Journal of Reproductive Immunology - Articles in Press

Author's response to ‘Statins for the treatment of obstetric complications in Antiphospholipid Syndrome?’ - Corrected Proof

Guillermina Girardi — 2010-01-18

In my paper “Pravastatin prevents miscarriage in antiphospholipid antibody-treated mice” [J. Reprod. Immunol. 82 (November (2)) (2009) 126–131], I do not recommend the use of statin drugs to prevent human pregnancy loss. The first sentence of the concluding paragraph of my review article reads “We recognize that these studies were conducted in mice and that clinical trials are needed to confirm its application to humans”. I reiterate here that statin drugs should not be used in pregnant women without the evidence basis of a properly designed clinical trial to demonstrate efficacy in that setting.

Fetal cell microchimerism develops through the migration of fetus-derived cells to the maternal organs early after implantation - Corrected Proof

Rei Sunami, Mayuko Komuro, Tsutomu Yuminamochi, Kazuhiko Hoshi, Shuji Hirata — 2010-01-15

Abstract: Fetus-derived cells are present in the blood and tissues of the maternal body over a long period of time, even after delivery, resulting in fetal cell microchimerism. The exact process by which fetal cells cross the placental barrier to enter the maternal circulation is unclear. The objective of this paper was to determine the time during pregnancy that fetal cells with multilineage potential migrate to the maternal organs. Wild type female mice were crossbred with male transgenic mice, expressing enhanced green fluorescent protein (EGFP). Total hysterectomies were performed at different time points of pregnancy. On day 60 after surgery, mice were injected with either streptozotocin (STZ) to induce insulin-dependent diabetes mellitus, or vehicle. Detection and quantification of fetal cells were then undertaken in a variety of maternal organs via fluorescent microscopy and quantitative PCR amplification of the gfp transgene. In vehicle control mice, fetal cells were detected only in the maternal bone marrow. However on day 30 after STZ injection, fetal cells were detected not only in bone marrow but also in the maternal pancreas, liver and kidney. Histological analysis showed differentiated fetal cells within the pancreatic acinar cells, hepatocytes and tubular epithelial cells. Their morphological appearance was indistinguishable from their maternal counterparts, and their frequency in these organs was constant, regardless of the timing of hysterectomy. These results indicate that most fetal cells with multilineage potential in maternal tissues migrate to the maternal body early after implantation, and thereafter sustain their population over the long term after delivery.

Increased prevalence of T helper 17 (Th17) cells in peripheral blood and decidua in unexplained recurrent spontaneous abortion patients - Corrected Proof

2010-01-15

Abstract: T helper 17 (Th17) cells and regulatory T (Treg) cells are two distinct subsets of CD4+ T cells which have opposite effects on inflammation, autoimmunity and immunological rejection of foreign tissue. Treg cells have been shown to be important in maintaining materno-fetal tolerance, but the role of Th17 cells in human pregnancy and pathological pregnancy, especially in relation to unexplained recurrent spontaneous abortion (RSA), has not been investigated. In this study, we showed that the proportion of Th17 cells in the peripheral blood and decidua was significantly higher in unexplained RSA patients compared to normal, early pregnant women. Meanwhile, there was an inverse relationship between Th17 cells and Treg cells in the peripheral blood lymphocytes (PBL) and decidua in unexplained RSA. The expression of Th17 related factors, IL-17, IL-23 and retinoid orphan nuclear receptor (RORC), in PBL and decidua in unexplained RSA patients was significantly higher than normal, early pregnant women. This study is the first to define the occurrence of Th17 cells in unexplained RSA patients and in normal, early pregnant women. We suggest that these highly pro-inflammatory cells contribute to unexplained RSA, and the balance between Th17 cells and Treg cells may be critical to pregnancy outcomes.

