Showing papers in "American Journal of Reproductive Immunology in 2012"

Immunomodulatory Properties of Mesenchymal Stem Cells: Cytokines and Factors

Abstract: Mesenchymal stem cells (MSCs) are defined as undifferentiated cells that are capable of self renewal and differentiation into several cell types such as chondrocyte, adipocyte, osteocyte, myocyte, hepatocyte, and neuron-like cells. MSC can be isolated from bone marrow, umbilical cord blood, adipose tissue, placenta, periosteum, trabecular bone, synovium, skeletal muscle, and deciduous teeth. Immunomodulatory of MSCs is one of the important issues nowadays, because this aspect can be clinically applied for graft-versus-host and autoimmune diseases. In this review, we tried to discuss in detail about cytokines and factors such as members of the transforming growth factor superfamily (transforming growth factor-β), hepatic growth factors (HGF), prostaglandin E2 (PGE2), IL-10, indolamine 2,3-dioxygenase (IDO), nitric oxide (NO), heme oxygenase-1 (HO-1), and human leukocyte antigen-G (HLA-G) that are involved in immunomodulatory of MSCs.

Models of fetal brain injury, intrauterine inflammation, and preterm birth.

Abstract: Intrauterine infection and inflammation are known risk factors for brain damage in the neonate irrespective of the gestational age. Infection-induced maternal immune activation leads to a fetal inflammatory response mediated by cytokines that has been implicated in the development of not only periventricular leukomalacia and cerebral palsy but also a spectrum of neurodevelopmental disorders such as autism and schizophrenia (Behav Brain Res 2009; 204:313, Ann Neurol 2005; 57:67, Am J Obstet Gynecol 2000; 182:675). A common link among the neurobehavioral disorders associated with intrauterine inflammation appears to be the evidence for immune dysregulation in the developing brain (Behav Brain Res 2009; 204:313). The timing of the immune challenge with respect to the gestational age and neurologic development of the fetus may be crucial in the elicited response (J Neurosci 2006; 26:4752). Studies involving animal models of maternal inflammation serve a key role in elucidation of mechanisms involved in fetal brain injury associated with exposure to the maternal milieu. These animal models have been shown to result in fetal microglial activation, neurotoxicity as well motor deficits and behavioral abnormalities in the offspring (J Neurosci 2006; 26:4752, J Neurosci Res 2010; 88:172, Am J Obstet Gynecol 2009; 201:279, Am J Obstet Gynecol 2008; 199:651). A better understanding of the mechanisms of perinatal brain injury will allow discoveries of novel neuroprotective agents, better outcomes following preterm birth and stratification of fetuses and neonates for therapies in cases of preterm birth, preterm premature rupture of membranes, and chorioamnionitis.

Th17 and regulatory T cells in women with recurrent pregnancy loss.

Abstract: The immune system of pregnant women is tightly controlled to defend against microbial infections and at the same time, to accept an embryo or the fetus, which are expressing semi-allogenic paternal antigens. Furthermore, inflammation-like processes are crucial for tissue growth, remodeling, and differentiation of the decidua during pregnancy. Dysregulation of elaborate immune control may lead reproductive failure, such as implantation failure, recurrent pregnancy loss (RPL), preterm birth, intrauterine fetal growth restriction, and preeclampsia. Until recent years, a balance between Th1 and Th2 cells was believed to be the key immune regulatory mechanism of T-cell immunology especially during pregnancy. Since the identification of regulatory T cells was made, the mechanism of immune regulation has become a major issue in immunologic research. Also, the recent identification of Th17 cells has drawn our attention to a new immune effector. The balance between Th17 and regulatory T cells may explain more about the pathophysiology of reproductive failure. This review will discuss relevant human literature on regulatory T and Th17 cells in normal reproductive physiology and in women with RPL and infertility.

Short‐Chain Fatty Acids Induce Pro‐Inflammatory Cytokine Production Alone and in Combination with Toll‐Like Receptor Ligands

Abstract: Problem Short chain fatty acids (SCFAs), produced at relatively high levels by anaerobic bacteria in bacterial vaginosis (BV), are believed to be anti-inflammatory. BV, a common alteration of the genital microbiota associated with increased susceptibility to HIV infection, is characterized by increased levels of both pro-inflammatory cytokines and SCFAs. We investigated how SCFAs alone or together with TLR-ligands affected pro-inflammatory cytokine secretion.

