Th1型/Th2型细胞因子对人孕早期蜕膜的免疫调节作用

目的 :研究Th1型 /Th2型细胞因子对人孕早期蜕膜细胞活性及TGFβ1和PRL分泌的影响。方法 :蜕膜细胞活性采用MTT法进行检测 ,蜕膜组织分泌TGFβ1和PRL的活性分别采用ELISA法和RIA进行分析测定。结果 :Th1型细胞因子IFNγ低浓度时 ( 1～ 10ng/ml)对蜕膜细胞活性及PRL分泌有促进作用 (P <0 .0 5 ) ,高浓度时 ( 10 0～ 10 0 0ng/ml)则有抑制作用 (P <0 .0 1) ;IFNγ在一定剂量范围内 ( 10～ 10 0 0ng/ml)还可抑制TGFβ1的分泌 (P <0 .0 1)。而Th2型细胞因子IL 4在一定浓度范围内 ( 1～ 10ng/ml)可明显促进蜕膜的分泌活性 (P <0 .0 1)。结论 :Th1型 /Th2型细胞因子可能是通过影响蜕膜细胞活性及TGFβ1和PRL分泌而在早期妊娠中起重要的免疫调节作用     

Placental imbalance of Th1- and Th2-type cytokines in preeclampsia

OBJECTIVES: To characterize the changes in the level of T helper 1 (Th1)- [interleukin (IL)-2 and tumor necrosis factor (TNF)-alpha] and Th2-type cytokine (IL-10) and the ratios of Th1/Th2 (IL-2/IL-10 and TNF-alpha/IL-10) in placentae from women with preeclampsia and women with gestational hypertension. METHODS: Placental levels of IL-2, TNF-alpha, and IL-10 were determined with radioimmunoassay and Th1/Th2 ratios (IL-2/IL-10 and TNF-alpha/IL-10) calculated in the placentae from 22 women with preeclampsia, 15 women with gestational hypertension, and 32 normal term pregnant women. RESULTS: Although preeclampsia had the trend of the increase in the placental levels of IL-2 and TNF-alpha and the trend of the decrease in placental IL-10, there were not significant difference in placental levels of IL-2, IL-10, and TNF-alpha among preeclampsia, gestational hypertension, and normal pregnancy (P > 0.05 for all). Placental ratios of IL-2/IL-10 and TNF-alpha/IL-10 were significantly higher in preeclampsia than in normal pregnancy (P = 0.035 and P = 0.005, respectively). No differences of Th1/Th2 ratios were found between preeclampsia and gestational hypertension and between gestational hypertension and normal pregnancy (P > 0.05 for all). CONCLUSIONS: Alterations of placental balances of cytokines with Th1 predominance were demonstrated in preeclampsia. These associations may offer insights into the pathogenesis of preeclampsia.     

正常妊娠和子痫前期患者外周血树突细胞亚群的变化

目的:探讨树突细胞(DCs)及其亚群在正常妊娠和子痫前期患者间的变化,及与Th1/Th2型反应的关系。方法:选取正常妊娠孕妇25例、子痫前期患者17例和正常未孕妇女15例,用流式细胞术检测3组外周血树突细胞及其髓样(MDC)和淋巴样(PDC)亚群,比较其数量和比值在妊娠前后及子痫前期患者的变化,并与Th1/Th2型细胞因子的含量比较。结果:与正常妊娠早期和晚期相比,妊娠中期MDC和PDC数量减少,MDC/PDC比值升高,妊娠早、晚期相比无显著差异。与正常晚期妊娠妇女比较,子痫前期患者PDC数量减少,MDC数量改变不明显,MDC/PDC比值升高,两组相比差异显著。与正常晚期妊娠妇女相比较,子痫前期患者Th1型细胞因子IL-2含量增加,IFN-γ无显著差异,Th2型细胞因子IL-10减少,IL-2/IL-10、IFN-γ/IL-10比值升高。结论:DCs在正常妊娠的不同阶段其数量和亚群发生变化,子痫前期患者出现PDC减少和MDC/PDC比值升高现象,并与Th1/Th2型细胞因子的变化趋势一致。     