Pro-inflammatory and anti-inflammatory cytokines in human preterm and term cervical ripening - Corrected Proof

2010-01-15

Abstract: Cervical ripening is necessary for successful delivery. Since cytokines are believed to be involved in this process, the aim of this study was to investigate possible changes in the mRNA and protein expression of pro-inflammatory cytokines (interleukin (IL)-1α, IL-1β, IL-12, IL-18) and anti-inflammatory cytokines (IL-4, IL-10, IL-13) in the human cervix during pregnancy, term and preterm labor. Cervical biopsies were taken from 59 women: 21 at preterm labor, 24 at term labor, 10 at term not in labor and 4 from non-pregnant women. mRNA was analyzed with real-time RT-PCR and protein expression and/or secretion with immunohistochemistry and ELISA. There was an upregulation of mRNA for IL-10, IL-13, IL-1α and IL-1β in the laboring groups, while mRNA for IL-12 and IL-18 was downregulated. IL-4 mRNA was detected more frequently, while IL-12 mRNA expression was lower, in the preterm labor group than in the term labor group. The protein levels of IL-4 and IL-12 were lower and IL-18 tended to be higher in the labor groups, while IL-10 protein levels were unaffected by labor. IL-4 protein levels were significantly higher in the preterm subgroup with bacterial infection than in the non-infected group. IL-10 had higher expression in squamous epithelium at preterm labor than at term. In conclusion, the major changes in pro-inflammatory and anti-inflammatory cytokine mRNA and protein expression in cervix occur during the labor process irrespective of the length of gestation. Our results indicate that dysregulation of anti-inflammatory cytokines in the human cervix could be involved in the pathogenesis of preterm labor.

Statins for the treatment of obstetric complications in antiphospholipid syndrome? - Corrected Proof

Michael D. Lockshin, Silvia S. Pierangeli — 2010-01-15

In her paper, “Pravastatin prevents miscarriages in antiphospholipid antibody-treated mice,” [J. Reprod. Immunol. 2009;82(November (2)):126–31] Dr. Girardi's conclusions go well beyond her data when she recommends the use of statin drugs to prevent human pregnancy loss.

Tumor necrosis factor-α polymorphisms in women with idiopathic recurrent miscarriage - Corrected Proof

2010-01-15

Abstract: We investigated the association of tumor necrosis factor-α (TNFα) gene polymorphisms with idiopathic recurrent miscarriage (RM). TNFα −1031T/C, −863C/A, −857C/T, −376G/A, −308G/A, −238G/A, and +488G/A single nucleotide polymorphisms (SNPs) were investigated in 204 RM women and 248 age-matched parous women by PCR-restriction fragment length polymorphism. Significantly higher frequencies of −1031C and −376A alleles were seen in RM patients; significant differences were also noted in the distribution of −1031T/C, −376G/A, and −238G/A genotypes between case and control subjects. Haploview analysis revealed high linkage disequilibrium between −857C/T and +488G/A SNPs, but was lower between the other polymorphisms. Of the possible 52 seven-locus haplotypes constructed, 10 were common, and were included in subsequent analysis. Increased frequency of CCCGGGG and CCCGGAA haplotypes, and reduced frequency of TCCGGGG and TCCGGGA haplotypes were seen in RM patients than in controls. When the Bonferroni correction was applied, differences were significant for the CCCGGAA haplotype, which was higher (OR=4.14; 95% CI=1.84–8.95), and the TCCGGGA haplotype, which were lower among RM cases (OR=0.09; 95% CI=0.02–0.68), thereby conferring RM susceptibility and protection to these haplotypes, respectively. Multivariate analysis confirmed the positive association of only CCCGGAA haplotype with RM (P=0.010; aOR=2.03; 95% CI=1.18–4.47), after controlling for a number of covariates. These results demonstrate that the TNFα polymorphisms, in particular the −1031T/C variant, are significantly associated with idiopathic RM. Additional replication studies on other racial groups are needed to confirm our findings.

Effect of vascular endothelial growth factor and interleukin-1β on apoptosis in endometrial cell cultures from patients with endometriosis and controls - Corrected Proof

2010-01-14

Abstract: The aim of this study was to evaluate the effect of vascular endothelial growth factor (VEGF) and interleukin-1β (IL-1β) on apoptosis induced by leuprolide acetate (LA) in endometrial epithelial cell cultures from patients with endometriosis. Primary endometrial epithelial cell cultures were obtained from uterine endometrial biopsies of patients with endometriosis and control women. Endometrial epithelial cells were incubated with LA; a combination of LA and VEGF; a combination of LA and IL-1β; or in basal conditions. LA was added 3h prior to addition of VEGF and IL-1β. After stimulation, the percentage of apoptotic cells was evaluated by the acridine orange–ethidium bromide technique and Bax expression was assessed by western blot. Treatment with LA enhanced the percentage of apoptotic cells in endometrial epithelial cells from subjects with endometriosis and control subjects. Addition of either VEGF or IL-1β after exposure to LA restored the percentage of apoptotic cells to basal levels. Moreover, treatment with LA increased Bax expression in endometrial epithelial cells from patients with endometriosis. This effect was reverted by the addition of either VEGF or IL-1β. Our results show that VEGF and IL-1β reduce apoptosis and decrease Bax expression in endometrial epithelial cells from patients with endometriosis. This study suggests that VEGF and IL-1β may protect endometriotic cells from undergoing apoptosis in addition to exerting their pro-angiogenic role.