Does Immunotherapy for Treatment of Reproductive Failure Enhance Live Births

Abstract: Before effective treatment for reproductive failure can be instituted, the cause of the failure must be determined. A search of PubMed was made to identify the published data regarding diagnosis and treatment of reproductive failure. Results were compared with the frequency of antiphospholipid antibodies (APA) in 2995 women with histories of unexplained infertility, recurrent implantation failure, recurrent pregnancy loss, and fertile women. In addition, pregnancy outcomes among 442 women experiencing reproductive failure and elevated NK cell activity after treatment with intravenous immunoglobulin (IVIg) (N = 242) or intralipids (N = 200) were compared. The prevalence of APA was the same among women with the diagnosis of unexplained infertility, recurrent implantation failure, and recurrent miscarriage. Heparin and aspirin are successful in the treatment of elevated APA among women with recurrent miscarriage but not with recurrent implantation failure. IVIg has been successful in the treatment of recurrent miscarriage and recurrent implantation failure among women with elevated APA and/or NK cell activity. When the pregnancy outcomes of women with a history of reproductive failure and elevated NK cell cytotoxicity treated with intralipid were compared with women treated with IVIg, no differences were seen. Immunotherapy for treatment of reproductive failure enhances live birth but only in those women displaying abnormal immunologic risk factors.

Modulation and recruitment of inducible regulatory T cells by first trimester trophoblast cells.

Abstract: Citation Ramhorst R, Fraccaroli L, Aldo P, Alvero AB, Cardenas I, Leiros CP, Mor G. Modulation and recruitment of inducible regulatory T cells by first trimester trophoblast cells. Am J Reprod Immunol 2012; 67: 17–27 Problem The specialized regulatory T-cells (Treg) population, essential for maternal tolerance of the fetus, performs its suppressive actions in the critical peri-implantation phase of pregnancy. In the present work, we investigated whether trophoblast cells are able to induce Treg recruitment, differentiation, and whether these mechanisms are modified by a bacterial or viral infection. Method of Study Human T-regulatory cells were differentiated from naive CD45RA+ CCR7+ cells obtained from peripheral blood mononuclear cells cultured with IL-2 and TGFβ over 5 days. Induction of iTregs (CD4+ Foxp3+ cells) was evaluated using low serum conditioned media (LSCM), obtained from two first trimester trophoblast cell lines, Swan-71 and HTR8. Coculture experiments were carried out using transwell assays where trophoblast cells were in the absence or presence of PGN, LPS, or Poly [I:C]. Cytokine production was measured by multiplex analysis. Results Trophoblast cells constitutively secrete high levels of TGFβ and induced a significant increase of Foxp3 expression accompanied by a specific T-reg cytokine profile. Moreover, trophoblast cells were able to recruit iTregs in a specific manner. Conclusion We demonstrate that trophoblast cells have an active role on the recruitment and differentiation of iTregs, therefore, contributing to the process of immune regulation at the placental–maternal interface.

Intravenous immunoglobulin treatment increased live birth rate in a Spanish cohort of women with recurrent reproductive failure and expanded CD56(+) cells.

Abstract: cells, in vitro fertilization, intravenousimmunoglobulin, NK cells, recurrentmiscarriageCorrespondenceSilvia Sa´nchez-Ramo´n, Division of ClinicalImmunology, Department of Immunology,Hospital General Universitario GregorioMaran˜o´n, Calle Doctor Esquerdo 46, 28007Madrid, Spain.E-mail: ssanchez.hgugm@salud.madrid.orgSubmission November 20, 2011;accepted March 13, 2012.CitationMoraru M, Carbone J, Alecsandru D, Castillo-Rama M, Garci´a-Segovia A, Gil J, Alonso B,Aguaro´n A, Ramos-Medina R, Marti´nez deMari´a J, Oliver-Min˜arro D, Rodri´guez-MahouM, Ortega V, Caballero P, Meliā E, Vidal J,Cianchetta-Sivori M, Esteban C, Vargas-HennyL, Dale J, Ortiz-Quintana L, Ferna´ndez-Cruz E,Sa´nchez-Ramo´n S. Intravenousimmunoglobulin treatment increased live birthrate in a Spanish cohort of women withrecurrent reproductive failure and expandedCD56

The frequency of peripheral blood CD4+ CD25high FoxP3+ and CD4+ CD25- FoxP3+ regulatory T cells in normal pregnancy and pre-eclampsia.