Thrombophilia is common in women with idiopathic pregnancy loss and is associated with late pregnancy wastage

OBJECTIVE: To describe the characteristics of thrombophilia in women with idiopathic pregnancy loss. DESIGN: Prospective observational study. SETTING: Tertiary referral center in a teaching academic hospital. PATIENT(S): One hundred forty-five patients with repeated pregnancy loss and 145 matched controls. INTERVENTION(S): Prospective assessment of thrombophilia in patients and controls. MAIN OUTCOME MEASURE(S): Prevalence of activated protein C (APC) resistance, protein C, protein S, and antithrombin III deficiencies, antiphospholipid antibodies, factor V Leiden, factor II G20210A, and MTHFR C677T mutations. RESULT(S): At least one thrombophilic defect was found in 66% of study group patients compared with 28% in control group patients. Combined thrombophilic defects were documented in 21% of women with pregnancy loss compared with 5.5% of control patients. Late pregnancy wastage occurred more frequently in women with thrombophilia compared with women without thrombophilia (160/429 [37%] vs. 39/162 [24%], respectively). APC resistance was documented in 39% of women with pregnancy loss compared with 3% of the control patients. APC resistance without factor V Leiden mutation was documented in 18% of women with pregnancy loss compared with none of the controls. While factor V Leiden mutation was more common in women with pregnancy loss (25% vs. 7.6%), factor II G20210A and homozygosity for MTHFR C677T contributed to pregnancy loss only in the presence of other thrombophilia. CONCLUSION(S): Thrombophilia was found in the majority (66%) of women with idiopathic pregnancy loss. APC resistance with or without factor V Leiden mutation is the most common thrombophilic defect, and combined thrombophilia is a frequent finding in women with pregnancy loss. Thrombophilia is associated with increased frequency of late pregnancy wastage.     

Invariant NKT cells increase lipopolysacchride-induced pregnancy loss by a mechanism involving Th1 and Th17 responses

Abstract

Objective: To determine the role of invariant natural killer T (iNKT) cells in infection-associated pregnancy loss.

Methods: B6 and iNKT cell-deficient Jα18^?/? mice were injected i.p. with lipopolysaccharide (LPS) or vehicle, and pregnancy outcomes were examined. Decidual iNKT cell expression of CD69 and intracellular cytokine production were analyzed. Mouse decidual iNKT cells were co-cultured with LPS or PBS-treated dendritic cells (DCs), and iNKT cell CD69 expression and intracellular and extracellular cytokine production were assessed.

Results: The embryo resorption rate was notably lessened for Jα18^?/? mice treated with LPS on day 6 or day 9 gestation in comparison with B6 mice treated with LPS. Decidual iNKT cell CD69 expression and intracellular IFN-γ and IL-17 production for B6 mice injected with LPS on day 6 or day 9 gestation were significantly up-regulated compared with PBS-treated mice. Levels of IFN-γ and IL-17 in the supernatants of the co-culture of decidual iNKT cells and LPS-sensitized DCs were strikingly increased in comparison with the co-culture of iNKT cells and PBS-treated DCs. CD69 expression and intracellular IFN-γ and IL-17 production of iNKT cells co-cultured with LPS-sensitized DCs were remarkably up-regulated compared with iNKT cells co-cultured with PBS-treated DCs.

Conclusions: Our results suggest that iNKT cells may play a role in LPS-induced pregnancy loss by Th1 and Th17 cytokine-dependent manner.     

The T657C polymorphism on the SYCP3 gene is associated with recurrent pregnancy loss

SYCP3 (Sinaptonemal complex protein 3) plays a critical role in pairing and recombination of homologous chromosomes in meiosis 1. It has been shown that lack of this gene leads to infertility in male and weakened fertility in female mice. In a case–control study, we investigated the SYCP3T657C polymorphism in the genome of 100 Iranian women with recurrent pregnancy losses of unknown causes as well as 100 control samples of normal fertile women having at least one healthy child. The general aim of our study was to determine whether there is a relationship between genetic changes in the SYCP3 gene and recurrent pregnancy loss in human or not. Frequency of the heterozygous genotype and mutated allele C were significantly higher in women with recurrent pregnancy losses (P-value < 0.005). Our findings suggest that the T657C polymorphism of the SYCP3 gene is possibly associated with recurrent pregnancy loss of unknown cause in human.     

