SOCS3和吲哚胺2,3-双加氧酶(IDO)在早孕绒毛和蜕膜组织中的表达呈负相关

目的探讨人早孕时细胞因子信号传送阻抑物3(SOCS3)及吲哚胺2,3-双加氧酶(IDO)在母胎界面的表达及其相关性。方法用Western blot法检测正常妊娠绒毛、蜕膜组织中SOCS3及IDO的表达。结果正常妊娠绒毛及蜕膜组织中均有IDO及SOCS3蛋白的表达,SOCS3与IDO表达呈高度负相关。SOCS3在绒毛和蜕膜组织中的表达无差异,而IDO在蜕膜组织中表达较高。结论在正常生理妊娠状态下,母胎界面中SOCS3参与调节IDO的表达,在妊娠免疫耐受的调节中起重要作用。     

IFN-γ在早孕母胎界面对吲哚胺2,3-双加氧酶(IDO)表达的正性调节研究

目的:探讨早孕绒毛和蜕膜组织中的干扰素(IFN-γ)是否上调母胎界面中吲哚胺-2,3双加氧酶(IDO)的表达。方法:收集早孕6～9周的妇女行人工流产获取绒毛组织与蜕膜组织,用Western blot方法检测绒毛、蜕膜组织中IFN-γ与IDO蛋白的表达情况;在绒毛和脱膜组织中加入不同浓度的IFN-γ(1、10和100 ng/ml)分别培养48 h和72 h后,观察IDO蛋白在两种组织中的变化。结果:人早孕绒毛和蜕膜组织均有IFN-γ与IDO蛋白的表达且成正相关性(r=0.987 1,0.979 1,P均0.01);不同浓度的IFN-γ(1、10和100 ng/ml)分别培养48、72 h后,IDO蛋白表达显著升高(P均0.05),且随着IFN-γ浓度的增高而升高。结论:在正常生理妊娠状态下,母胎界面中IFN-γ与IDO蛋白质表达成正相关,IFN-γ可上调IDO的表达。     

人早孕母胎界面SOCS1、SOCS2、SOCS3表达

目的：研究正常早孕绒毛及蜕膜组织细胞因子信号转导负调控因子（Suppressors of cytoldne signaling，SOCS）基因和蛋白水平表达，以揭示SOCS在母胎界面生理性调节作用。方法：半定量RT-PCR检测早孕绒毛组织、蜕膜组织及原代培养早孕滋养细胞、蜕膜基质细胞SOCS1、SOCS2、SOCS3 mRNA水平；Western blot检测早孕绒毛组织及蜕膜组织SOCS1、SOCS2、SOCS3蛋白表达；免疫组化定位SOCS1、SOCS2、SOCS3在早孕绒毛组织、蜕膜组织表达；ELISA检测滋养细胞、蜕膜基质细胞分泌IL-10、IFN-γ。结果：正常母胎界面见SOCS1、SOCS2、SOCS3基因表达，其中SOCS3绒毛／蜕膜阳性率73．7％／71．1％；SOCS2绒毛／蜕膜阳性率50．0％／39．5％，SOCS1最少，绒毛／蜕膜阳性率34．2％／31．6％；SOCS1、SOCS2、SOCS3蛋白表达与转录水平基本一致；正常母胎界面SOCS1、SOCS2、SOCS3表达主要定位于绒毛滋养细胞和蜕膜间质；体外无血清培养滋养细胞和蜕膜基质细胞SOCS2、SOCS3低表达，SOCS1未见表达，其分泌的IL-10随时间而增高（P〈0．05）。结论：正常早孕母胎界面表达SOCS1、SOCS2、SOCS3，无刺激条件下滋养细胞和蜕膜基质细胞低表达SOCS2、SOCS3，SOCS在正常妊娠Th平衡中具有重要意义。     

IL-6上调绒毛及蜕膜组织吲哚胺2,3-双加氧酶(IDO)的表达

目的 探讨早孕绒毛和蜕膜组织中的白细胞介素6(IL-6)对吲哚胺2,3-双加氧酶(IDO)表达的调节作用.方法 收集早孕6～9周行人工流产妇女的绒毛及蜕膜组织,采用Western blot法检测绒毛、蜕膜组织IL-6与IDO蛋白的表达情况;在绒毛、蜕膜组织中,加入(10、50、100) ng/mL IL-6分别培养48...     