Abstract: Problem Regulatory T cells (Tregs) play an important role in the development of pregnancy-specific immune tolerance. We aimed to determine the peripheral frequency of a recently described Treg subpopulation, the CD4+ CD25− FoxP3+ Treg subset, and its correlation with the conventional CD4+ CD25high FoxP3+ Tregs in normal pregnancy (NP) and pre-eclampsia (PE) compared to non-pregnant (non-P) women. We also examined the proportion of the activated CD4+ CD25high FoxP3high Treg subset within conventional Treg cells. Method We took peripheral blood samples from 20 PE, 20 NP, and 12 non-P women and determined the frequency of the above Treg subsets using flow cytometry. Results The frequency of conventional CD4+ CD25high FoxP3+ Tregs and activated CD4+ CD25high FoxP3high Tregs, but also that of non-conventional CD4+ CD25− FoxP3+ Tregs was higher in NP compared to non-P women, but lower again in PE, reaching comparable levels to the non-P group. The ratios of CD4+ CD25high FoxP3+ and CD4+ CD25− FoxP3+ Treg subsets were constant in all three investigated groups. Conclusion Our results indicate that the frequency of conventional and non-conventional Tregs alters simultaneously, and the presence in circulation of both of these Treg subsets is similarly important in the adequate development of pregnancy-specific immune tolerance.

Methylome of Fetal and Maternal Monocytes and Macrophages at the Feto‐Maternal Interface

Abstract: Problem Decidual macrophages (dMϕ) of the mother and placental macrophages (Hofbauer cells, HC) of the fetus are deployed at a critical location: the feto-maternal interface. This study was conducted to compare DNA methylome of maternal and fetal monocytes, dMϕ, and HC, and thereby to determine the immunobiological importance of DNA methylation in pregnancy.

Biomarkers of Coagulation, Inflammation, and Angiogenesis are Independently Associated with Preeclampsia

Abstract: Biomarkers of coagulation, inflammation and angiogenesis are independently associated with preeclampsia

Changes of NK Cells in Preeclampsia

Abstract: The regulation of uterine and circulating peripheral blood natural killer (NK) cells has been associated with reproductive immunology such as recurrent pregnancy losses, implantation failures, or preeclampsia. Preeclampsia is a hypertensive disorder of pregnancy characterized by increased blood pressure accompanied by proteinuria and is a major cause of maternal and fetal mortality. Natural cytotoxicity receptors (NCRs) are unique markers, which regulate NK cell cytotoxicity and cytokine production. The relation of NCRs to reproduction is not fully characterized yet. The different profile of NCRs expression may suggest presence of abnormal regulation of NK cell in women with reproductive failures. Pregnant women with preeclampsia carry immunological abnormalities of NCRs on peripheral blood NK cells during pregnancy. The lower expression of NKp46(+) NK cells in women with preeclampsia may account for the higher production of NK1 cytokine that is known as NK1 shift in pregnant women with preeclampsia. Evaluation of NKp46 on peripheral blood NK cells may be applicable to find the onset of preeclampsia. In this review, various expressions of NK cell surface markers including NCRs on NK cells, NK cell cytotoxicity, and production of cytokines and angiogenic factors by NK cells were reviewed in relation to preeclampsia.

Explants of Intact Endometrium to Model Bovine Innate Immunity and Inflammation Ex Vivo

Abstract: Problem Bacterial infections commonly cause bovine endometritis and infertility via innate immune pathways. However, mechanistic studies using isolated cells or chopped tissue may be compromised by the disruption of endometrial architecture and release of damage-associated molecular patterns. So, this study aimed to establish an ex vivo model of intact bovine endometrium to study innate immunity and inflammation. Method of study Intact bovine endometrium explants were collected using a sterile 8-mm punch biopsy and cultured ex vivo with bacteria or pathogen-associated molecules. Interleukin accumulation was measured, and tissue viability was assessed by microscopy, TdT-mediated biotin–dUTP nick-end labelling and lactate dehydrogenase assay. Results Intact endometrium explants accumulated IL-6, IL-1β and IL-8 in response to Gram-negative or Gram-positive bacteria, and their purified pathogen-associated molecules; inflammatory responses were dependent on the stage of oestrous cycle. Explants of intact endometrium maintained viability and tissue architecture, and had lower basal accumulation of interleukins compared with explants using chopped endometrium. Conclusion This study established a tractable ex vivo model of intact endometrium to explore the mechanisms of immunity and inflammation in the bovine endometrium.

The balance of the immune system between T cells and NK cells in miscarriage.