Th1/Th2细胞因子在妊高征患者胎盘组织中的表达

目的 :探讨T辅助细胞 (Th) 1/Th2细胞因子在妊娠高血压综合征 (PIH)孕妇和血压正常的晚期妊娠妇女 (NLP)胎盘中的表达规律 ,从免疫学角度研究妊高征的发病原因及机制。方法 :选取PIH和NLP孕妇作为研究组和对照组 ,应用原位杂交法对两组胎盘的Th1型细胞因子 [肿瘤坏死因子 (TNF)α、白细胞介素 (IL) 2 ]和Th2型细胞因子 (IL 10 )进行标记并通过彩色病理图像分析系统对染色结果进行定量检测并作比较。结果 :(1)TNFαmRNA、IL 2mRNA在PIH及NLP组胎盘合体滋养细胞表达的平均光密度分别为0 .1978± 0 .0 32 1、0 .2 0 39± 0 .0 4 11及 0 .16 79± 0 .0 30 9、0 .16 0 0± 0 .0 4 46 (P <0 .0 0 1) ,随着PIH病情加重 ,表达逐渐增强 (P <0 .0 5 ) ;(2 )IL 10mRNA在PIH及NLP组胎盘合体滋养细胞表达的平均光密度分别为 0 .15 6 4± 0 .0 4 36及 0 .2 0 17± 0 .0 32 1(P <0 .0 0 1) ,随着PIH病情加重 ,表达逐渐减弱 (P <0 .0 5 )。结论 :在妊高征孕妇胎盘中表现为免疫杀伤的Th1型细胞因子表达增强 ,与病情呈正相关。表现为免疫保护或免疫营养的Th2型细胞因子则表达减弱 ,与病情呈负相关。提示母胎界面的Th1/Th2细胞因子平衡偏离可能是导致PIH发病的病因之一。     

Expression of CD28 and cytotoxic T lymphocyte antigen 4 at the maternal-fetal interface in women with unexplained pregnancy loss.

OBJECTIVE: To investigate the expression of CD28 and CTLA-4, two costimulatory molecules involved in T-cell activation at the maternal-fetal interface in women with unexplained pregnancy loss. METHODS: A total of 57 women, 39 with unexplained spontaneous abortion (SA) and 18 with unexplained recurrent spontaneous abortion (RSA), were enrolled in the study. A high-resolution spectratyping analysis of fluorescent bands was performed to detect CD28 and CTLA-4 expression in decidual tissues. RESULTS: The mean expression of CTLA-4 mRNA was significantly higher in women with SA than in controls (P<0.05). Moreover, the expression of CTLA-4 was higher in SA patients with genotype AA and phenotype A+ (AA+AG) than in controls (P<0.01). The expression of CTLA-4 was not significantly different in patients with RSA and in controls. The expression of CD28 was significantly decreased in patients with RSA (P<0.01) and SA (P<0.05) compared with controls. The mean ratios of CTLA-4/CD28 were significantly higher in patients with RSA (P<0.01) and SA (P<0.05) than in controls. CONCLUSIONS: This study suggests that an imbalance in CTLA-4/CD28 expression or suppressed T-cell activity at the maternal-fetal interface may confer susceptibility to unexplained pregnancy loss.     

Maternal peripheral T helper 1-type and T helper 2-type immunity in women during the first trimester of twin pregnancy

Introduction This study investigated the T helper Th1:Th2 balance in twin pregnancies compared with singleton pregnancies during the first trimester.Methods Blood samples were taken from 24 women with a singleton pregnancy and 14 women with twin pregnancy at 8–9 weeks gestation to examine the ratios of Th1:Th2 and serum human chorionic gonadotropin (hCG) and progesterone levels.Results The average ratio of Th1:Th2 in the twin pregnancies was significantly lower than that in singleton pregnancies (7.3±2.3 vs. 10.5±2.2, p<0.05). There were negative correlations between the Th1:Th2 ratio and serum hCG levels (mIU/ml) (Th1:Th2 ratio = 14.5–4.52×10^–5×hCG, r²=0.41, p<0.05) and between the Th1:Th2 ratio and serum progesterone levels (ng/dl; Th1:Th2 ratio = 23.0–0.63 × progesterone, r²=0.36, p<0.05).Conclusion Our findings show marked predominance of Th2 type cytokines occurring in twin pregnancies is related to the increase in trophoblasts during the first trimester.     

