scholarly journals Prolactin Rescues Immature B Cells from Apoptosis-Induced BCR-Aggregation through STAT3, Bcl2a1a, Bcl2l2, and Birc5 in Lupus-Prone MRL/lpr Mice

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 316 ◽  
Author(s):  
Rocio Flores-Fernández ◽  
Angélica Aponte-López ◽  
Mayra C. Suárez-Arriaga ◽  
Patricia Gorocica-Rosete ◽  
Alberto Pizaña-Venegas ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Chromatin Immunoprecipitation ◽  
Prolactin Receptor ◽  
Apoptosis Resistance ◽  
Clonal Deletion ◽  
Prolactin Signaling ◽  
Stat3 Phosphorylation ◽  
Immature B Cells

Self-reactive immature B cells are eliminated through apoptosis by tolerance mechanisms, failing to eliminate these cells results in autoimmune diseases. Prolactin is known to rescue immature B cells from B cell receptor engagement-induced apoptosis in lupus-prone mice. The objective of this study was to characterize in vitro prolactin signaling in immature B cells, using sorting, PCR array, RT-PCR, flow cytometry, and chromatin immunoprecipitation. We found that all B cell maturation stages in bone marrow express the prolactin receptor long isoform, in both wild-type and MRL/lpr mice, but its expression increased only in the immature B cells of the latter, particularly at the onset of lupus. In these cells, activation of the prolactin receptor promoted STAT3 phosphorylation and upregulation of the antiapoptotic Bcl2a1a, Bcl2l2, and Birc5 genes. STAT3 binding to the promoter region of these genes was confirmed through chromatin immunoprecipitation. Furthermore, inhibitors of prolactin signaling and STAT3 activation abolished the prolactin rescue of self-engaged MRL/lpr immature B cells. These results support a mechanism in which prolactin participates in the emergence of lupus through the rescue of self-reactive immature B cell clones from central tolerance clonal deletion through the activation of STAT3 and transcriptional regulation of a complex network of genes related to apoptosis resistance.

Defective B-cell-negative selection and terminal differentiation in the ICF syndrome

Blood ◽  
2004 ◽  
Vol 103 (7) ◽  
pp. 2683-2690 ◽  
Author(s):  
Carla E. Blanco-Betancourt ◽  
Anne Moncla ◽  
Michèle Milili ◽  
Yun Liang Jiang ◽  
Evani M. Viegas-Péquignot ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Negative Selection ◽  
Dna Methyltransferase ◽  
Plasma Cells ◽  
Immunoglobulin E ◽  
Variable Regions ◽  
Icf Syndrome ◽  
Immature B Cells

Abstract Immunodeficiency, centromeric region instability, and facial anomalies (ICF) syndrome is a rare autosomal recessive disease. Mutations in the DNA methyltransferase 3B (DNMT3B) gene are responsible for most ICF cases reported. We investigated the B-cell defects associated with agammaglobulinemia in this syndrome by analyzing primary B cells from 4 ICF patients. ICF peripheral blood (PB) contains only naive B cells; memory and gut plasma cells are absent. Naive ICF B cells bear potentially autoreactive long heavy chain variable regions complementarity determining region 3's (VHCDR3's) enriched with positively charged residues, in contrast to normal PB transitional and mature B cells, indicating that negative selection is impaired in patients. Like anergic B cells in transgenic models, newly generated and immature B cells accumulate in PB. Moreover, these cells secrete immunoglobulins and exhibit increased apoptosis following in vitro activation. However, they are able to up-regulate CD86, indicating that mechanisms other than anergy participate in silencing of ICF B cells. One patient without DNMT3B mutations shows differences in immunoglobulin E (IgE) switch induction, suggesting that immunodeficiency could vary with the genetic origin of the syndrome. In this study, we determined that negative selection breakdown and peripheral B-cell maturation blockage contribute to agammaglobulinemia in the ICF syndrome. (Blood. 2004;103:2683-2690)

scholarly journals In vitro tolerance induction of neonatal murine B cells.