Abstract: Immunological dysfunction has been proposed to explain the etiology of recurrent pregnancy loss (RPL). The immunological environment differs between the decidua basalis and decidua parietalis, and also between RPL cases with normal fetal chromosomes and those with abnormal fetal chromosomes. The problem with analyzing decidual tissues from spontaneous abortions is that cause versus effect phenomena are difficult to distinguish. Recent data show that the immune system in a late-stage miscarriage is completely different from that in an early-stage miscarriage. If immunocompetent cells can cause RPL, the immunological environment may be a causative factor, especially in an early-stage miscarriage, at the decidua basalis, and/or in cases of RPL with a normal embryo. Careful examination of the immune system at the decidua basalis in an early-stage miscarriage in RPL cases with normal fetal chromosomes may reveal useful information. This paper aimed at finding a cause of RPL by analyzing the balance of the immune system between T cells and NK cells in an early-stage miscarriage.

An increase of Treg cells in the peripheral blood is associated with a better in vitro fertilization treatment outcome.

Abstract: Problem The objective of this study was to determine whether there was any association between the peripheral blood CD4+ CD25+ Foxp3+ regulatory T cells (Treg cells) and implantation success in patients undergoing in vitro fertilization (IVF) treatment. Method of study Prospective observational study of 101 randomly selected women who underwent IVF treatment for tubal factor from May 2011 to June 2011. The percentage of peripheral blood Treg cells and the expression levels of Foxp3 and CTLA4 mRNA in peripheral blood mononuclear cells (PBMCs) were recorded and their relations to IVF treatment outcomes were analyzed. Results Treg cells were significantly elevated in the pregnant group (P = 0.03). The expression level of Foxp3 mRNA in PBMCs from pregnant group also significantly increased (P = 0.02). A receiver operating characteristic analysis (area under curve = 0.631) found that those women with Treg cells >0.6%, the pregnancy rate and live birth rate were much higher as compared to women with Treg cells below this level (P < 0.05). An increase of Treg cells in the peripheral blood was associated with a better IVF treatment outcome (OR 4.3, 95% CI = 1.76–10.48), with a sensitivity of 64%, specificity of 71%. Conclusion An elevated level of circulating Treg cells was associated with increased rates of pregnancy and live birth in IVF treatment.

Genetic Polymorphisms and Recurrent Spontaneous Abortions: An Overview of Current Knowledge

Abstract: The relevance of gene polymorphisms in the development of unexplained recurrent spontaneous abortion is still unclear. Cytokines, angiogenic mediators, and hormones are involved in all stages of reproduction and pregnancy outcome. Impaired production and/or unbalanced ratios of these mediators have been implicated in the pathogenesis of unexplained recurrent spontaneous abortion. Functional polymorphism influence gene activity and therefore can interfere with the expression of mediators. Several studies have been carried out to evaluate the relationship between cytokines, angiogenic mediators, and hormones gene polymorphisms and unexplained recurrent spontaneous abortion. The results of these studies are mostly contradictory, and few significant associations have been identified. Up to present time, the evidence is insufficient to support the evaluation of cytokines, angiogenic mediators, and hormones gene polymorphism in routine workup in all cases of recurrent pregnancy loss, and these tests are not included in any of the major obstetric guidelines.

CD4+CD25highFoxp3+ Cells Increased in the Peritoneal Fluid of Patients with Endometriosis

Abstract: PROBLEM To evaluate CD4(+) CD25(high) Foxp3(+) cells and IL-6, IL-10, IL-17, and TGF-β in the peritoneal fluid of women with endometriosis. METHOD OF STUDY A total of ninety-eight patients were studied: endometriosis (n = 70) and control (n = 28). First, peritoneal fluid lymphocytes were isolated, and CD4(+) CD25(high) cells were identified using flow cytometry. Then, RT-PCR was performed to verify Foxp3 expression in these cells. Also, IL-6, IL-10, IL-17, and TGF-β concentration was determined. RESULTS Of all the lymphocytes in the peritoneal fluid of women with endometriosis, 36.5% (median) were CD4(+) CD25(high) compared to only 1.15% (median) in the control group (P < 0.001). Foxp3 expression was similarly elevated in patients with the disease compared to those without (median, 50 versus 5; P < 0.001). IL-6 and TGF-β were higher in endometriosis group (IL-6: 327.5 pg/mL versus 195.5 pg/mL; TGF-β: 340 pg/mL versus 171.5 pg/mL; both P < 0.001). IL-10 and IL-17 showed no significant differences between the two groups. CONCLUSION The peritoneal fluid of patients with endometriosis had a higher percentage of CD4(+) CD25(high) Foxp3(+) cells and also higher levels of IL-6 and TGF-β compared to women without the disease. These findings suggest that CD4(+) CD25(high) Foxp3(+) cells may play a role in the pathogenesis of endometriosis.