复发性自然流产育龄期妇女母-胎界面中细胞因子表达分析

目的:分析复发性自然流产(RSA)育龄期妇女母-胎界面中相关细胞因子表达水平变化。方法:连续选择近期就诊的RSA育龄期妇女21例,收集流产蜕膜组织,采用酶联免疫吸附法进行蜕膜组织液中IL-2、IL-4、IL-10、IFN-γ浓度等指标检测,并与同期早孕并行人工流产终止妊娠妇女(对照组,20例)相同测试结果比较。结果:RSA组蜕膜组织液IL-2和IFN-γ浓度均明显高于对照组,而IL-4和IL-10浓度则显著低于后者(P均<0.01)。结论:RSA育龄期妇女母-胎界面上存在着明确的Th1优势状态表达。     

左旋甲基色氨酸在小鼠母胎界面诱发免疫耐受失衡

目的探讨是左旋-1-甲基色氨酸(1-L-MT)还是右旋-1-甲基色氨酸(1-D-MT)在小鼠母胎界面诱发母胎免疫耐受失衡。方法雌性BALB/c小鼠与雄性C57BL/6J小鼠交配后,自妊娠第6.5天起分别给予1-L-MT、1-D-MT、有机溶剂,共10 d,于妊娠第16.5天处死孕鼠,应用高效液相色谱法检测胎盘组织中哚胺2,3-二氧化酶(IDO)活性,流式细胞技术检测蜕膜组织中IFN-γ/IL-4比率,并对比妊娠结局。结果 1-L-MT给药组胎盘组织中IDO活性明显低于1-D-MT组及对照组,而1-D-MT组与对照组间无统计学差异;蜕膜组织中IFN-γ/IL-4比率高于1-D-MT组及对照组,而1-D-MT组与对照组间无统计学差异;胎鼠数量及体质量明显低于1-D-MT组及对照组,而1-D-MT组与对照组间无统计学差异。结论 1-L-MT而非1-D-MT通过抑制IDO活性在小鼠母胎界面诱发母胎免疫耐受失衡。     

补肾益气方通过上调滋养细胞IDO 的表达促进母胎界面NK 细胞的表型转化

目的:研究补肾益气方诱导母胎界面耐受状态的机制及吲哚胺2,3鄄双加氧酶(IDO)对蜕膜NK 细胞表型转 化的影响。方法:用免疫组织化学方法测定正常绒毛组织和复发性流产患者绒毛组织内IDO 的表达。体外培养人滋养细胞 HTR-8/ SVneo,用流式细胞术检测含药血清对HTR鄄8/ SVneo IDO 表达的影响。用含对照大鼠血清、含药大鼠血清及含药血清 联合IDO 阻断剂1鄄甲基色氨酸(1-MT)的培养液分别处理HTR鄄8/ SVneo 后与人外周NK 细胞共培养,测定其对NK 细胞的调 节作用。结果:正常绒毛组织中IDO 的表达较流产绒毛组织明显增加。含药血清可增强HTR鄄8/ SVneo IDO 的表达。外周NK 细胞与含药血清处理过的HTR鄄8/ SVneo 共培养后,NK 细胞表面CD16 表达减低,CD56 表达增加,且CD16 的改变可被1-MT 抑制。结论:补肾益气方可以通过上调滋养细胞IDO 的表达诱导外周NK 细胞向耐受型转变。     