外源性细胞因子对早孕蜕膜T细胞免疫活性的研究

目的 :探讨外源性细胞因子对早孕蜕膜T细胞免疫活性作用的方式。方法 :在培养的蜕膜细胞中加入γ干扰素、白细胞介素 2、白细胞介素 6和表皮生长因子 ,共同培养 12、2 4、4 8h后收集上清液 ,并加入培养的T细胞中 ,用氚 胸苷 (3 H TdR)掺入法测定T细胞的转化值。结果 :在蜕膜与细胞因子共同培养的上清液作用下 ,各实验组T细胞的转化有不同程度的升高 ,有的T细胞转化效应显著增强 (P <0 .0 5 ) ,但增强的T细胞转化效应未显示出与作用于蜕膜的细胞因子种类、浓度及作用时间有相关性。结论 :在外源性细胞因子的作用下 ,早孕蜕膜细胞可以增强T细胞的转化作用 ,从而破坏蜕膜免疫微环境细胞因子网络的平衡状态 ,危害妊娠。     

T helper (Th)1, Th2, and Th17 interleukin pathways in infertile patients with minimal/mild endometriosis

In the present study, interleukin (IL)-10, IL-12, IL-17, and IL-23 levels were measured in serum and peritoneal fluid of women with minimal or mild endometriosis and compared with levels in controls without endometriosis. Higher IL-23 levels were encountered in the peritoneal fluid of women with endometriosis, suggesting a possible role of this cytokine in these women's infertility.     

Differential levels of T helper cytokines in preeclampsia: pregnancy, labor and puerperium.

BACKGROUND: There are suggestions that T helper 1 cytokines may be detrimental to early pregnancy and T helper 2 cytokines protective of the pregnancy. Their role in preeclamptic pregnancy, labor and puerperium, is not clear. MATERIALS AND METHODS: Twenty-eight preeclamptic women and their matched controls were evaluated, at the Departments of Obstetrics and Gynecology, Faculty of Medicine, Kuwait University and Maternity Hospital, Kuwait. Outcome measures evaluated were serum levels of TNF alpha and interleukin-4 at 12, 24, 36 weeks of gestation, advanced labor and at 1 hour and daily postpartum until they were undetectable, using ELISA technique. RESULTS: T helper cytokines showed higher serum levels in preeclampsia than normotensive pregnancy (p < 0.01, 0.01), in established labor (p < 0.05) and at 1 hour postpartum (p < 0.01 for IL-4) and p < 0.02 for TNF alpha. There was significant increase of IL-4 between 12 to 24 weeks in normal pregnancy compared to preeclampsia (p < 0.001) but not for TNF alpha. By 24 hours postpartum, IL-4 was still detectable in eight parturients compared to one patient with detectable TNF alpha (p < 0.04). Detectable IL-4 levels after 24 hours postpartum were associated with intrauterine growth retardation (p < 0.03). CONCLUSION: IL-4 has a dichotomous role in pregnancy. Normotensive pregnancy is associated with high increase in IL-4 in the first half of the pregnancy, but in the second half of pregnancy and puerperium, high levels of IL-4 are associated with preeclampsia.     

The detection of anti-beta2-glycoprotein I antibodies is associated with increased risk of pregnancy loss in women with threatened abortion in the first trimester