Multiple Pregnancy Failures: An Immunological Paradigm

Abstract: Recurrent spontaneous abortion (RSA), three or more pregnancy losses prior to 20 weeks, occurs in about 1% of all pregnancies, 50% of RSA cases remain unexplained and unresolved. Recently, immune pathways have been implicated in the pathophysiology of RSA. Immune tolerance of the fetal-placental unit and placental angiogenesis are mandatory for a successful pregnancy outcome. Unscheduled dysregulation of the placental vasculature is thought to be the pathophysiologic mechanisms underlying an array of pregnancy complications like infertility, miscarriage, pre-eclampsia, and fetal growth restriction and death. Investigations on mechanisms and management of RSA are mired by substandard design and lack of optimal randomized clinical trials and have resulted in disagreement on guidelines for evaluation and treatments for patients with multiple pregnancy losses of unknown etiology. The present review focuses on evidence-based research discussion with immunologic causes, and immune-regulatory therapies recommended for helping patients with a history of RSA. We highlight data that might support revalidation of low molecular weight heparin as a protective therapy in RSA. Newly launched growth factors, GM-CSF, and potentially novel agents to suppress inflammatory rejection, including regulatory T cells, human chorionic gonadotropin, and M-CSF/IL-10, may work in concert with tender-loving-care therapy and give hope to couples with multiple pregnancy losses.

A subset of human uterine endometrial macrophages is alternatively activated.

Abstract: Problem Human uterine macrophages must maintain an environment hospitable to implantation and pregnancy and simultaneously provide protection against pathogens. Although macrophages comprise a significant portion of leukocytes within the uterine endometrium, the activation profile and functional response of these cells to endotoxin are unknown. Method of Study Flow cytometric analysis of surface receptors and intracellular markers expressed by macrophages isolated from human endometria was performed. Uterine macrophages were stimulated with LPS. Cytokines, chemokines, and growth factors expressed by these cells were analyzed using Bio-Plex analysis. Results CD163high human endometrial macrophages constitutively secrete both pro- and anti-inflammatory cytokines as well as pro-angiogenic factors and secretion of these factors is LPS-inducible. Conclusion A major population of human uterine macrophages is alternatively activated. These cells secrete factors in response to LPS that are involved in the activation of immune responses and tissue homeostasis.

A Cross‐Talk of Decidual Stromal Cells, Trophoblast, and Immune Cells: A Prerequisite for the Success of Pregnancy

Abstract: Embryo implantation and formation of a functional placenta are complex processes that require a plethora of regulatory mechanisms involving both mother and embryo cells. Recently, an important role in this complicated cells and factors network was assigned to the decidual stromal cells (DSC) and trophoblast cells. Decidualization includes biochemical changes that trigger DSC to produce a number of factors required for the implantation and induction of immunotolerance in maternal immune system. Immunotolerance is achieved by a cascade of strictly controlled events starting with selective homing of immune cells to the feto-maternal site, regulated proliferation, and predominant differentiation into a regulatory type of immune cells. Furthermore, cytotoxic effector functions are reduced owing to the influence of steroid hormones, factors, cytokines, and inhibitory receptors. Altogether the entire immune system of the mother is switched to tolerogenic functional state which is a prerequisite for the successful maintenance of pregnancy.

The Treg/Th17 imbalance in Toxoplasma gondii-infected pregnant mice.