吲哚胺2,3-双加氧酶在母胎免疫耐受中的作用研究进展

<正>作为半同种移植物的胎儿,在母体内存活直至分娩得益于母胎之间的免疫耐受。母-胎界面是胚胎和母体直接接触的部位,细胞、细胞因子及细胞外基质之间构成相互作用的复杂网路,形成了母-胎界面上特有的免疫耐受状态,以维持正常妊娠。吲哚胺2,3-双加氧酶(IDO)是分解色氨酸沿犬尿氨酸通路代谢的限速酶,通过耗竭色氨酸及产生犬尿氨酸等生物活性物质,调节T细胞、NK细胞等介导的免疫反应,发挥免疫调节作用。母-胎界面的IDO对     

早孕期人蜕膜趋化因子CCL2及其受体CCR2的表达及意义

目的：分析人早孕蜕膜及蜕膜基质细胞趋化因子受体CCR2及其配体CCL2在人早孕蜕膜组织及蜕膜基质细胞的表达和分泌，以探讨CCR2／CCL2在母-胎界面的生物学作用。方法：收集早孕期蜕膜组织，分离蜕膜基质细胞，分别用半定量RT-PCR、免疫化学方法分析正常人早孕蜕膜组织和培养的人蜕膜基质细胞CCR2／CCL2表达；并且用流式细胞术和ELISA法分别检测蜕膜基质细胞表面CCR2的表达和培养的蜕膜基质细胞上清中CCL2的分泌。结果：人早孕蜕膜组织和蜕膜基质细胞均高水平转录和翻译CCR2／CCL2，培养的基质细胞能分泌大量的CCL2，其分泌量呈时间依赖性。结论：早孕蜕膜高表达和分泌CCR2／CCL2可能参与早孕期母一胎免疫调节。     

蜕膜巨噬细胞与不明原因自然流产关系的研究进展

母胎界面多种蜕膜免疫细胞在免疫耐受中发挥作用，其中蜕膜巨噬细胞（DM）是母胎界面数量较多的免疫细胞，可分为M1型和M2型。DM向M2型极化时，机体维持正常妊娠；DM向M1型极化时，会引发自然流产（SA）。DM参与绒毛外滋养细胞侵袭、螺旋动脉重塑和免疫耐受等过程，一旦这些功能异常就会引发SA。本文对DM表型、功能及其与SA的关系进行综述。     

Reduced uterine indoleamine 2,3-dioxygenase versus increased Th1/Th2 cytokine ratios as a basis for occult and clinical pregnancy failure in mice and humans

PROBLEM: Indoleamine 2,3-dioxygenase (IDO) expression in fetal trophoblast and decidual antigen-presenting cells has been proposed to inactivate maternal T cells and thereby prevent rejection of the "fetal allograft" in early pregnancy. Psychic stress has been proposed to cause miscarriages as well as infertility, at the same time in pregnancy when blockade of IDO causes loss, but the suggested mechanism of stress-triggered loss has been an increased ratio of pro-rejection Th1-type cytokines to anti-rejection Th2/3 cytokines. Could stress act by reducing IDO expression? METHODS: Using DBA/2-mated A/J mice where stress causes early pregnancy failure, we examined the role of stress in reducing IDO versus increasing Th1/Th2 ratio in deciduas. IDO loss was also examined in human decidua associated with pregnancy failure. RESULTS: A post-implantation sonic stress increased the pregnancy failure rate, increased the Th1/Th2 ratio, but did not reduce IDO. IDO was reduced, and Th1/Th2 ratios increased in A/J mice pre-immunized against paternal DBA/2 antigens, and concomitant stress increased these effects. The rate of pregnancy failure was not further increased consistent with recent discoveries of factors that limit the impact of Th1 cytokines at the feto-maternal interface. In deciduas from spontaneous miscarriage patients, IDO(+) cell frequencies were low in only 30% of patients. CD3(+) T-cell numbers and percentage terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end-labelling (TUNEL)(+) apoptotic T cells were increased, but the level of IDO did not correlate with likelihood of apoptosis. CONCLUSIONS: Loss of an allogeneic embryo in early pregnancy is more likely to be due to a high Th1/Th2 ratio than loss of putative protection by IDO.     

