18 research outputs found

    Image_3_Plasma Cell Alloantigen 1 and IL-10 Secretion Define Two Distinct Peritoneal B1a B Cell Subsets With Opposite Functions, PC1high Cells Being Protective and PC1low Cells Harmful for the Growing Fetus.jpg

    No full text
    <p>B cells possess various immuno regulatory functions. However, research about their participation in tolerance induction toward the fetus is just emerging. Accumulating evidence supports the idea that B cells can play seemingly conflicting roles during pregnancy, either protecting or harming the fetus. Previous findings indicated the presence of two different peritoneal B cell subsets, defined by the expression of the plasma cell alloantigen 1 (PC1) and with distinct immune modulatory functions. Here, we aimed to study the participation of these two B cell subsets, on pregnancy outcome in a murine model of disturbed fetal tolerance. The frequencies and cell numbers of peritoneal and splenic CD19<sup>+</sup>IL-10<sup>+</sup> and CD19<sup>+</sup>CD5<sup>+</sup>IL-10<sup>+</sup>PC1<sup>+</sup> cells were assessed in virgin as well as normal pregnant (NP) and abortion-prone (AP) females during the course of gestation. Peritoneal PC1<sup>low</sup> or PC1<sup>high</sup> B1a B cells were sorted, analyzed for their ability to secrete IL-10 and adoptively transferred into NP or AP females. On gestation day (gd) 12, the abortion rate as well as the frequencies and cell numbers of regulatory T cells, TH1 and TH17 cells were determined in spleens and decidua. In addition, mRNA expression of IL-10, TGF-β, IFN-γ, and TNF-α was analyzed in decidual tissue. Peritoneal CD19<sup>+</sup>IL-10<sup>+</sup> and CD19<sup>+</sup>CD5<sup>+</sup>IL-10<sup>+</sup>PC1<sup>+</sup> frequencies fluctuated during the progression of normal pregnancies while no significant changes were observed in spleen. AP females showed significantly reduced frequencies of both B cell populations and exhibited an altered peritoneal PC1<sup>high</sup>/PC1<sup>low</sup> ratio at gd10. Adoptive transfers of PC1<sup>low</sup> B1a B cells into NP females increased the abortion rate in association with a reduced splenic regulatory T/TH17 ratio. By contrast, the transfer of PC1<sup>high</sup> B1a B cells into AP females significantly diminished the fetal rejection rate and significantly reduced the numbers of splenic TH17 cells. Our results suggest that the peritoneum harbors two distinct B1a B cell subsets that can be distinguished by their PC1 expression. Whereas PC1<sup>high</sup> B1a B cells seem to support fetal survival, PC1<sup>low</sup> cells B1a B cells may compromise fetal well-being.</p

    Image_1_Plasma Cell Alloantigen 1 and IL-10 Secretion Define Two Distinct Peritoneal B1a B Cell Subsets With Opposite Functions, PC1high Cells Being Protective and PC1low Cells Harmful for the Growing Fetus.jpg