Abstract: Citation Zhang H, Hu X, Liu X, Zhang R, Fu Q, Xu X. The Treg/Th17 Imbalance in Toxoplasma gondii-Infected Pregnant Mice. Am J Reprod Immunol 2012; 67: 112–121 Aim To evaluate whether impaired Treg/Th17 balance exists in the pregnant mice infected with Toxoplasma gondii. Method of Study Regulatory T (Treg) and T-helper type 17 (Th17) cells were measured in both placenta and spleens of the pregnant mice infected with and without T. gondii by flow cytometry. The mRNA and protein expression levels of transforming growth factor-β (TGF-β) and interleukin-17A (IL-17A) were analyzed using real-time PCR and enzyme-linked immunosorbent assay (ELISA). The mRNA expression levels of forkhead box P3 (Foxp3), retinoic acid–related orphan receptor γt (RORγt), and IL-6 were also analyzed using real-time PCR. The correlations of the ratio of Treg/Th17 to the mRNA or protein expression level of those factors were analyzed by Spearman’s correlation analysis. Data were analyzed by unpaired t-test and paired t-test. Results The proportion of Tregs or Th17 cells in the placenta and spleens of the T. gondii-infected pregnant mice was significantly lower or higher than in those of non-infected mice, respectively. Upregulation of TGF-β and downregulation of IL-17A were found in the placenta of T. gondii-infected pregnant mice. The ratio of Treg to Th17 was significantly lower in the infected mice than that in the non-infected mice (P < 0.01).The ratio of Treg to Th17 positively or negatively correlated with the protein expression level of TGF-β (r = 0.6204, P < 0.05) or IL-17A (r = −0.6296, P < 0.05), respectively. The ratio also positively correlated with the mRNA expression level of Foxp3 (r = 0.7985, P < 0.01), but negatively correlated with the mRNA expression level of RORγt (r = −0.6153, P < 0.05), and IL-6 (r = −0.7492, P < 0.01). Conclusion TheTreg/Th17 imbalance exists in the pregnant mice infected with T. gondii, which is associated with the expression of related cytokine and key transcription factors. This result suggests that the embryo loss caused by this parasite may be associated with a reduced ratio of Treg to Th17 cell number.

Imbalance in Cytokines from Interleukin‐1 Family – Role in Pathogenesis of Endometriosis

Abstract: Problem To assess whether interleukin (IL)-1beta, IL-18 and interleukin-1 converting enzyme (ICE) are involved in the pathogenesis of endometriosis. Method of study Peritoneal fluid (PF) was obtained from 85 women with and without endometriosis. Peritoneal macrophages were cultured and the culture media collected. IL-1beta, IL-18 and ICE levels were measured by the enzyme-linked immunosorbent assay (ELISA). Results Levels of IL-1beta and ICE in PF of women with endometriosis were higher than those in the control group. However, PF level of IL-18 was significantly lower in the study group than in the controls. Higher secretion of IL-1beta by peritoneal macrophages and lower IL-18 and ICE in endometriosis patients than in control were observed. Following lipopolysaccharide (LPS) stimulation, the macrophages secreted more IL-1beta, IL-18 and ICE in all groups. Conclusions The results pointed to impairment of the secretion of the IL-1 cytokine family in endometriosis. Invalid IL-1beta and IL-18 maturation by ICE may be an important pathogenic factor in endometriosis.

Expression of Kisspeptin and its receptor GPR54 in the first trimester trophoblast of women with recurrent pregnancy loss.

Abstract: Citation Park D-W, Lee S-K, Hong SR, Han A-R, Kwak-Kim J, Yang KM. Expression of kisspeptin and its receptor GPR54 in the first trimester trophoblast of women with recurrent pregnancy loss (RPL). Am J Reprod Immunol 2012; 67: 132–139 Problem Kisspeptin and its receptor GPR54 play a major role in trophoblast invasion. The expression of kisspeptin and GPR54 in trophoblast and decidua and their relationship with decidual and peripheral blood natural killer (NK) cells are investigated in women with RPL. Method of study Trophoblast and decidual tissues were collected from 38 RPL women who miscarried a genetically normal fetus and 14 women who had elective abortion. Kisspeptin, GPR54, and decidual NK cells were investigated with immunohistochemistry, and peripheral blood NK cells were analyzed by flow cytometry. Results Kisspeptin expression in syncytiotrophoblast was significantly decreased in RPL women with normal (<15%) peripheral blood NK cells (npNK) (P = 0.021) and high (≥15%) peripheral blood NK cells (hpNK) (P = 0.024) as compared to controls. Kisspeptin expression in cytotrophoblast was significantly decreased hpNK group (P = 0.009) as compared to controls. GPR54 expressions were not different among study groups and controls. The number of CD56+ decidual NK cells are significantly higher in hpNK group as compared to npNK group (P = 0.041) and showed a correlation with kisspeptin expression in syncytiotrophoblasts (r = 0.738, P < 0.001). Conclusion Decreased kisspeptin expression in trophoblasts is associated with RPL and kisspeptin may engage the regulation of decidual NK cell infiltration.

