Mast cells enhance proliferation of B lymphocytes and drive their differentiation toward IgA-secreting plasma cells

Abstract The evidence of a tight spatial interaction between mast cells (MCs) and B lymphocytes in secondary lymphoid organs, along with the data regarding the abundance of MCs in several B-cell lymphoproliferative disorders prompted us to investigate whether MCs could affect the proliferation and differentiation of B cells. To this aim, we performed coculture assays using mouse splenic B cells and bone marrow–derived MCs. Both nonsensitized and activated MCs proved able to induce a significant inhibition of cell death and an increase in proliferation of naive B cells. Such proliferation was further enhanced in activated B cells. This effect relied on cell-cell contact and MC-derived interleukin-6 (IL-6). Activated MCs could regulate CD40 surface expression on unstimulated B cells and the interaction between CD40 with CD40 ligand (CD40L) on MCs, together with MC-derived cytokines, was involved in the differentiation of B cells into CD138+ plasma cells and in selective immunoglobulin A (IgA) secretion. These data were corroborated by in vivo evidence of infiltrating MCs in close contact with IgA-expressing plasma cells within inflamed tissues. In conclusion, we reported here a novel role for MCs in sustaining B-cell expansion and driving the development of IgA-oriented humoral immune responses.

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B lymphocytes express and lose syndecan at specific stages of differentiation.

Cell Regulation ◽

10.1091/mbc.1.1.27 ◽

1989 ◽

Vol 1 (1) ◽

pp. 27-35 ◽

Cited By ~ 272

Author(s):

R D Sanderson ◽

P Lalor ◽

M Bernfield

Keyword(s):

Bone Marrow ◽

B Cells ◽

B Cell ◽

B Lymphocytes ◽

B Lymphocyte ◽

Plasma Cells ◽

Receptor Expression ◽

Cell Stage ◽

Precursor B Cells

Lymphopoietic cells require interactions with bone marrow stroma for normal maturation and show changes in adhesion to matrix during their differentiation. Syndecan, a heparan sulfate-rich integral membrane proteoglycan, functions as a matrix receptor by binding cells to interstitial collagens, fibronectin, and thrombospondin. Therefore, we asked whether syndecan was present on the surface of lymphopoietic cells. In bone marrow, we find syndecan only on precursor B cells. Expression changes with pre-B cell maturation in the marrow and with B-lymphocyte differentiation to plasma cells in interstitial matrices. Syndecan on B cell precursors is more heterogeneous and slightly larger than on plasma cells. Syndecan 1) is lost immediately before maturation and release of B lymphocytes into the circulation, 2) is absent on circulating and peripheral B lymphocytes, and 3) is reexpressed upon their differentiation into immobilized plasma cells. Thus, syndecan is expressed only when and where B lymphocytes associate with extracellular matrix. These results indicate that B cells differentiating in vivo alter their matrix receptor expression and suggest a role for syndecan in B cell stage-specific adhesion.

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Differential Regulation of Mouse B Cell Development by Transforming Growth Factor β1

Developmental Immunology ◽

10.1080/1044667031000088057 ◽

2002 ◽

Vol 9 (2) ◽

pp. 86-95 ◽

Cited By ~ 8

Author(s):

Denise A. Kaminski ◽

John J. Letterio ◽

Peter D. Burrows

Keyword(s):

B Cells ◽

Growth Factor ◽

B Cell ◽

Transforming Growth Factor ◽

Plasma Cells ◽

Population Analysis ◽

Cell Development ◽

B Cell Development

Transforming growth factor β (TGFβ) can inhibit thein vitroproliferation, survival and differentiation of B cell progenitors, mature B lymphocytes and plasma cells. Here we demonstrate unexpected, age-dependent reductions in the bone marrow (BM) B cell progenitors and immature B cells in TGFβ1-/-mice. To evaluate TGFβ responsiveness during normal B lineage development, cells were cultured in interleukin 7 (IL7)±TGFβ. Picomolar doses of TGFβ1 reduced pro-B cell recoveries at every timepoint. By contrast, the pre-B cells were initially reduced in number, but subsequently increased compared to IL7 alone, resulting in a 4-fold increase in the growth rate for the pre-B cell population. Analysis of purified BM sub-populations indicated that pro-B cells and the earliest BP1-pre-B cells were sensitive to the inhibitory effects of TGFβ1. However, the large BP1+pre-B cells, although initially reduced, were increased in number at days 5 and 7 of culture. These results indicate that TGFβ1 is important for normal B cell developmentin vivo, and that B cell progenitors are differentially affected by the cytokine according to their stage of differentiation.