Interleukin-6 and other soluble factors in peritoneal fluid and endometriomas and their relation to pain and aromatase expression - Corrected Proof

Irene Velasco, Pedro Acién, Adolfo Campos, María Isabel Acién, Eva Ruiz-Maciá — 2009-12-31

Abstract: Immunological changes and gene expression anomalies are involved in the etiopathophysiology of endometriosis, although how these alterations are connected is not well established. The aim of this study was to determine the relationship between levels of immune cell populations, cytokines and CA-125 in peritoneal fluid (PF) and ‘chocolate’ cyst fluid (CF), and aromatase expression in endometriotic tissue, as well as to investigate any association with symptoms or recurrence of the disease. Eutopic and ectopic endometrium, CF and PF were collected from 84 women with endometriomas and 24 with benign non-functioning ovarian tumors undergoing radical or conservative surgery. Immunohistochemistry was performed to determine aromatase expression. PF cell populations were assessed by flow cytometry, and CF and PF levels of interleukin (IL)-6, IL-8, IL-13, IL-17 and CA-125 were quantified by ELISA. These parameters were compared with aromatase expression, symptoms and recurrence of the disease. IL-6 levels in PF were higher in patients with endometriosis than in patients with benign non-functioning ovarian cysts, and correlated positively to dysmenorrhea and pelvic pain in the first group. An association between PF IL-8 and CA-125 was also observed in endometriosis. Aromatase positive patients showed higher levels of PF CA-125 and CF IL-17. Recurrence of symptoms or endometrioma occurred sooner in patients having higher IL-6 or IL-8 levels in CF, respectively. These findings suggest an association of IL-6 with pain in endometriosis, as well as a relationship between cytokine expression and recurrence of the disease. However no clear relationship between aromatase expression and other parameters was found.

Interleukin 11 and activin A synergise to regulate progesterone-induced but not cAMP-induced decidualization - Corrected Proof

2009-12-30

Abstract: Blastocyst implantation, placentation and the establishment of pregnancy in the human are dependent on the adequate decidualization of endometrial stromal cells. Locally produced and temporally regulated products such as interleukin 11 (IL11), and activin A are involved in this process; however, the molecular interactions that regulate decidualization are largely unknown. Here, we investigated whether IL11 and activin A interact to promote human endometrial stromal cell (HESC) decidualization using an in vitro model. HESCs, induced to decidualize by cAMP or progesterone, were treated with IL11, activin A or IL11 plus activin A combined and decidual progress was examined using prolactin as a decidual marker. Treatment with combined IL11 plus activin A enhanced progesterone-induced decidualization above control or treatment with IL11 or activin A alone. Treatment had no effect on cAMP-decidualized HESC. Investigation of IL11 and activin A stimulation of (respectively) activin A and IL11 expression in undifferentiated HESC was by real-time PCR and ELISA. Activin A treatment induced IL11 secretion and phosphorylation of the activin A signalling component, SMAD2 (measured by ELISA). Inclusion of the TGFβ 1 receptor inhibitor, SB431542, in the activin A treatment, reduced pSMAD2 and IL11 secretion. This study suggests that activin A is an early inducer of decidualization, regulating the secretion of IL11. This data provides insight into the processes underlying decidualization, which are important for understanding implantation and placentation and have potential clinical applications for the regulation of fertility.