1976 ◽  
Vol 143 (6) ◽  
pp. 1327-1340 ◽  
Author(s):  
E S Metcalf ◽  
N R Klinman
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Specific Interaction ◽  
Tolerance Induction ◽  
Antigen Receptors ◽  
Spleen Cells ◽  
Protein Conjugates ◽  
Immature B Cells

The susceptibility of neonatal and adult B lymphocytes to tolerance induction was analyzed by a modification of the in vitro splenic focus technique. This technique permits stimulation of individual hapten-specific clonal precursor cells from both neonatal and adult donors. Neonatal or adult BALB/c spleen cells were adoptively transferred into irradiated, syngeneic, adult recipients which had been carrier-primed to hemocyanin (Hy), thus maximizing stimulation to the hapten 2,4-dinitrophenyl coupled by Hy (DNP-Hy). Cultures were initially treated with DNP on several heterologous (non-Hy) carriers and subsequently stimulated with DNP-Hy. Whereas the responsiveness of adult B cells was not diminished by pretreatment with any DNP conjugate, the majority of the neonatal B-cell response was abolished by in vitro culture with all of the DNP-protein conjugates. During the 1st wk of life, the ability to tolerize neonatal splenic B cells progressively decreased. Thus, tolerance in this system is: (a) restricted to B cells early in development; (b) established by both tolerogens and immunogens; (c) achieved at low (10(-9) M determinant) antigen concentrations; and (d) highly specific, discriminating between DNP- and TNP-specific B cells. We conclude that: (a) B lymphocytes, during their development, mature through a stage in which they are extremely susceptible to tolerogenesis; (b) the specific interaction of B-cell antigen receptors with multivalent antigens, while irrelevant to mature B cells, is tolerogenic to neonatal (immature) B cells unless antigen is concomitantly recognized by primed T cells; and (c) differences in the susceptibility of immature and mature B lymphocytes to tolerance induction suggest intrinsic differences between neonatal and adult B cells and may provide a physiologically relevant model for the study of tolerance to self-antigens.

scholarly journals Prolactin Rescues Immature B-Cells from Apoptosis Induced by B-Cell Receptor Cross-Linking

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Rocio Flores-Fernández ◽  
Francisco Blanco-Favela ◽  
Ezequiel M. Fuentes-Pananá ◽  
Luis Chávez-Sánchez ◽  
Patricia Gorocica-Rosete ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Lupus Erythematosus ◽  
Receptor Interaction ◽  
Cross Linking ◽  
Relative Expression ◽  
Wehi 231 ◽  
Immature B Cells ◽  
The Cross

Prolactin has an immunomodulatory effect and has been associated with B-cell-triggered autoimmune diseases, such as systemic lupus erythematosus (SLE). In mice that develop SLE, the PRL receptor is expressed in early bone marrow B-cells, and increased levels of PRL hasten disease manifestations, which are correlated with a reduction in the absolute number of immature B-cells. The aim of this work was to determine the effect of PRL in anin vitrosystem of B-cell tolerance using WEHI-231 cells and immature B-cells from lupus prone MRL/lpr mice. WEHI-231 cells express the long isoform of the PRL receptor, and PRL rescued the cells from cell death by decreasing the apoptosis induced by the cross-linking of the B-cell antigen receptor (BCR) as measured by Annexin V and active caspase-3. This decrease in apoptosis may have been due to the PRL and receptor interaction, which increased the relative expression of antiapoptotic Bcl-xL and decreased the relative expression of proapoptotic Bad. In immature B-cells from MRL/lpr mice, PRL increased the viability and decreased the apoptosis induced by the cross-linking of BCR, which may favor the maturation of self-reactive B-cells and contribute to the onset of disease.

scholarly journals Role of IgD in the immune response and tolerance. I. Anti-delta pretreatment facilitates tolerance induction in adult B cells in vitro.

1977 ◽  
Vol 146 (6) ◽  
pp. 1473-1483 ◽  
Author(s):  
D W Scott ◽  
J E Layton ◽  
G J Nossal
Keyword(s):  
Immune Response ◽  
B Cells ◽  
B Cell ◽  
Tolerance Induction ◽  
Spleen Cells ◽  
Cell Compartment ◽  
Immature B Cells

Adult spleen cells from C57BL.Ige mice, which generally are resistant to in vitro tolerance induction in the B-cell compartment, became hyporesponsive (tolerant) when cultured with antigen in the presence of an anti-allotype serum. Both antigen and anti-delta had to be present for this effect, which was hapten-specific and did not occur in C57BL/L mice, which lack the Ig5-1 allotype of the delta-chain detected in this system. Preculture with anti-mu serum plus antigen, in contrast, did not cause tolerance induction in adult spleen B cells of either strain. These results suggest that the surface IgD may act as a failsafe receptor to prevent tolerance induction in adult B cells. Tolerance studies with spleen cells from mice with markedly reduced numbers of IgD+ve cells, because of regimen of repeated injections of anti-delta serum beginning at birth (delta-suppressed mice), confirmed the importance of membrane IgD in preventing tolerance, because such delta-suppressed mice were hypersusceptible to tolerance by antigen alone. Inasmuch as immature B cells lack IgD on their surface, these studies suggest that acquisition of IgD is an important maturational step in the ability of murine B cells to discriminate tolerogenic and immunogenic signals.

scholarly journals Addition of constitutive c-myc expression to Abelson murine leukemia virus changes the phenotype of the cells transformed by the virus from pre-B-cell lymphomas to plasmacytomas.

