Kinetics of B cell receptor signaling in human B cell subsets mapped by phosphospecific flow cytometry

J Immunol. 2006 Aug 1;177(3):1581-9. doi: 10.4049/jimmunol.177.3.1581.

Abstract

Differences in BCR signaling may govern outcomes as diverse as proliferation and cell death. We profiled BCR signaling kinetics in subsets of primary human B cells using flow cytometry. In the predominant population expressing IgM, BCR cross-linking led to a quick burst of Syk, ERK1/2, and p38 signaling. In contrast, IgG B cells sustained higher per-cell ERK1/2 phosphorylation over time. This dichotomy suggested a mechanism for dampening signals transmitted by IgM. Regulatory phosphatase activity in IgM B cells was BCR-mediated and initiated more slowly than kinase activity. This BCR-mediated phosphatase activity was sensitive to inhibition by H(2)O(2) and required to attenuate IgM BCR signaling. These results provide the first kinetic maps of BCR signaling in primary human B cell subsets and enable new studies of signaling in B cell disorders, such as autoimmunity and cancer.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agammaglobulinaemia Tyrosine Kinase
  • B-Lymphocyte Subsets / enzymology
  • B-Lymphocyte Subsets / immunology*
  • B-Lymphocyte Subsets / metabolism*
  • Flow Cytometry / methods*
  • Humans
  • Hydrogen Peroxide / chemistry
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / metabolism
  • Immunoglobulin M / biosynthesis
  • Immunoglobulin M / metabolism
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Kinetics
  • MAP Kinase Signaling System / immunology*
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 1 / physiology
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Mitogen-Activated Protein Kinase 3 / physiology
  • Peptide Mapping / methods
  • Phosphorylation
  • Protein-Tyrosine Kinases / metabolism
  • Receptors, Antigen, B-Cell / metabolism
  • Receptors, Antigen, B-Cell / physiology*
  • Syk Kinase

Substances

  • Immunoglobulin G
  • Immunoglobulin M
  • Intracellular Signaling Peptides and Proteins
  • Receptors, Antigen, B-Cell
  • Hydrogen Peroxide
  • Protein-Tyrosine Kinases
  • Agammaglobulinaemia Tyrosine Kinase
  • SYK protein, human
  • Syk Kinase
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3