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Protein Signature of LMP1 Signaling in PTLDs Identifies and Mimics Inter/Perifollicular CD30+ EBV− B Blasts.

Blood ◽

10.1182/blood.v104.11.3251.3251 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3251-3251

Author(s):

Rita Shaknovich ◽

Katia Basso ◽

Govind Bhagat ◽

Bachir Alobeid ◽

Giorgio Cattoretti

Keyword(s):

B Cells ◽

B Cell ◽

Plasma Cells ◽

Cell Subset ◽

Cellular Transformation ◽

Cell Types ◽

Marginal Zone B Cells ◽

Latently Infected ◽

Protein Signature

Abstract EBV-associated B-cell Post-Transpant Lymphoproliferative Disorders (PTLDs) represent a diverse group of lesions morphologically, in clinical presentation and behaviour, ranging from early reversible lesions to monomorphic aggressive lymphomas. Polymorphic cases, which represent the focus of our analysis, contain a mixture of cells in various EBV latency stages, defined by EBNA1, EBNA2 and LMP1 immunostaining. LMP1 is a key viral protein for cellular transformation and, analogously to CD40, engages TNF Receptor Associated Proteins and activates NF-kB and NF-kB-responsive genes. We analyzed the protein signature of LMP1 in PTLDs and non-PTLD tonsils by double staining for LMP1, CD30, CD20, Pax5 and signaling molecules. A remarkably conserved set of proteins, associated with LMP1/CD40 signaling and NF-kB activation is expressed both in the EBV-infected lymphoid population in polymorphic PTLDs and in a normal B-cell subset(s) in reactive tonsils. These proteins include highly expressed CD30, JunB, nuclear cRel, TRAF-1, Bcl-XL, MUM1, CCL22 and downregulated BCL6 and CD10. We observed that EBV infection, possibly through LMP1 and LMP2A signaling, results in varioius degrees of differentiation within the neoplastic clone. EBER+ terminally differentiated mucosa-associated IRTA-1+ marginal zone B-cells and CD138+ plasma cells were identified in most cases, including control post-transplant tonsils with no overt disease. We document for the first time in situ, in-vivo evidence of EBV latently infected post-Germinal Center B cells of marginal and plasma cell types in PTLDs. Polymorphic PTLD cases represent EBV-induced expansion of B cells, mimicking CD40L-like activated Peri/Interfollicular CD30+ normal B-cells.

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Evolution of a True B-Cell Chronic Lymphocytic Leukaemia (B-CLL) in a Diffuse Immunocytoma after Treatment with Fludarabine.

Blood ◽

10.1182/blood.v106.11.4973.4973 ◽

2005 ◽

Vol 106 (11) ◽

pp. 4973-4973

Author(s):

Paolo Bernasconi ◽

Marco Paulli ◽

Ester Orlandi ◽

Vittorio Perfetti ◽

Ilaria Giardini ◽

...

Keyword(s):

