Modulation of single-cell IgG secretion frequency and rates in human memory B cells by CpG DNA, CD40L, IL-21, and cell division

J Immunol. 2009 Sep 1;183(5):3177-87. doi: 10.4049/jimmunol.0804233. Epub 2009 Aug 12.

Abstract

During the recall response by CD27(+) IgG class-switched human memory B cells, total IgG secreted is a function of the following: 1) the number of IgG-secreting cells (IgG-SC), and 2) the secretion rate of each cell. In this study, we report the quantitative ELISPOT method for simultaneous estimation of single-cell IgG secretion rates and secreting cell frequencies in human B cell populations. We found that CD27(+) IgM(-) memory B cells activated with CpG and cytokines had considerable heterogeneity in the IgG secretion rates, with two major secretion rate subpopulations. BCR cross-linking reduced the frequency of cells with high per-cell IgG secretion rates, with a parallel decrease in CD27(high) B cell blasts. Increased cell death may account for the BCR-stimulated reduction in high-rate IgG-SC CD27(high) B cell blasts. In contrast, the addition of IL-21 to CD40L plus IL-4-activated human memory B cells induced a high-rate IgG-SC population in B cells with otherwise low per-cell IgG secretion rates. The profiles of human B cell IgG secretion rates followed the same biphasic distribution and range irrespective of division class. This, along with the presence of non-IgG-producing, dividing B cells in CpG plus cytokine-activated B memory B cell populations, is suggestive of an on/off switch regulating IgG secretion. Finally, these data support a mixture model of IgG secretion in which IgG secreted over time is modulated by the frequency of IgG-SC and the distribution of their IgG secretion rates.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • B-Lymphocyte Subsets / immunology*
  • B-Lymphocyte Subsets / metabolism*
  • B-Lymphocyte Subsets / pathology
  • CD40 Ligand / physiology*
  • Cell Death / immunology
  • Cell Division / immunology*
  • Cell Line, Tumor
  • Cell Proliferation
  • Cells, Cultured
  • CpG Islands / immunology*
  • Cytokines / physiology
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoglobulin G / metabolism*
  • Immunologic Memory*
  • Interleukins / physiology*
  • Lymphocyte Activation / immunology
  • Mice
  • NIH 3T3 Cells
  • Oligodeoxyribonucleotides / pharmacology*

Substances

  • CPG-oligonucleotide
  • Cytokines
  • Immunoglobulin G
  • Interleukins
  • Oligodeoxyribonucleotides
  • CD40 Ligand
  • interleukin-21