Characterization of the invasive and inflammatory traits of oral Campylobacter rectus in a murine model of fetoplacental growth restriction and in trophoblast cultures - Corrected Proof

2009-12-30

Abstract: Campylobacter species (C. jejuni, C. fetus) are enteric abortifacient bacteria in humans and ungulates. Campylobacter rectus is a periodontal pathogen associated with human fetal exposure and adverse pregnancy outcomes including preterm delivery. Experiments in pregnant mice have demonstrated that C. rectus can translocate from a distant site of infection to the placenta to induce fetal growth restriction and impair placental development. However, placental tissues from human, small-for-gestational age deliveries have not been reported to harbor C. rectus despite evidence of maternal infection and fetal exposure by fetal IgM response. This investigation examined the temporal relationship between the placental translocation of C. rectus and the effects on fetal growth in mice. BALB/c mice were infected at gestational day E7.5 to examine placental translocation of C. rectus by immunohistology. C. rectus significantly decreased fetoplacental weight at E14.5 and at E16.5. C. rectus was detected in 63% of placentas at E14.5, but not at E16.5. In in vitro trophoblast invasion assays, C. rectus was able to effectively invade human trophoblasts (BeWo) but not murine trophoblasts (SM9-1), and showed a trend for more invasiveness than C. jejuni. C. rectus challenge significantly upregulated both mRNA and protein levels of IL-6 and TNFα in a dose-dependent manner in human trophoblasts, but did not increase cytokine expression in murine cells, suggesting a correlation between invasion and cytokine activation. In conclusion, the trophoblast-invasive trait of C. rectus that appears limited to human trophoblasts may play a role in facilitating bacterial translocation and placental inflammation during early gestation.

Stress in early pregnancy: maternal neuro-endocrine-immune responses and effects - Corrected Proof

Victoria J. Parker, Alison J. Douglas — 2009-12-28

Abstract: Stress profoundly compromises reproduction, particularly when experienced in early gestation. One outcome is pregnancy failure: although glucocorticoids have adverse effects it is not clear what their role in pregnancy failure is. However, secretion of vital hormones such as progesterone and prolactin are reduced and this unbalances the delicate and important pregnancy-protective cytokine milieu. Complex interaction between glucocorticoids, progesterone/prolactin and the immune system evidently precipitate the loss, although early loss may confer reproductive advantage by preserving maternal energy stores and facilitating ongoing maternal care for other offspring. If pregnancy failure is not induced another, perhaps more profound, outcome of maternal stress is fetal programming. Much is known about the role of elevated glucocorticoids during late gestation in fetal programming, but in early gestation their role is less clear, though likely. Other key pregnancy hormones and immune factors also contribute to fetal programming. Undoubtedly integrated action of glucocorticoids, progesterone/prolactin and the immune system is crucial for optimal pregnancy outcome and is highly susceptible to environmental conditions.

Genetic polymorphism in an inflammasome component, cervical mycoplasma detection and female infertility in women undergoing in vitro fertilization - Corrected Proof

2009-12-28

Abstract: The inflammasome is an inducible cytoplasmic structure that is responsible for production and release of biologically active interleukin-1 (IL-1). A polymorphism in the inflammasome component NALP3 has been associated with decreased IL-1 levels and increased occurrence of vaginal Candida infection. We hypothesized that this polymorphism-induced variation would influence susceptibility to infertility. DNA was obtained from 243 women who were undergoing in vitro fertilization (IVF) and tested for a length polymorphism in intron 2 of the gene coding for NALP3 (gene symbol CIAS1). At the conclusion of testing the findings were analyzed in relation to clinical parameters and IVF outcome. The frequency of the 12unit repeat allele, associated with maximal inflammasome activity, was 62.3% in cases of female infertility vs. 75.6% in cases where only the male partner had a detectable fertility problem (p=0.0095). Conversely, the frequency of the 7unit repeat allele was 28.9% in those with a female fertility problem, 17.0% in women with infertile males and 18.4% in idiopathic infertility (p=0.0124). Among the women who were cervical culture-positive for mycoplasma the frequency of the 7unit repeat was 53.7% as opposed to 19.5% in those negative for this infection (p<0.0001). We conclude that the CIAS1 7unit repeat polymorphism increases the likelihood of mycoplasma infection-associated female infertility.

