An in vitro model of differentiation of memory B cells into plasmablasts and plasma cells including detailed phenotypic and molecular characterization

Jourdan, Michel; Caraux, Anouk; Vos, John De; Fiol, Geneviève; Larroque, Marion; Cognot, Chantal; Bret, Caroline; Duperray, Christophe; Hose, Dirk; Klein, Bernard

doi:10.1182/blood-2009-07-235960

Cited by 208 publications

(299 citation statements)

References 40 publications

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“…B-cell purity was always higher than 95% as assessed by flow cytometry. Purified B cells were cultured in complete Iscove modified Dulbecco medium (IMDM) and stimulated for 10 days with crosslinked CD40L, CpG-B, IL-2, IL-6, IL-10, IL-15, and interferon-a (IFN-a) in a 3-step sequential differentiation method as described by Jourdan et al 41 At days 1, 4, 7, and 10, B-cell phenotype on freshly harvested cells, and IL-10 production by B cells after 5-hour PMA 1 ionomycin in vitro stimulation, were assessed by flow cytometry as described in "Cell culture and B-cell IL-10 production. "…”

Section: In Vitro Plasmablast Differentiationmentioning

confidence: 99%

CD24hiCD27+ and plasmablast-like regulatory B cells in human chronic graft-versus-host disease

Masson

Bouaziz

Buanec

et al. 2015

Blood

140

119

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Key Points• Chronic graft-versus-host disease is associated with a global Breg defect.• This defect is particularly accentuated in the CD24 hi CD27 1 Breg compartment.Interleukin 10 (IL-10)-producing B cells (regulatory B cells [Bregs]) regulate autoimmunity in mice and humans, and a regulatory role of IL-10-producing plasma cells has been described in mice. Dysfunction of B cells that maintain homeostasis may play a role in the pathogenesis of chronic graft-versus-host disease (cGVHD) after allogeneic stem cell transplantation. Here, we found a relation between decreased Breg frequencies and cGVHD severity. An impaired ability of B cells to produce IL-10, possibly linked to poor signal transducer and activator of transcription 3 and extracellular signal-regulated kinase phosphorylation, was found in patients with active cGVHD. IL-10 production was not confined to a single B-cell subset, but enriched in both the CD24 hi CD27 1 and CD27 hi CD38 hi plasmablast B-cell compartments. In vitro plasmablast differentiation increased the frequency of IL-10-producing B cells. We confirmed that allogeneic transplant recipients had an impaired reconstitution of the memory B-cell pool. cGVHD patients had less CD24 hi CD27 1 B cells and IL-10-producing CD24 hi CD27 1 B cells. Patients with cGVHD had increased plasmablast frequencies but decreased IL-10-producing plasmablasts. These results suggest a role of CD24 hi CD27 1 B-cell and plasmablast-derived IL-10 in the regulation of human cGVHD. (Blood. 2015;125(11):1830-1839 IntroductionChronic graft-versus-host disease (cGVHD) is the leading cause of morbi-mortality after allogeneic hematopoietic stem cell transplantation (AHSCT). 1 GVHD prevention by means of adoptive transfer of regulatory T cells (Tregs) 2,3 and cGVHD treatment by in vivo induction of Tregs by low-dose interleukin 2 (IL-2) 4,5 may be effective. The exact role of IL-10-producing regulatory B cells (Bregs) in cGVHD is yet to be understood. Bregs have been shown to downmodulate adaptive 6 or innate immune responses 7 in mice and humans. In cGVHD, B cells are essentially recognized as positive regulators of inflammation. 1,8,9 Increased B-cell receptor responsiveness was found in cGVHD patients. 10 B-cell homeostatic defects were described in cGVHD, including elevated B-cell activating factor of the tumor necrosis factor (TNF) family (BAFF)/B-cell ratios, 11-15 expansion of CD21 lo B cells, 16 reduced CD5 1 B1-like cell numbers, 17 decreased CD27 1 memory B cells, and hypogammaglobulinemia. 18 BAFF concentrations correlated with increased circulating pre-germinal center (GC) B-cell and post-GC plasmablast cell counts in patients with cGVHD. 11 B-cell depletion with rituximab prevented cGVHD in humans. 19 Patients with cGVHD frequently have circulating antibodies reactive to recipient cells. 1,9,20 Besides their functions in antibody secretion, cytokine and chemokine production, and antigen presentation, B cells exhibit regulatory properties in several human autoimmune diseases including systemic lupus erythemat...

