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Updated: Jun 27, 2026

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
08:26

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

Published on: December 14, 2016

Peripheral blood CD27+ IgG+ B cells rapidly proliferate and differentiate into immunoglobulin-secreting cells after

Jessie F Fecteau1, Annie Roy, Sonia Néron

  • 1Héma-Québec, Ingénierie cellulaire, Recherche et développement, Québec, Canada.

Immunology
|November 20, 2008
PubMed

Insights

Human memory B cells, particularly IgG(+) types, show rapid differentiation and high immunoglobulin secretion upon CD40 stimulation. This study reconciles blood and lymphoid memory B cell responses, highlighting circulating memory IgG(+) cells

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • In vitro CD40 stimulation of human B cells from lymphoid organs shows memory B cells proliferate and differentiate faster than naive B cells.
  • Previous studies indicated blood memory B cells primarily differentiate into immunoglobulin-secreting cells post-CD40 stimulation.
  • Variations in CD40-CD154 interaction are known to impact B cell fate.

Purpose of the Study:

  • To compare the in vitro response of distinct blood B cell subsets (CD27(-) and CD27(+) IgG(+)) to CD154 stimulation.
  • To monitor B cell population dynamics during short-term and long-term CD154 exposure.
  • To reconcile differences in B cell responses between lymphoid organs and blood.

Main Methods:

  • Isolation and in vitro stimulation of human peripheral blood B cells (CD27(-) and CD27(+) IgG(+)).
  • Exposure to CD154 in the presence of IL-2, IL-4, and IL-10.
  • Monitoring of B cell population evolution over 5 days and during long-term stimulation using flow cytometry (CD19, CD27, CD38, IgG).

Main Results:

  • CD27(+) B cells differentiated more readily into immunoglobulin-secreting cells than CD27(-) cells within 5 days.
  • CD27(+) IgG(+) B cells generated a homogeneous population of CD19(+) CD27(++) CD38(+) IgG(lo) cells with high IgG secretion.
  • CD27(-) IgG(-) B cells partially converted to CD19(++) CD27(-) CD38(-) IgG(++) cells but did not secrete IgG.
  • Long-term stimulation showed CD27(+) IgG(+) B cells maintained high expansion and differentiation capacity over naive B cells.
  • Long-term stimulation induced naive homeostatic proliferation in CD27(-) IgG(-) and total CD19(+) B cells.

Conclusions:

  • Circulating memory IgG(+) B cells are intrinsically prone to rapid activation and differentiation upon appropriate stimulation.
  • The study reconciles memory B cell responses from blood and lymphoid organs, showing preferential differentiation capacity compared to naive cells.
  • Distinct subsets of memory B cells exhibit unique responses to CD40-CD154 signaling, influencing their differentiation pathways.

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