Detailed Analysis of Peripheral Blood Natural Killer Cells in Women with Repeated IVF Failure

Abstract: Problem To analyse the peripheral blood NK cells in women with repeated IVF failure (RIF) and a fertile control group to determine which parameters best differentiate the two populations. Methods Peripheral blood from the luteal phase of 171 women with RIF and 33 fertile controls was analysed by four-colour flow cytometry for NK cell concentration, subset differentiation and the activation marker CD69. Results Women with RIF had significantly increased NK cell numbers as determined by concentration (P < 0.05) and percentage of lymphocytes (P < 0.001), increased concentration of the CD56dim subtype (P < 0.05), and increased concentration of activated CD56dimCD69+ cells (P = 0.0001). There was no correlation between any NK cell parameters with the length of infertility or number of embryo transfer cycles. Conclusions Peripheral blood NK cell activity is significantly higher in women with RIF than in fertile controls. Future trials of immune therapy in women undergoing IVF should target those with high NK activity.

Lipoxin A4 Inhibits the Development of Endometriosis in Mice: The Role of Anti‐Inflammation and Anti‐Angiogenesis

Abstract: Problem To evaluate the effects of the anti-inflammatory and anti-angiogenic roles of LXA4 on endometriosis in mice. Method of study Endometriosis was induced in 40 mice and separated into two groups. LXA4 group was administered by LXA4 for 3 weeks. The endometriotic lesions were counted, measured, and identified by pathology. The presence of a panel of pro-inflammatory factors was assessed by real-time RT-PCR, and enzyme-linked immunoassay, the mRNA, protein levels of matrix metalloproteinase (MMPs), and vascular endothelial growth factor (VEGF) were determined by real-time RT-PCR and immunohistochemistry; the activity of MMPs was evaluated by gelatin zymography. Results Treatment with LXA4 significantly inhibited endometriotic lesion development (13.58 ± 4.01 mm2 in LXA4 group and 23.20 ± 7.49 mm2, P = 0.0002), downregulated pro-inflammatory factors, suppressed the activity of MMP9, and reduced the VEGF levels associated with endometriosis in mice. Conclusion LXA4 may inhibit the progression of endometriosis possibly by anti-inflammation and anti-angiogenesis.

Glucocorticoid receptor mediates the effect of progesterone on uterine natural killer cells.

Abstract: Problem Uterine natural killer cells (uNK) do not express progesterone receptor, but express glucocorticoid receptor (GR). So, we speculate that progesterone may regulate uNK cells through a GR-mediated process. Method of Study After mouse NK cells were stimulated with CpG with or without IL-12 in the presence or absence pre-treatment of progesterone, the effects of progesterone on NK via GR were investigated by using RU486 (progesterone receptor and GR antagonist) and CDB-2914 (progesterone receptor antagonist). The expressions of CD69 and IFN-γ were determined by flow cytometry and qPCR. Phosphorylation of IκB and STAT4 was determined by Western blot. Furthermore, we purified uNK cells from human decidual tissues using anti-CD56 microbeads to verify the effect of progesterone on uNK via GR. Results Progesterone suppressed CD69 and IFN-γ expression of mouse spleen NK cells and human uNK cells induced by CpG combined with IL-12. CDB-2914 had no effect on IFN-γ expression suppressed by progesterone, while RU486 could abolish the inhibitory effect of progesterone. In addition, progesterone could decrease the phosphorylation of both STAT4 and IκB. Conclusions In the present study, we first prove that progesterone can regulate NK cells via GR. It is valuable for further understanding the role of uNK in progesterone regulated gestation process.

Postpartum Group A Streptococcus Sepsis and Maternal Immunology

Abstract: Group A Streptococcus (GAS) is an historically important agent of puerperal infections and sepsis. The inception of hand-washing and improved hospital hygiene drastically reduced the incidence of puerperal sepsis, but recently the incidence and severity of postpartum GAS infections has been rising for uncertain reasons. Several epidemiological, host, and microbial factors contribute to the risk for GAS infection and mortality in postpartum women. These include the mode of delivery (vaginal versus cesarean section), the location where labor and delivery occurred, exposure to GAS carriers, the altered immune status associated with pregnancy, the genetic background of the host, the virulence of the infecting GAS strain, and highly specialized immune responses associated with female reproductive tract tissues and organs. This review will discuss the complicated factors that contribute to the increased susceptibility to GAS after delivery and potential reasons for the recent increase observed in morbidity and mortality.

Distinct subpopulations of epithelial ovarian cancer cells can differentially induce macrophages and T regulatory cells toward a pro-tumor phenotype.

Abstract: Problem The presence of immune infiltrates in the tumor does not always correlate with an anti-tumoral immune response. We previously identified two sub-populations of epithelial ovarian cancer (EOC) cells with differential cytokine profile. We hypothesize that these two sub-populations of EOC cells may differentially regulate the immune phenotype in the tumor microenvironment and therefore affect the immune response.