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Section: In Vitro Plasmablast Differentiationmentioning

confidence: 99%

CD24hiCD27+ and plasmablast-like regulatory B cells in human chronic graft-versus-host disease

Masson

Bouaziz

Buanec

et al. 2015

Blood

140

119

View full text Add to dashboard Cite

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“…plasmablasts as well as with non-cell cycling PCs, 19 whereas H/F-LVs efficiently transduce both plasmablasts and PCs. Thus, H/F-LVs provide a powerful tool to stably introduce and express coding DNA in healthy or primary malignant PCs.…”

Section: Discussionmentioning

confidence: 99%

“…33 The combination of the ability to generate human PCs in vitro and to efficiently induce the expression of a given gene in these cells should make the study of healthy human PCs far easier. 19 An interesting application will be to immortalize human PCs to produce human monoclonal antibodies specific to a given antigen, introducing the major genes deregulated in malignant PCs and yielding to HMCLs growing in vitro. In conclusion, the finding that LVs pseudotyped with MV envelope glycoproteins enable an efficient and stable introduction and expression of a given gene in noncycling normal and malignant PCs is a major breakthrough, that will further stimulate the study and understanding of the most important mechanisms controlling normal and malignant PC biology.…”

Section: Discussionmentioning

confidence: 99%

“…16 --18 Healthy plasmablasts or PCs were obtained using our three-step in vitro model making it possible to obtain six syndecan-1 þ PCs starting from one circulating memory B cell within 10 days. 19 This threestep model mimics the activation and differentiation processes of B cells mediated at least by T cells and follicular dendritic cells occurring in germinal centers.…”

Section: Cell Samplesmentioning

confidence: 99%

“…21 Transduction of healthy and malignant PCs Cells were cultured in RPMI 1640 medium (Invitrogen, Gibco BRL, Cergy Pontoise, France) supplemented with 10% FCS (PAA Laboratory GmbH, Pasching, Austria) and with 2 ng/ml of IL-6 (Abcys SA, Paris, France) for malignant PCs and HMCLs and in the appropriate step culture conditions for healthy in vitro generated PCs. 19 A 2 Â 10 5 cells were seeded per 48-well and were transduced with a multiplicity of infection ranging from 0.125 to 1.5 for HMCLs and up to 7.5 as indicated for malignant or healthy PCs. The percentage of GFP þ cells was determined 72 h after transduction by fluorescence-activated cell sorting (FACS) using a FACScan cytometer (Becton Dickinson (BD), San Jose, CA, USA).…”

Section: Lentiviral Productionmentioning

confidence: 99%

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Efficient transduction of healthy and malignant plasma cells by lentiviral vectors pseudotyped with measles virus glycoproteins

et al. 2012

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Autoreactive Plasmablasts After B Cell Depletion With Rituximab and Relapses in Antineutrophil Cytoplasmic Antibody–Associated Vasculitis

Berti

Hillion

Konig

et al. 2023

Arthritis & Rheumatology

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Objective. Autoreactive B cells are responsible for antineutrophil cytoplasmic antibody (ANCA) production in ANCA-associated vasculitis (AAV). Rituximab (RTX) depletes circulating B cells, including autoreactive B cells. We aimed to evaluate changes and associations with relapse of the circulating autoreactive B cell pool following therapeutic B cell depletion in AAV.Methods. Sequential flow cytometry was performed on 148 samples of peripheral blood mononuclear cells from 23 patients with proteinase 3 (PR3)-ANCA-positive AAV who were treated with RTX for remission induction and monitored after stopping therapy during long-term follow-up in a prospective clinical trial. PR3 was used as a ligand to target autoreactive PR3-specific (PR3+) B cells. B cell recurrence was considered as the first blood sample with ≥10 B cells/μl after RTX treatment.Results. At B cell recurrence, PR3+ B cell frequency among B cells was higher than baseline (P < 0.01). Within both PR3+ and total B cells, frequencies of transitional and naive subsets were higher at B cell recurrence than at baseline, while memory subsets were lower (P < 0.001 for all comparisons). At B cell recurrence, frequencies of B cells and subsets did not differ between patients who experienced relapse and patients who remained in remission. In contrast, the plasmablast frequency within the PR3+ B cell pool was higher in patients who experienced relapse and associated with a shorter time to relapse. Frequencies of PR3+ plasmablasts higher than baseline were more likely to be found in patients who experienced relapse within the following 12 months compared to those in sustained remission (P < 0.05).Conclusion. The composition of the autoreactive B cell pool varies significantly following RTX treatment in AAV, and early plasmablast enrichment within the autoreactive pool is associated with future relapses.

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An in vitro model of differentiation of memory B cells into plasmablasts and plasma cells including detailed phenotypic and molecular characterization

Cited by 208 publications

References 40 publications

CD24hiCD27+ and plasmablast-like regulatory B cells in human chronic graft-versus-host disease

CD24hiCD27+ and plasmablast-like regulatory B cells in human chronic graft-versus-host disease

Efficient transduction of healthy and malignant plasma cells by lentiviral vectors pseudotyped with measles virus glycoproteins

Autoreactive Plasmablasts After B Cell Depletion With Rituximab and Relapses in Antineutrophil Cytoplasmic Antibody–Associated Vasculitis

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