1993 ◽  
Vol 13 (4) ◽  
pp. 2578-2585 ◽  
Author(s):  
E M Weissinger ◽  
H Mischak ◽  
J Goodnight ◽  
W F Davidson ◽  
J F Mushinski
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Leukemia Virus ◽  
B Cell Lymphomas ◽  
Immature B Cells ◽  
Abelson Murine Leukemia Virus ◽  
Murine Leukemia

Abelson murine leukemia virus (A-MuLV), a retrovirus that expresses the v-abl oncogene, characteristically induces pre-B-cell lymphomas following in vivo infection of BALB/c mice or in vitro infection of suspensions of fetal liver or bone marrow cells. ABL-MYC, a retrovirus that expresses both v-abl and c-myc, induces solely plasmacytomas in BALB/c mice. To investigate how the addition of overexpression of c-myc to that of v-abl accomplishes this dramatic change in the phenotype of the cells transformed by these closely related retroviruses, we utilized helper-free A-MuLV (psi 2) and ABL-MYC (psi 2) in vitro to infect suspensions of cells from different lymphoid tissues and purified immature and purified mature B cells. As expected, A-MuLV(psi 2) induced only pre-B-cell lymphomas in vivo and in vitro when immature B cells were present. ABL-MYC(psi 2), on the other hand, produced only plasmacytomas, even when purified immature B lymphocytes were infected in vitro. Although the A-MuLV(psi 2)-induced pre-B-cell lymphomas express easily detectable levels of c-myc mRNA, maturation into more-mature forms of B lymphocytes is blocked. The constitutively overexpressed c-myc in the ABL-MYC retrovirus abrogates this block, permits maturation of infected immature B cells, and yields transformed plasma cells.

CD19 regulates positive selection and maturation in B lymphopoiesis: lack of CD19 imposes developmental arrest of immature B cells and consequential stimulation of receptor editing

Blood ◽  
2005 ◽  
Vol 105 (8) ◽  
pp. 3247-3254 ◽  
Author(s):  
Eran Diamant ◽  
Zohar Keren ◽  
Doron Melamed
Keyword(s):  
B Cells ◽  
Positive Selection ◽  
B Cell ◽  
Receptor Editing ◽  
B Lymphopoiesis ◽  
Developmental Arrest ◽  
Surface Marker Expression ◽  
Immature B Cells

AbstractLigand-independent signals that are produced by the B-cell antigen receptor (BCR) confer an important positive selection checkpoint for immature B cells. Generation of inappropriate signals imposes developmental arrest of immature B cells, though the fate of these cells has not been investigated. Studies have shown that the lack of CD19 results in inappropriate signaling. In immunoglobulin transgenic mice, this inappropriate signaling impairs positive selection and stimulates receptor editing. Here, we studied the extent and significance of receptor editing in CD19-regulated positive selection of normal, nontransgenic B lymphopoiesis, using our bone marrow culture system. We found that the lack of CD19 resulted in elevated tonic signaling and impaired maturation, as revealed by surface marker expression and by functional assays. Immature CD19-/- B cells did not suppress RAG and underwent intensive receptor editing attempts in culture. Finally, in vivo analysis of light-chain isotype expression and Jκ use in CD19-/- mice validated our in vitro observations. Our results suggest that CD19 has an important function in regulating positive selection and maturation of nontransgenic B-cell precursors and that receptor editing is an important salvage mechanism for immature B cells that fail positive selection. (Blood. 2005;105:3247-3254)

scholarly journals Prolonged survival of B-lineage acute lymphoblastic leukemia cells is accompanied by overexpression of bcl-2 protein

Blood ◽  
1993 ◽  
Vol 81 (4) ◽  
pp. 1025-1031 ◽  
Author(s):  
D Campana ◽  
E Coustan-Smith ◽  
A Manabe ◽  
M Buschle ◽  
SC Raimondi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
B Cells ◽  
B Cell ◽  
Lymphoblastic Leukemia ◽  
Prolonged Survival ◽  
Allogeneic Bone ◽  
Immature B Cells ◽  
B Lineage