Peripheral CD300a+CD8+ T Lymphocytes with a Distinct Cytotoxic Molecular Signature Increase in Pregnant Women with Chronic Chorioamnionitis

Abstract: Citation Xu Y, Tarquini F, Romero R, Kim CJ, Tarca AL, Bhatti G, Lee J, Sundell IB, Mittal P, Kusanovic JP, Hassan SS, Kim J-S. Peripheral CD300a+CD8+ T lymphocytes with a distinct cytotoxic molecular signature increase in pregnant women with chronic chorioamnionitis. Am J Reprod Immunol 2012; 67: 184–197 Problem CD300a is an immunomodulatory molecule of the immunoglobulin receptor superfamily expressed in the leukocytes of myeloid and lymphoid lineages. However, its biological function on CD8+ T lymphocytes remains largely unknown. This study was conducted to assess the biological significance of CD300a expression in T lymphocytes and to determine whether its expression in peripheral T lymphocytes changes in pregnant women presenting with antifetal rejection. Methods of Study Microarray analysis was performed using total RNA isolated from peripheral CD300a+ and CD300a− T lymphocytes. Flow cytometric analysis of the peripheral blood samples of pregnant women and pathologic examination of the placentas were conducted. Results A large number of genes (N = 1245) were differentially expressed between CD300a− and CD300a+ subsets of CD8+ T lymphocytes, which included CCR7, CD244, CX3CR1, GLNY, GZMB, GZMK, IL15, ITGB1, KLRG1, PRF1, and SLAMF7. Gene ontology analysis of differentially expressed genes demonstrated enrichment of biological processes such as immune response, cell death, and signal transduction. CD300a expression in CD8+ T lymphocytes was coupled to a more cytotoxic molecular signature. Of note, the proportion of CD300a+CD8+ T lymphocytes increased in pregnant women with chronic chorioamnionitis (antifetal rejection of the chorioamniotic membranes; P < 0.05). Conclusion The findings of this study strongly suggest an increase in systemic T-lymphocyte-mediated cytotoxicity in pregnant women with chronic chorioamnionitis as a manifestation of maternal antifetal rejection.

Immune etiology of recurrent pregnancy loss and its diagnosis.

Abstract: Recurrent Spontaneous Abortion of Immunological Origin (RSAI) is currently diagnosed by the occurrence of 2–3 consecutive miscarriages of unknown origin. The psychological trauma incurred by these events is a serious ailment which may be potentially avoided if a method of analysis is derived which may forecast these events and in turn prevent them from occurring. This review intends to examine studies of recurrent spontaneous abortion (RSA) which use laboratory diagnosis and also studies of RSA that do not use laboratory diagnosis. We believe that when laboratory results are incorporated into the diagnosis of RSA/RSAI that treatment is highly successful whereas the absence of laboratory results severely hinders the effectiveness of treatment. It is worth noting that correlating treatment versus outcome is imprudent because of the multiple variables involved in patient cases. It is not imprudent, however, to say that incorporation of laboratory data is essential when diagnosing RSA/RSAI.

Profile of inflammatory mediators in gestational diabetes mellitus: phenotype and genotype.

Abstract: Citation Gueuvoghlanian-Silva BY, Torloni MR, Mattar R, de Oliveira LS, Scomparini FB, Nakamura MU, Daher S. Profile of inflammatory mediators in gestational diabetes mellitus: phenotype and genotype. Am J Reprod Immunol 2012; 67: 241–250 Problem Our study aimed to assess in vitro production of IL-10, IL-6, TNF-A, and adiponectin serum levels in pregnant women with and without gestational diabetes mellitus (GDM) and to investigate a possible association between GDM and IL-10−1082 A>G (rs1800896), IL-6−174 G>C (rs1800795), TNF-A−308 G>A (rs1800629), adiponectin +45 T>G (rs2241766), and adiponectin−11377 C>G (rs266729) gene polymorphisms. Method of study This case–control study included 79 women with GDM and 169 healthy controls (C) grouped according to pre-pregnancy BMI. IL-10, IL-6, and TNF-A culture supernatant and adiponectin serum levels were assessed by ELISA. DNA genotype was performed by PCR-RFLP. Results Adiponectin levels were significantly higher in C than GDM women, even within the same BMI category. Cytokines levels were similar between the groups. There were no associations between GDM and the analyzed gene polymorphisms. Conclusions Women with GDM have significantly lower adiponectin levels in the third trimester, regardless of BMI.