title: Modulation of single-cell IgG secretion frequency and rates in human memory B cells by CpG DNA, CD40L, IL-21, and cell division.
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.
authors.affiliation: Division of Nephrology, University of Rochester Medical Center, Rochester, NY 14642, USA.
authors.forename: Alicia D
authors.initials: AD
authors.lastname: Henn
authors.forename: Jonathan
authors.initials: J
authors.lastname: Rebhahn
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Brown
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Murphy
authors.forename: Mircea N
authors.initials: MN
authors.lastname: Coca
authors.forename: Ollivier
authors.initials: O
authors.lastname: Hyrien
authors.forename: Tina
authors.initials: T
authors.lastname: Pellegrin
authors.forename: Tim
authors.initials: T
authors.lastname: Mosmann
authors.forename: Martin S
authors.initials: MS
authors.lastname: Zand
mesh_terms: D000818:Animals; D016175:B-Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism* / Q000473:pathology; D023201:CD40 Ligand / Q000502:physiology*; D016923:Cell Death / Q000276:immunology; D002455:Cell Division / Q000276:immunology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002478:Cells, Cultured; D018899:CpG Islands / Q000276:immunology*; D016207:Cytokines / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D007156:Immunologic Memory; D007378:Interleukins / Q000502:physiology*; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D041681:NIH 3T3 Cells; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D000098546:Interleukin-21
chemical_list: C408982:CPG-oligonucleotide; D016207:Cytokines; D007074:Immunoglobulin G; D007378:Interleukins; D009838:Oligodeoxyribonucleotides; D023201:CD40 Ligand; D000098546:Interleukin-21
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