    No full text
    <p>B cells possess various immuno regulatory functions. However, research about their participation in tolerance induction toward the fetus is just emerging. Accumulating evidence supports the idea that B cells can play seemingly conflicting roles during pregnancy, either protecting or harming the fetus. Previous findings indicated the presence of two different peritoneal B cell subsets, defined by the expression of the plasma cell alloantigen 1 (PC1) and with distinct immune modulatory functions. Here, we aimed to study the participation of these two B cell subsets, on pregnancy outcome in a murine model of disturbed fetal tolerance. The frequencies and cell numbers of peritoneal and splenic CD19<sup>+</sup>IL-10<sup>+</sup> and CD19<sup>+</sup>CD5<sup>+</sup>IL-10<sup>+</sup>PC1<sup>+</sup> cells were assessed in virgin as well as normal pregnant (NP) and abortion-prone (AP) females during the course of gestation. Peritoneal PC1<sup>low</sup> or PC1<sup>high</sup> B1a B cells were sorted, analyzed for their ability to secrete IL-10 and adoptively transferred into NP or AP females. On gestation day (gd) 12, the abortion rate as well as the frequencies and cell numbers of regulatory T cells, TH1 and TH17 cells were determined in spleens and decidua. In addition, mRNA expression of IL-10, TGF-β, IFN-γ, and TNF-α was analyzed in decidual tissue. Peritoneal CD19<sup>+</sup>IL-10<sup>+</sup> and CD19<sup>+</sup>CD5<sup>+</sup>IL-10<sup>+</sup>PC1<sup>+</sup> frequencies fluctuated during the progression of normal pregnancies while no significant changes were observed in spleen. AP females showed significantly reduced frequencies of both B cell populations and exhibited an altered peritoneal PC1<sup>high</sup>/PC1<sup>low</sup> ratio at gd10. Adoptive transfers of PC1<sup>low</sup> B1a B cells into NP females increased the abortion rate in association with a reduced splenic regulatory T/TH17 ratio. By contrast, the transfer of PC1<sup>high</sup> B1a B cells into AP females significantly diminished the fetal rejection rate and significantly reduced the numbers of splenic TH17 cells. Our results suggest that the peritoneum harbors two distinct B1a B cell subsets that can be distinguished by their PC1 expression. Whereas PC1<sup>high</sup> B1a B cells seem to support fetal survival, PC1<sup>low</sup> cells B1a B cells may compromise fetal well-being.</p

    Image_2_Plasma Cell Alloantigen 1 and IL-10 Secretion Define Two Distinct Peritoneal B1a B Cell Subsets With Opposite Functions, PC1high Cells Being Protective and PC1low Cells Harmful for the Growing Fetus.jpg

    No full text
    <p>B cells possess various immuno regulatory functions. However, research about their participation in tolerance induction toward the fetus is just emerging. Accumulating evidence supports the idea that B cells can play seemingly conflicting roles during pregnancy, either protecting or harming the fetus. Previous findings indicated the presence of two different peritoneal B cell subsets, defined by the expression of the plasma cell alloantigen 1 (PC1) and with distinct immune modulatory functions. Here, we aimed to study the participation of these two B cell subsets, on pregnancy outcome in a murine model of disturbed fetal tolerance. The frequencies and cell numbers of peritoneal and splenic CD19<sup>+</sup>IL-10<sup>+</sup> and CD19<sup>+</sup>CD5<sup>+</sup>IL-10<sup>+</sup>PC1<sup>+</sup> cells were assessed in virgin as well as normal pregnant (NP) and abortion-prone (AP) females during the course of gestation. Peritoneal PC1<sup>low</sup> or PC1<sup>high</sup> B1a B cells were sorted, analyzed for their ability to secrete IL-10 and adoptively transferred into NP or AP females. On gestation day (gd) 12, the abortion rate as well as the frequencies and cell numbers of regulatory T cells, TH1 and TH17 cells were determined in spleens and decidua. In addition, mRNA expression of IL-10, TGF-β, IFN-γ, and TNF-α was analyzed in decidual tissue. Peritoneal CD19<sup>+</sup>IL-10<sup>+</sup> and CD19<sup>+</sup>CD5<sup>+</sup>IL-10<sup>+</sup>PC1<sup>+</sup> frequencies fluctuated during the progression of normal pregnancies while no significant changes were observed in spleen. AP females showed significantly reduced frequencies of both B cell populations and exhibited an altered peritoneal PC1<sup>high</sup>/PC1<sup>low</sup> ratio at gd10. Adoptive transfers of PC1<sup>low</sup> B1a B cells into NP females increased the abortion rate in association with a reduced splenic regulatory T/TH17 ratio. By contrast, the transfer of PC1<sup>high</sup> B1a B cells into AP females significantly diminished the fetal rejection rate and significantly reduced the numbers of splenic TH17 cells. Our results suggest that the peritoneum harbors two distinct B1a B cell subsets that can be distinguished by their PC1 expression. Whereas PC1<sup>high</sup> B1a B cells seem to support fetal survival, PC1<sup>low</sup> cells B1a B cells may compromise fetal well-being.</p