B Cell ◽

B Lymphocytes ◽

Cell Population ◽

Plasma Cells ◽

Opportunistic Infections ◽

Normal Karyotype ◽

Prognostic Impact ◽

Clonal Cell ◽

Lp Cells

Abstract In B-CLL CD5, CD23 antigen-experienced B-cell are the neoplastic population. However, a small proportion of patients may present a monoclonal component (MC) and lymphoplasmacytoid (LP) cells, different from those CD5,CD23 negative of immunocytoma. Fludarabine, a very effective drug in the treatment of B-CLL and indolent lymphomas, may favour opportunistic infections and secondary cancers due to intense and prolonged immunosuppression. Herein we report on a 58-year old woman with night sweats and weakness. Physical examination revealed small peripheral lymphoadenopathies. WBC were 17.2x109/l; the marrow was infiltrated by 52% small mature lymphocytes presenting a monotypic kappa light chain restriction and CD5, CD19, CD20, CD23, CD38, ZAP70 antigens. Cytogenetics revealed a normal karyotype, whereas FISH detected 13q- in 48% nuclei. A diagnosis of B-CLL in Rai stage IA without any MC was made. The patient did not receive any treatment and nine months later developed multiple adenopathies and a progressive splenomegaly. WBC were 32.9x109/l; the marrow was infiltrated by 78% mature B lymphocytes and 5% LP cells sharing the original phenotype. Therefore a progression in Rai stage III had occurred and the patient started treatment with oral fludarabine (40mg/m2/day for 5 days, repeated every 4 weeks) which was well tolerated. A clinical response was achieved after fourth courses, with disappearance of adenopathies and splenomegaly, and normalization of peripheral blood. Few days later the patient complained of itching. Her blood tests revealed eosinophilia (42%), but twenty days later an IgG kappa MC (4,3g/dl) became apparent. Ten days later the patient was admitted to our ward because of liver, spleen and lymphonode enlargement and a doubled MC (10.9g/dl). The bone marrow was infiltrated by 60% mature B lymphocytes with the original phenotype and 20% plasma cells with cytoplasmic IgM and IgG, a monotypic restriction for kappa light chains, CD38, CD138, CD56 antigens. FISH showed that both cell populations presented a 13q- and the analysis of the CDRIII region detected only one clonal cell population with mutated Ig(V) genes. In addition a lymphonode biopsy showed a parafollicular and nodular pattern of infiltration by CD20+, CD79a+, CD5+, CD23 +/−, CD10- and Bcl1- B-cells, showing a trend towards differentiation in secreting elements, and by 18% prolymphocytes and paraimmunoblasts grouped in nodular aggregates. In conclusion, the lymphonode histology addressed to an immunocytoma in initial evolution in Richter’s syndrome. Our patient emphasizes the overlapping features between B-CLL and lymphoplasmacytoid lymphoma (LPL) since imunophenotypic, FISH, CDIII analysis demonstrated that the initial clonal cell population was able to undergo in vivo isotype switching to IgG, giving rise to LP elements and plasma cells producing high IgG levels. In addition, the correctness of the initial diagnosis is confirmed by the absence of t(9;14), a rearrangement typical of LPL, and the presence of 13q-, which prognostic impact in B-CLL with LP differentiation may be different than in typical B-CLL. The influence of fludarabine on the differentiation process remains unclear.

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Autoantibody Production during Chronic Graft-Versus-Host Disease Does Not Associate with Long-Term Persistence of Host B Cells in Humans

Blood ◽

10.1182/blood.v112.11.1180.1180 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1180-1180

Author(s):

Simona Piemontese ◽

Zulma Magnani ◽

Jacopo Peccatori ◽

Claudio Bordignon ◽

Chiara Bonini ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

B Lymphocytes ◽

Graft Versus Host Disease ◽

Cell Depletion ◽

B Cell Depletion ◽

Autoantibody Production ◽

Graft Versus Host

Abstract Background. Chronic graft-versus-host disease (cGvHD) is a common complication of allogeneic hemopoietic cell transplantation (allo-HCT). The pathogenesis of cGvHD is poorly understood. In cGvHD, the homeostasis of B lymphocytes is perturbed, as demonstrated by the production of autoantibodies. B-cell depletion with monoclonal antibodies (mAb) interferes with autoantibody production and ameliorates signs and symptoms of cGvHD. In mouse models, cGvHD and autoantibodies associate with the long-term persistence of host B cells after allo-HCT (Sylvain Perruche et al., Transplantation 2006). It has been postulated that host B cells may present alloantigens to donor T cells and, in turn, receive help for autoantibody production. This could be crucial to the pathogenesis of cGvHD. Aim. To investigate whether the long-term persistence of host B lymphocytes is associated with cGvHD and autoantibodies in humans. Patients and methods. We recruited 13 consecutive patients with active cGvHD (4 mild, 5 moderate, 4 severe according to NIH classification) with a median time of onset of 6 months (range 3–36) from HLA-identical sibling (9 patients) and HLA-matched unrelated (4) allo-HCT. As controls, we chose 10 patients that underwent HLAidentical sibling (2), HLA-matched unrelated (5) or haploidentical (3) allo-HCT and never experienced cGvHD. In the two groups, we studied: circulating autoantibodies, including anti-nuclear (ANA), anti-DNA, anti-extractable nuclear antigen, anti-beta2 glycoprotein, anti-neutrophil cytoplasm, anti-thyroid, anti-mytocondria antibodies, rheumatoid factor, absolute numbers of T (CD3+, CD4+, CD8+), conventional B (CD19+), B1 (CD5+/CD19+) and NK cells (CD16+/CD56+) in the graft and in the peripheral blood, microchimerism by short-tandem repeats (STR) on B, T and myeloid cells purified by immunomagnetic cell sorting (sensitivity 0,01%). Results. Patients with cGvHD had high-titer circulating ANA (>1:160) more frequently than controls (54% versus 10%, P<0,05). All other autoantibodies were negative. Peripheral T-cell counts were lower in patients with cGvHD than in controls (for CD8+ cells P<0,05). This was not due to a difference in the absolute numbers of T lymphocytes within the graft between the two groups. Peripheral counts of conventional B and B1 cells in patients with cGvHD were similar to controls. Autoantibodies and cGvHD were not associated with the persistence of host B lymphocytes, since the analysis of STR on purified B cells revealed that they were all of donor origin. T and myeloid cells were also of donor origin. Of interest, in univariate analysis, in vivo B-cell depletion with mAb for the prophylaxis against Epstein-Barr virus-related lymphoproliferative disease showed a trend towards a lower risk of cGvHD (P=0,06). Conclusions. This study indicates that autoantibody production during cGvHD does not associate with long-term persistence of host B cells in humans. Moreover, it suggests that the early depletion of donor B lymphocytes in vivo may be effective for GvHD prophylaxis