The a2 isoform of vacuolar ATPase is a modulator of implantation and feto-maternal immune tolerance in early pregnancy - Corrected Proof

2009-12-16

Abstract: In mammalian reproduction, two immunologically disparate entities, the mother and her fetus, co-exist in close proximity and mutually tolerate each other. The maternal immune system plays a major contributing role in the reproductive outcome. A coordinated set of immunological events takes place between the maternal and fetal cells to ensure fetal survival. Among these, cytokines secreted by proximal maternal immune cells as well as fetal trophoblast cells play a major role in feto-maternal tolerance. In this review, we describe the role of the vacuolar ATPase (and more specifically the a2 isoform, a2V-ATPase) in controlling the expression of these vital cytokines. a2V-ATPase is a key enzyme that controls the acidification of intracellular vesicles and the extracellular environment, processes that play a major role in cellular function. The localization of a2V-ATPase in tissues and immune cells of the reproductive tract which are essential for pregnancy will be described. Information will be provided on the role of a2V-ATPase on aspects of cell development in pregnancy, from fertilization to implantation and fetal growth. Particular emphasis will be placed on the role of a2V-ATPase in (a) regulating parts of the cytokine network at the implantation site and (b) attenuating the potentially harmful maternal immune response against trophoblast cells.

Inhibition of 19S proteasomal regulatory complex subunit PSMD8 increases polyspermy during porcine fertilization in vitro - Corrected Proof

2009-11-27

Abstract: The 26S proteoasome is a multi-subunit protease specific to ubiquitinated substrate proteins. It is composed of a 20S proteasomal core with substrate degradation activity, and a 19S regulatory complex that acts in substrate recognition, deubiquitination, priming and transport to the 20S core. Inhibition of proteolytic activities associated with the sperm acrosome-borne 20S core prevents fertilization in mammals, ascidians and echinoderms. Less is known about the function of the proteasomal 19S complex during fertilization. The present study examined the role of PSMD8, an essential non-ATPase subunit of the 19S complex, in sperm-ZP penetration during porcine fertilization in vitro (IVF). Immunofluorescence localized PSMD8 to the outer acrosomal membrane, acrosomal matrix and the inner acrosomal membrane. Colloidal gold transmission electron microscopy detected PSMD8 on the surface of vesicles in the acrosomal shroud, formed as a result of zona pellucida-induced acrosomal exocytosis. Contrary to the inhibition of fertilization by blocking of the 20S core activities, fertilization and polyspermy rates were increased by adding anti-PSMD8 antibody to fertilization medium. This observation is consistent with a possible role of PSMD8 in substrate deubiquitination, a process which when blocked, may actually accelerate substrate proteolysis by the 26S proteasome. Subunit PSMD8 co-immunoprecipitated with acrosomal surface-associated spermadhesin AQN1. This association indicates that the sperm acrosome-borne proteasomes become exposed onto the sperm surface following the acrosomal exocytosis. Since immunological blocking of subunit PSMD8 increases the rate of polyspermy during porcine fertilization, the activity of the 19S complex may be a rate-limiting factor contributing to anti-polyspermy defense during porcine fertilization.

IgG is transported across the mouse yolk sac independently of FcγRIIb - Corrected Proof

2009-11-23

Abstract: While generally accepted that FcRn of the human syncytiotrophoblast and the mouse yolk sac endoderm is the major IgG transporter, the finding of a different Fc receptor FcγRIIb (RIIb) in the human placental endothelium has suggested the existence of an additional IgG transporter. Testing our hypothesis in mouse, we found that while RIIb is expressed in the yolk sac vasculature, IgG concentrations in fetuses of wild-type mice (RIIb+/+) and mice with a null mutation in the gene encoding RIIb (RIIb−/− mice) are not different, and we thus reject our hypothesis that yolk sac RIIb transports IgG in utero in the mouse. However, the capillary bed in the mouse yolk sac is structurally more complex than in human placenta, consisting of three types of cells: an RIIb-negative endothelium, a unique RIIb-bearing cell that also expresses 2 out of 4 macrophage markers but not endothelial cell or pericyte markers, and pericytes. As in the human placenta the b2 isoform of RIIb predominates in the mouse yolk sac. Remarkably only a single capillary channel rather than 2 channels with a loop is found in each yolk sac villus, which, along with intracapillary erythrocytes, suggests that blood flow is peristaltic, mediated by pericytes. It is not clear whether RIIb in the human placental villus might mediate an IgG transport function in light of the mouse yolk sac equivalent failing to do so.

WITHDRAWN: Effects of implantation and early pregnancy on the expression of cytokines and vascular surface molecules in the sheep endometrium - Corrected Proof

2009-03-23

This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

Program - Uncorrected Proof

2007-07-09