OBJECTIVE: This study was designed to evaluate whether the detection of serum antiphospholipid autoantibodies may be useful in predicting pregnancy outcome in women with threatened abortion in the first trimester. STUDY DESIGN: A group of 77 pregnant women of between 8 and 12 weeks' gestation with vaginal bleeding was tested for serum antiphospholipid, lupus anticoagulants, anticardiolipin, antinuclear antibodies, and anti-beta2-glycoprotein I antibodies, and was followed up until the spontaneous end of pregnancy. A control group composed of 15 healthy women with uncomplicated gestation was tested contemporarily for the same antibody panel. RESULTS: Of the 77 patients with threatened abortion, 32 (41.5%) progressed to deliver at term and 45 (58.5%) experienced early pregnancy loss. Among the antibodies evaluated, only anti-beta2-glycoprotein I was significantly more frequent in those women whose pregnancy resulted in spontaneous abortion (22/45, 49%) than in those who progressed to term (6/32, 19%) or in the control group (2/15, 13%; p=0.004). This difference was specific to the IgM isotype (p=0.001). After adjustment by multivariate analysis, the odds ratio for pregnancy loss associated with a positive beta2-glycoprotein I antibody test was 5.18 (p=0.001). CONCLUSION: The detection of anti-beta2-glycoprotein I antibodies is associated with an increased risk of pregnancy loss in women with threatened abortion in the first trimester.     

辅助性T淋巴细胞亚型与妊娠期肝内胆汁淤积症发病的关系

目的　探讨辅助性T淋巴细胞 (TH) 1型细胞因子 [γ干扰素 (IFN γ)、肿瘤坏死因子α(TNF α) ]及TH2 型细胞因子 [白细胞介素 4(IL 4) ]与妊娠期肝内胆汁淤积症 (ICP)发病的关系 ,从分子免疫学角度探讨ICP的发病机理。方法　采用双抗体夹心酶联免疫吸附法 ,检测 2 0例ICP孕妇 (研究组 )和 2 0例正常妊娠妇女 (对照组 )外周血单个核细胞在植物血凝素刺激下产生TH1型细胞因子(IFN γ ,TNF α)及TH2 型细胞因子 (IL 4)的水平 ,并对TH1型细胞因子与ICP患者血清总胆汁酸的相关性进行分析。结果　 ( 1)研究组孕妇的IFN γ、TNF α水平分别为 ( 63 9± 15 6)ng/L、( 10 2 1± 2 3 1)ng/L ;明显高于对照组孕妇的 ( 464± 147)ng/L、( 72 8± 175 )ng/L ,两组比较 ,差异有极显著性 (P <0 0 0 1)。研究组孕妇的IL 4水平为 ( 2 2± 4)ng/L ,明显低于对照组孕妇的 ( 2 7± 6)ng/L ,两组比较 ,差异有显著性(P <0 0 5 )。 ( 2 )研究组孕妇的TNF α/IL 4比值为 48± 11,明显高于对照组孕妇的 3 0± 8;研究组孕妇的IFN γ/IL 4比值为 3 0± 8,明显高于对照组的 19± 6,两组分别 ,差异均有极显著性 (P <0 0 0 1)。( 3 )研究组孕妇的IFN γ、TNF α水平与ICP患者血清胆汁酸水平呈正相关 ,相关系数 (r)分别为 0 45 8,0     

Distribution of Th1 and Th2 cell populations in human peripheral and decidual T cells from normal and anembryonic pregnancies

OBJECTIVE: To examine whether maternal immune responses during normal pregnancy are Th2 biased and whether there are specific changes when anembryonic pregnancy occurs. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology at a university hospital. PATIENT(S): We studied 32 pregnant women receiving elective abortions of normal pregnancies and 35 women with anembryonic pregnancies between 6 weeks and 10 weeks of gestational age. INTERVENTION(S): Using the multilabeling capability of three-color flow cytometry, it is possible to measure intracellular cytokines and cell surface markers simultaneously to determine which cells are the cytokine-producing cells. MAIN OUTCOME MEASURE(S): We examined the extent and proportion of mononuclear cells expressing specific T-cell surface markers and cytokines, interferon gamma, and interleukin 4 in the peripheral blood and deciduae. Secreted cytokines in the supernatants after 24-hour culture were also compared. RESULT(S): During the unstimulated status, the proportion of IL-4-secreting cells significantly exceeded that of IFN-gamma-secreting cells in the peripheral blood and decidua in normal pregnancies and was significantly decreased when anembryonic pregnancies occurred. Consequently, the Th1/Th2 ratios were increased during anembryonic pregnancies. However, after 24-hour culture, only another Th2-type cytokine, IL-10, was markedly increased and exceeded IFN-gamma secretion in cultures from both the peripheral blood and decidua in normal pregnancies. CONCLUSION(S): The decidual T lymphocytes are Th2 predominant. When anembryonic pregnancy occurs, this Th2 predominance disappears.     