    DataSheet_1_Human chorionic gonadotropin promotes murine Treg cells and restricts pregnancy-harmful proinflammatory Th17 responses.docx

    No full text
    An equilibrium between proinflammatory and anti-inflammatory immune responses is essential for maternal tolerance of the fetus throughout gestation. To study the participation of fetal tissue-derived factors in this delicate immune balance, we analyzed the effects of human chorionic gonadotropin (hCG) on murine Treg cells and Th17 cells in vitro, and on pregnancy outcomes, fetal and placental growth, blood flow velocities and remodeling of the uterine vascular bed in vivo. Compared with untreated CD4+CD25+ T cells, hCG increased the frequency of Treg cells upon activation of the LH/CG receptor. hCG, with the involvement of IL-2, also interfered with induced differentiation of CD4+ T cells into proinflammatory Th17 cells. In already differentiated Th17 cells, hCG induced an anti-inflammatory profile. Transfer of proinflammatory Th17 cells into healthy pregnant mice promoted fetal rejection, impaired fetal growth and resulted in insufficient remodeling of uterine spiral arteries, and abnormal flow velocities. Our works show that proinflammatory Th17 cells have a negative influence on pregnancy that can be partly avoided by in vitro re-programming of proinflammatory Th17 cells with hCG.</p

    Blockage of Heme Oxygenase-1 Abrogates the Protective Effect of Regulatory T Cells on Murine Pregnancy and Promotes the Maturation of Dendritic Cells

    Get PDF
    <div><p>Regulatory T cells (Treg) play an important role in fetal protection. They expand during normal pregnancy and protect fetal antigens from maternal effector cells. Their effect is associated with the up-regulation of tolerance-associated molecules at the fetal-maternal interface. Among these, Heme Oxygenase-1 (HO-1, coded by <em>Hmox1</em>) is of special importance as its blockage correlates with increased abortion rates and its up-regulation positively affects pregnancy outcome. Here, we aimed to investigate whether the protective effect of Treg is mediated by HO-1 in a mouse model. HO-1 blockage by Zinc Protoporhyrin (ZnPPIX) abrogated the protective effect of Treg transfer. We found that HO-1 is important in maintaining maternal dendritic cells (DCs) in an immature state, which contributes to the expansion of the peripheral Treg population. This brings to light one essential pathway through which Treg mediates the semi-allogeneic fetus tolerance.</p> </div

    HO-1 blockage abrogates the protective effect of Treg.

    No full text
    <p>(<b>A</b>); Abortion-prone animals were adoptively transferred with Treg (2×10<sup>5</sup> cells) isolated from normal pregnant animals treatment known to prevent fetal rejection. The animals received either PBS (100 µl; n = 10) or ZnPPIX (40 mg/kg; n = 6) intraperitoneally on days 0, 3 and 6 of pregnancy and were sacrificed on day 14. (<b>B</b>)<b>:</b> As in (A) NP animals were transferred with Treg. These Treg, however, derived from either PBS- or ZnPPIX-treated donor mothers. For (A) and (B) Abortion rates were determined on day 14 of pregnancy upon animal preparation and are calculated as a percentage of the ratio of resorptions to total implantation sites. The data is presented as median abortion rates. Statistical differences were analyzed by the non-parametric Mann-Whitney-<i>U</i>- test and the statistical difference obtained between the groups was *: p<0.05 and **:p<0.01.</p

    Hmox1 and foxp3 mRNA expression positively correlate in pregnancy and HO-1 protein levels determine pregnancy outcome.