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Prolonged survival in vivo of unprimed B cells responsive to a T-independent antigen.

Journal of Experimental Medicine ◽

10.1084/jem.161.6.1581 ◽

1985 ◽

Vol 161 (6) ◽

pp. 1581-1586 ◽

Cited By ~ 12

Author(s):

Y Ron ◽

J Sprent

Keyword(s):

Lymph Node ◽

B Cells ◽

B Cell ◽

B Lymphocytes ◽

Thoracic Duct ◽

Thoracic Duct Lymph ◽

Cell Transfer ◽

Adult Mice ◽

Duct Lymph

Despite earlier evidence to the contrary, it has recently been claimed that most B lymphocytes, including lymph node (LN) and thoracic duct B cells, are short-lived cells of recent marrow origin. To seek direct information on this question, we transferred unprimed LN or thoracic duct B cells from normal mice to xid mice, i.e., mice unresponsive to the T-independent antigen, trinitrophenyl (TNP)-Ficoll. At varying periods after B cell transfer the recipients were challenged with TNP-Ficoll; anti-TNP plaque-forming cells were assayed in the spleen 6 d later. The results showed that the B cell recipients retained responsiveness to TNP-Ficoll for at least 3 mo after transfer. Responsiveness increased within the first 3 wk but then remained relatively constant. These findings imply that, at least for TNP-Ficoll-reactive cells, B cells residing in LN and thoracic duct lymph are not short-lived cells of recent marrow. Indeed, the data suggest that once the pool of recirculating B cells is fully formed in adult mice, further input of newly formed cells from the marrow into the recirculating pool is very limited.

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BCR-mediated uptake of antigen linked to TLR9 ligand stimulates B-cell proliferation and antigen-specific plasma cell formation

Blood ◽

10.1182/blood-2008-10-185421 ◽

2009 ◽

Vol 113 (17) ◽

pp. 3969-3977 ◽

Cited By ~ 102

Author(s):

Julia Eckl-Dorna ◽

Facundo D. Batista

Keyword(s):

Cell Proliferation ◽

B Cells ◽

B Cell ◽

Plasma Cells ◽

Cell Formation ◽

Specific Class ◽

B Cell Differentiation ◽

Enhanced Production ◽

Stimulation Of

Abstract The activation of Toll-like receptor 9 (TLR9) expressed within B cells is associated with enhanced humoral immunity. However the role of TLR9 in the stimulation of B-cell responses, and more specifically in shaping the outcome of B-cell differentiation, remains unclear. Here, we observed that immunization with the TLR9 agonist CpG linked to protein antigen gave rise to enhanced production of antigen-specific class-switched antibodies in vivo. Unlike dendritic cells, B cells are unable to acquire these conjugates by macropinocytosis and instead depend on uptake through a signaling-competent B-cell receptor (BCR), provided the overall BCR-antigen avidity exceeds a defined threshold. The resultant stimulation of intrinsic TLR9 leads to enhanced antigen-specific B-cell proliferation and differentiation to form extrafollicular plasma cells. Thus, the direct conjugation of antigen and CpG reveals a mechanism that may operate during the initiation of primary immune responses, and may prove useful as a strategy for the design of adjuvants suitable for vaccinations.