The Th2 cytokines IL-4 and IL-10 are not crucial for the completion of allogeneic pregnancy in mice.

The physiological protection from fetal rejection is believed to be dependent on a Th2 type of inflammatory response at the maternal-fetal interface and the cytokines IL-4 and IL-10 have been suggested to play a critical role. We here present data from breeding experiments with IL-10 and IL-4 double-deficient mice indicating that neither maternal nor feto-placental deficiency of these cytokines are crucial for fetal or neonatal survival. The present study does not analyse possible developmental effects of maternal or fetal IL-10 and IL-4 double-deficiency in detail, but shows that an apparently normal breeding can be achieved in different crossings, providing that the mice are kept under very clean conditions.     

Th17及相关细胞因子在妊娠早期母-胎免疫耐受中的作用

目的:研究Th17细胞及相关细胞因子IL-22、IL-23在人孕早期不明原因的自发性流产母-胎免疫耐受中的作用。方法:以10例正常人工流产患者为正常早孕组,收集18例染色体正常自发性流产患者为自然流产组。分离两组患者外周血单个核细胞及蜕膜组织免疫细胞,用流式细胞术检测上述细胞中Th17细胞比例及相关细胞因子的表达,探讨其与自发性流产的关系。结果:自然流产组外周血Th17细胞比例显著高于正常早孕组(P<0.05),IL-23显著低于正常早孕组(P<0.01),但两组IL-22无显著差异;自然流产组蜕膜局部Th17细胞比例显著高于正常早孕组(P<0.01),IL-23亦显著高于正常早孕组(P<0.01),IL-22则显著低于正常早孕组(P<0.01)。结论:Th17细胞在孕早期蜕膜局部及外周血中的数量增多可能导致母胎免疫耐受失衡,在早期自然流产中发挥重要的正诱导作用,而IL-22、IL-23等细胞因子在其中的调控效应则具有明显的微环境相关性。     

Fas ligand expression by maternal decidual cells is negatively correlated with the abundance of leukocytes present at the maternal-fetal interface

The abundance of leukocytes at the maternal-fetal interface could influence the fate and invasion of extravillous trophoblasts. However, the mechanism(s) involved in determining the number of leukocytes present at the maternal-fetal interface as well as the nature of the interactions between invading fetal trophoblasts, maternal leukocytes and decidual cells are not well understood. In the present studies, we examined Fas ligand (FasL)/Fas expression at the maternal-fetal interface in human placental tissues of early pregnancy by immunohistochemistry. The types of cells and their localization were also characterized by specific cell markers (cytokeratin, vimentin and CD45 for trophoblast, decidual cells and leukocytes, respectively). The cells undergoing apoptosis and specific apoptotic trophoblasts were detected by TUNEL assay and M30 cytoDEATH immunostaining, respectively. Using single or double immunostaining, we found that FasL expression in decidual cells was negatively correlated with the number of Fas-expressing leukocytes in the same region. Furthermore, the density of leukocytes had an inverse relationship with the number of interstitial trophoblasts present in the same area. We observed also that extravillous trophoblasts are viable despite expressing Fas and being in close proximity to decidual cell-derived FasL. These data support our hypothesis that maternal decidual cell-derived FasL may be involved in preventing the recruitment of Fas-bearing leukocytes at the maternal-fetal interface through apoptosis induction by Fas/FasL interaction, thereby promoting trophoblast invasion.     

Prevalence and factors associated with pregnancy loss among physicians in King Abdul-Aziz University Hospital,Saudi Arabia

Objective

Medical profession is a stressful occupation as it carries potential risk for pregnancy outcome. There is lack of researches regarding the pregnancy loss among physicians working in hospitals in Saudi Arabia. The current study aims at estimating the prevalence and factors associated with pregnancy loss among female physicians working at King Abdul-Aziz University Hospital in Jeddah, September, 2015.