Overexpression of bcl-2 delays the onset of apoptosis in lymphohematopoietic cells. We measured levels of bcl-2 protein in normal and leukemic human B-cell progenitors with a specific monoclonal antibody and flow cytometry. Normal immature B cells had low levels of bcl-2 protein; the intensity of fluorescence, expressed as molecules of soluble fluorochrome per cell, within CD10+ cells was 3,460 +/- 1,050 (mean +/- SD; 5 samples). In 16 cases of B-lineage acute lymphoblastic leukemia (ALL), cells had levels of bcl-2 that were strikingly higher than those of their normal counterparts (33,560 +/- 14,570; P < .001 by t-test analysis). We next investigated whether the intensity of bcl-2 expression correlated with the resistance of immature B cells to in vitro culture. In 12 cases of B-lineage ALL, the cells recovered after 7 days of culture on allogeneic bone marrow stromal layers were 69% to 178% (median, 95.5%) of those originally seeded. Prolonged survival of leukemic cells in vitro was observed even in the absence of stromal layers in 6 of these 12 cases; the intensity of bcl-2 protein expression in these cases was 45,000 +/- 13,270, compared with 21,500 +/- 7,260 in the 6 cases in which greater than 99.5% of cells rapidly died by apoptosis under the same culture conditions (P = .003). Five immature B-cell lines, continuously growing in the absence of stroma, had the highest bcl-2 expression (79,400 +/- 20,330). By contrast, most normal CD19+, sIg-immature B cells died despite the presence of bone marrow stromal layers; 9.7% to 28.2% were recovered after 7 days of culture in three experiments. We conclude that abnormal bcl-2 gene expression influences the survival ability of B-cell progenitors. This may contribute to leukemogenesis and explain the aptitude of leukemic lymphoblasts to expand outside the bone marrow microenvironment.

Addition of constitutive c-myc expression to Abelson murine leukemia virus changes the phenotype of the cells transformed by the virus from pre-B-cell lymphomas to plasmacytomas

1993 ◽  
Vol 13 (4) ◽  
pp. 2578-2585
Author(s):  
E M Weissinger ◽  
H Mischak ◽  
J Goodnight ◽  
W F Davidson ◽  
J F Mushinski
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Leukemia Virus ◽  
B Cell Lymphomas ◽  
Immature B Cells ◽  
Abelson Murine Leukemia Virus ◽  
Murine Leukemia

Abelson murine leukemia virus (A-MuLV), a retrovirus that expresses the v-abl oncogene, characteristically induces pre-B-cell lymphomas following in vivo infection of BALB/c mice or in vitro infection of suspensions of fetal liver or bone marrow cells. ABL-MYC, a retrovirus that expresses both v-abl and c-myc, induces solely plasmacytomas in BALB/c mice. To investigate how the addition of overexpression of c-myc to that of v-abl accomplishes this dramatic change in the phenotype of the cells transformed by these closely related retroviruses, we utilized helper-free A-MuLV (psi 2) and ABL-MYC (psi 2) in vitro to infect suspensions of cells from different lymphoid tissues and purified immature and purified mature B cells. As expected, A-MuLV(psi 2) induced only pre-B-cell lymphomas in vivo and in vitro when immature B cells were present. ABL-MYC(psi 2), on the other hand, produced only plasmacytomas, even when purified immature B lymphocytes were infected in vitro. Although the A-MuLV(psi 2)-induced pre-B-cell lymphomas express easily detectable levels of c-myc mRNA, maturation into more-mature forms of B lymphocytes is blocked. The constitutively overexpressed c-myc in the ABL-MYC retrovirus abrogates this block, permits maturation of infected immature B cells, and yields transformed plasma cells.

CLL B Cell Interaction with Bone Biopsy Generated Marrow Stromal Elements Enhances Their Apoptosis Resistance in Association with an Angiogenic Switch.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1914-1914
Author(s):  
Neil E. Kay ◽  
Ann K. Strege ◽  
Tait D. Shanafelt ◽  
Nancy D. Bone ◽  
Yean K. Lee
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Growth Factor ◽  
B Cell ◽  
Culture Medium ◽  
Bone Biopsy ◽  
Cell Interaction ◽  
Apoptosis Resistance ◽  
Angiogenic Switch

