cytokine; Germinal center; Immunoglobulin; Lipopolysaccharides (LPS); T-independent antibody response; Toll-like Receptor (TLR).

MeSH terms

  • Animals
  • Antibody Formation / immunology*
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / metabolism*
  • Biomarkers
  • Flow Cytometry* / methods
  • Immunomagnetic Separation
  • Immunophenotyping
  • Lymphocyte Activation / immunology*
  • Mice
  • Toll-Like Receptors / metabolism*

Substances

  • Biomarkers
  • Toll-Like Receptors