    No full text
    <p><b>A:</b> The correlation between Hmox1 and foxp3 mRNA levels in uterus of normal pregnant specimens was analyzed. The samples used in this study were uterine, decidual and placental tissues (n = 21) obtained from day 0 until 8 of pregnancy from normal pregnant (NP) females from which mRNA expression of both of <i>Hmox1</i> and <i>foxp3</i> was analyzed using real time RT PCR. Correlation analysis was performed using Pearson analysis program. r<sup>2</sup> represents co-efficient of determination value while p indicates the exact levels of significance. <b>B:</b> Total protein was obtained from decidual tissue of day 14 pregnant abortion-prone (AP) or NP animals and HO-1 levels were determined by Western Blot. The protein expression intensity level values (n = 18) were obtained from band intensities using Quantity 1 and were correlated to the abortion rates of the respective animals using Pearson's correlation analysis. The sample distribution was tested for normality using KS test. Correlation significances are indicated by p value. <b>C</b>: <i>Hmox1</i> mRNA levels in testicle samples of BALB/c (n = 3) and DBA/2J males (n = 5) were determined using real time RT-PCR. <b>D</b>: <i>foxp3</i> mRNA levels in spleen samples of BALB/c (n = 5) and DBA/2J males (n = 5) were determined using real time RT-PCR. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042301#s3" target="_blank">Results</a> are presented as median mRNA levels in dot plots. The statistical differences were analyzed using non-parametric Mann-whitney-<i>U</i>- test which gave a p value of 0.036 (*p<0.05).</p

    Splenic dendritic cells from Hmox1<sup>+/−</sup> and Hmox1<sup>−/−</sup> female mice are more mature than those obtained from Hmox1<sup>+/+</sup> controls.

    No full text
    <p>Dendritic cells (DCs) were isolated from spleen of <i>Hmox1</i><sup>+/+</sup> (n = 5), <i>Hmox1</i><sup>+/−</sup> (n = 6) or <i>Hmox1</i><sup>−/−</sup> (n = 4) females, cultured with (<b>B</b>) or without (<b>A</b>) LPS addition (1 µg/ml) and analyzed for the expression of maturity markers CD11c, CD80 and MHCII. In (<b>C</b>), the IL-10 levels secreted by DCs from these animals were measured by ELISA. Differences among the groups were analyzed by the Mann-Whitney-U test following Kruskall-Wallis test. *:p<0.05 and **:p<0.01.</p

    Dendritic cells from ZnPPIX-treated animals stimulate the proliferation of responder cells while dendritic cells from CoPPIX-treated animals influence Foxp3 expression in T cells.

    No full text
    <p>Splenic dendritic cells (DCs) from different HO-1 environment (obtained from PBS-, CoPPIX- or ZnPPIX-treated animals) were co-cultured with CFSE-stained CD4<sup>+</sup> responder cells which were isolated from lymph nodes of NP females (day 5 of pregnancy) in a ratio of 1∶1. Cells were harvested at time points 0, 24 and 48 h and proliferation was measured by flow cytometry. <b>A</b> shows the proliferation of responder cells after co-culture with DCs at time points 24 and 48 h, which was calculated with respect to time point 0. The results shown are obtained from six independent experiments. Data is shown as mean ± SEM. Statistical analyses were done using one way ANOVA. **: p<0.01. <b>B</b> shows the expression of Foxp3 in responder cells, depicted as percentage, as analyzed by flow cytometry after co-culturing these cells with DCs from animals with different HO-1 environment (24 h and 48 h). The data (mean±SEM) is representative of six independent experiments. Statistical analysis was done using one way ANOVA and # means p<0.1.</p

    In vitro CoPPIX treatment promotes IL-10 and TGF-ß secretion by dendritic cells while diminishes their ability to secrete IL-12.

    No full text
    <p><b>A–F</b> depict data obtained (mean ± SEM) in six experiments performed in duplicates or triplicates for IL-10 (pg/ml), TGF-β (pg/ml), IL-6 (pg/ml) and IL-1. In (A–D) the DCs were isolated from pregnant mice of the NO combination, while in (E–F) the DCs were obtained from animals from the AP combination. Cytokines were measured by ELISA. Statistical analysis was performed by one way ANOVA. **:p<0.01, *:p<0.05 and #:p<0.1.</p
    corecore