Abstract CLL B cells have significant opportunity to interact with multiple tissue sites that are critical to the leukemic cell’s ability to survive. The marrow stromal elements (MSE) in B-chronic lymphocytic leukemia (B-CLL) are very likely one of the most frequently involved in this important cell-cell interaction. We have utilized a novel bone marrow biopsy technique (Alvi S, Leuk Res, 2001) that consistently generates robust and long-lived marrow stromal elements from B-CLL patients in order to study CLL B cell apoptosis status in relation to angiogenic factors. The latter angiogenic parameters were studied as we have previously detected both a VEGF based autocrine pathway for CLL B cells and found an extensive neovascularization in CLL marrows. Bone biopsy material from B-CLL patients (N=25) yielded active cell growth in vitro which expresses the features of marrow stromal constituents. Importantly, these marrow constituents were able to be sustained for several months (range, 3 – 52 weeks) and also could be transferred repetitively to new culture dishes. CLL B cells were able to spontaneously release VEGF and TSP-1 as we have previously published (Kay N, Leukemia, 2002). MSE from CLL marrow (N=5) were found to spontaneously secrete basic fibroblast growth factor (bFGF) 5.2 ± 0.9 pg/ml (mean ± one SEM), vascular endothelial growth factor (VEGF) 162 ± 48 pg/ml and thrombospondin-1 (TSP-1) 90 ± 26 ng/ml into the culture medium. Significant reduction in CLL B cell spontaneous apoptosis,measured by Annexin/PI incorporation (mean ± one SEM), was seen after exposure to the CLL MSE (N=6) both after 24 hours (i.e., Non exposed CLL B cells vs. MSE co-cultured CLL B cells was 70% ± 9 vs. 39% ± 10 respectively for 24 hours, p < 0.01) and 72 hrs (data not shown). Exposure to MSE also enhanced the in vitro drug resistance of CLL B cells to chlorambucil (C). Thus, for CLL B cells (n=5) cultured with CLL MSE and C (1μM) for 24 hours, apoptosis levels were reduced from 58.8% ± 7.8 for B cells without MSE compared to 39.8% ± 3.8 for B cells exposed to MSE. Apoptosis resistance enhancement for CLL B cells was found to be similar with normal marrow stromal elements and a stromal cell line (HS-5). Anti-apoptotic proteins detected by immunoblot were found to increase for CLL B cells exposed to stromal elements from bone marrow biopsies and included Mcl-1, Bcl-2, Survivin and XIAP. While Mcl-1 was found to increase, there was a decrease in Bcl-2 and Survivin detected by immunoblot when CLL B cells were exposed to the HS-5 cell line implying a different mechanism for apoptosis enhancement. When CLL B cells were added to CLL MSE for 24 hours, there was an increase in VEGF released into the culture medium (283 pg ± 114 for the co-culture vs. 14 pg ± 1.7 and 162 pg ± 48.8 for CLL B cells and MSE cultured alone respectively). In contrast co-culture of CLL B cells with MSE (24 hours) resulted in less TSP-1 in the culture medium than expected (106 ng ± 28.8 for co-cultured CLL and MSE cells vs. 89.5 ng ± 27.5 and 90.9 ng ± 26.4 for CLL B cells and MSE cultured alone respectively). This change in the ratio of pro- to anti-angiogenic factors favoring VEGF strongly suggests that CLL B cell interaction with stroma can facilitate an angiogenic switch and may be linked to disease progression. In addition, we have found that bone biopsies from B-CLL patients can generate long-term marrow stromal elements that are able to reliably enhance CLL B cell apoptosis resistance.

scholarly journals Ras activation of Erk restores impaired tonic BCR signaling and rescues immature B cell differentiation

2010 ◽  
Vol 207 (3) ◽  
pp. 607-621 ◽  
Author(s):  
Sarah L. Rowland ◽  
Corinne L. DePersis ◽  
Raul M. Torres ◽  
Roberta Pelanda
Keyword(s):  
B Cells ◽  
B Cell ◽  
Cell Survival ◽  
Constitutive Expression ◽  
Cell Development ◽  
B Cell Development ◽  
B Cell Differentiation ◽  
Bcr Signaling ◽  
Immature B Cells

B cell receptors (BCRs) generate tonic signals critical for B cell survival and early B cell development. To determine whether these signals also mediate the development of transitional and mature B cells, we examined B cell development using a mouse strain in which nonautoreactive immunoglobulin heavy and light chain–targeted B cells express low surface BCR levels. We found that reduced BCR expression translated into diminished tonic BCR signals that strongly impaired the development of transitional and mature B cells. Constitutive expression of Bcl-2 did not rescue the differentiation of BCR-low B cells, suggesting that this defect was not related to decreased cell survival. In contrast, activation of the Ras pathway rescued the differentiation of BCR-low immature B cells both in vitro and in vivo, whereas extracellular signal-regulated kinase (Erk) inhibition impaired the differentiation of normal immature B cells. These results strongly suggest that tonic BCR signaling mediates the differentiation of immature into transitional and mature B cells via activation of Erk, likely through a pathway requiring Ras.

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