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IgM and IgD B cell receptors differentially respond to endogenous antigens and control B cell fate

eLife ◽

10.7554/elife.35074 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 23

Author(s):

Mark Noviski ◽

James L Mueller ◽

Anne Satterthwaite ◽

Lee Ann Garrett-Sinha ◽

Frank Brombacher ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Fate ◽

Plasma Cells ◽

Autoreactive B Cells ◽

Binding Domains ◽

Bcr Signaling ◽

And Control

Naive B cells co-express two BCR isotypes, IgM and IgD, with identical antigen-binding domains but distinct constant regions. IgM but not IgD is downregulated on autoreactive B cells. Because these isotypes are presumed to be redundant, it is unknown how this could impose tolerance. We introduced the Nur77-eGFP reporter of BCR signaling into mice that express each BCR isotype alone. Despite signaling strongly in vitro, IgD is less sensitive than IgM to endogenous antigen in vivo and developmental fate decisions are skewed accordingly. IgD-only Lyn−/− B cells cannot generate autoantibodies and short-lived plasma cells (SLPCs) in vivo, a fate thought to be driven by intense BCR signaling induced by endogenous antigens. Similarly, IgD-only B cells generate normal germinal center, but impaired IgG1+ SLPC responses to T-dependent immunization. We propose a role for IgD in maintaining the quiescence of autoreactive B cells and restricting their differentiation into autoantibody secreting cells.

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Occurrence of cytochrome b558 in B-cell lineage of human lymphocytes

Blood ◽

10.1182/blood.v75.2.458.bloodjournal752458 ◽

1990 ◽

Vol 75 (2) ◽

pp. 458-461

Author(s):

S Kobayashi ◽

S Imajoh-Ohmi ◽

M Nakamura ◽

S Kanegasaki

Keyword(s):

B Cells ◽

Cell Surface ◽

B Cell ◽

B Lymphocytes ◽

Plasma Cells ◽

Human Lymphocytes ◽

Cell Lineage ◽

Superoxide Generation ◽

Cell Stage ◽

Cytochrome B558

Cytochrome b558, involved in superoxide generation in phagocytes, was found to be expressed on the cell surface of most normal peripheral B lymphocytes. The cytochrome was not found on the surface of peripheral T lymphocytes, natural killer cells, or peripheral lymphocytes derived from patients with X-linked chronic granulomatous disease. On stimulation, at least half of peripheral B lymphocytes could generate superoxide anion as detected by superoxide dismutase-sensitive nitroblue tetrazorium reduction. Cytochrome b558 was not present on the surface of pre-pre B cells or pre-B cells, but did appear at the early B-cell stage. It disappeared from the B-cell surface during terminal differentiation to plasma cells. The transient expression of the cytochrome in B-cell lineage may indicate that superoxide generation is important for the function of these cells at certain stages.

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IRF4 controls the positioning of mature B cells in the lymphoid microenvironments by regulating NOTCH2 expression and activity

Journal of Experimental Medicine ◽

10.1084/jem.20131026 ◽

2013 ◽

Vol 210 (13) ◽

pp. 2887-2902 ◽

Cited By ~ 36

Author(s):

Giorgia Simonetti ◽

Amanda Carette ◽

Kathryn Silva ◽

Haowei Wang ◽

Nilushi S. De Silva ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Plasma Cells ◽

Developmental Stages ◽

Nuclear Translocation ◽

Inhibitory Antibody ◽

Class Switch ◽

Elevated Protein ◽

Interferon Regulatory Factor 4

The transcription factor interferon regulatory factor-4 (IRF4) is expressed in B cells at most developmental stages. In antigen-activated B cells, IRF4 controls germinal center formation, class-switch recombination, and the generation of plasma cells. Here we describe a novel function for IRF4 in the homeostasis of mature B cells. Inducible deletion of irf4 specifically in B cells in vivo led to the aberrant accumulation of irf4-deleted follicular B cells in the marginal zone (MZ) area. IRF4-deficient B cells showed elevated protein expression and activation of NOTCH2, a transmembrane receptor and transcriptional regulator known to be required for MZ B cell development. Administration of a NOTCH2-inhibitory antibody abolished nuclear translocation of NOTCH2 in B cells within 12 h and caused a rapid and progressive disintegration of the MZ that was virtually complete 48 h after injection. The disappearance of the MZ was accompanied by a transient increase of MZ-like B cells in the blood rather than increased B cell apoptosis, demonstrating that continued NOTCH2 activation is critical for the retention of B cells in the MZ. Our results suggest that IRF4 controls the positioning of mature B cells in the lymphoid microenvironments by regulating NOTCH2 expression. These findings may have implications for the understanding of B cell malignancies with dysregulated IRF4 and NOTCH2 activity.

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