The role of indoleamine‐2,3‐dioxygenase in normal and pathological pregnancies

The survival of allogeneic fetus during pregnancy contradicts the laws of immune responses. Behind this paradoxical phenomenon, the mechanism is quite complex. Indoleamine‐2,3‐dioxygenase (IDO) is the first and rate‐limiting enzyme of tryptophan catabolism. Emerging evidence shows that IDO is expressed at the maternal‐fetal interface, including trophoblast cells, decidual stroma cells, decidual immune cells (eg, natural killer cells, T cells, and macrophages), and vascular endothelial cells of decidua and chorion. Moreover, the expression and activity of IDO are different among non‐pregnant, normal pregnant, and pathological pregnant conditions. IDO plays important roles in normal pregnancy through immune suppression and regulation of fetal invasion and circulation. However, the abnormal expression and dysfunction of IDO are associated with some pathological pregnancies (including recurrent spontaneous abortion, preeclampsia, preterm labor, and fetal growth restriction).     

Global analysis of differentially expressed genes in early gestational decidua and chorionic villi using a 9600 human cDNA microarray

The global gene expression profiles of the decidua and chorionic villi of early human pregnancies were analysed by using cDNA microarray technology. Decidual and villous placental tissues were obtained from first trimester abortus and mRNA was extracted for cDNA microarray analysis. The human cDNA microarray [9600 clones, including known regulatory genes and expressed sequence tags (EST)] with colorimetric detection was used to identify differentially expressed genes between early gestational decidua and villi. According to cDNA microarray analysis, we have identified 641 genes with highly expressed mRNA in both decidua and villi, 49 genes with higher expressions in decidua, and 75 genes with higher expression in chorionic villi. These differentially expressed genes were further grouped into categories by their putative functions, including: cell growth-related factors, hormones/cytokines, cell adhesion molecules, signal transduction molecules, apoptosis-related factors, cytoskeleton/extracellular matrix proteins, and EST. Immunohistochemical stainings of cathepsin L, leukaemia inhibitory factor-receptor, and proliferative cell nuclear antigen showed results consistent with the microarray data. Identification of the differentially expressed genes between decidua and villi by microarray provide a global profiling of the gene expression pattern. This work adds to our understanding of placentation by reporting the gene expression profiles during first trimester human pregnancies using cDNA microarray.     

Phenotypic characterization of regulatory T cells in the human decidua

Pregnancy is a unique situation for the maternal immune system. We have studied and identified a CD4+CD25+ regulatory T (Treg) cell population isolated from the human decidua. This mucosal surface in the uterus is in direct contact with semiallogenic fetal cells. We observed that about 14% of the decidual CD4+ T cells have the CD4+CD25+ phenotype. The decidual CD4+CD25+ T cells expressed high frequency of intracellular CTLA-4 (CTLA-4i). The majority of CD4+CD25+CTLA-4i+ cells were also positive for GITR and OX40, typical markers for human Treg cells. The frequency of CD4+CD25+ T cells in the peripheral blood from pregnant women was found to be increased during the first and second trimester of gestation when compared to nonpregnant controls. Being an important molecule for Treg cells, the role of CTLA-4 in the regulation of indoleamine 2,3-dioxygenase (IDO) expression was also examined. The stimulation with CTLA-4Ig did not increase IDO mRNA expression in CD14+ cells from pregnant women, while IFN-gamma was observed to up-regulate IDO expression. The presence of Treg cells in the human decidua suggests that these cells are important in protecting the fetus from alloreactive immune responses at the maternal-fetal interface.     

HLA-DR positive cells in the human placenta.

A population of heterogeneous HLA-DR positive cells has been identified in the human placenta and decidua using immunochemical and histochemical methods. These cells are found in three areas: the subepithelial layer of the amnion, the decidua, and more sparsely within the chorionic villous stroma. In addition to HLA-A, -B and -C antigens, they also express the leucocyte-common antigen, indicating their origin from bone marrow precursors. The majority have a characteristic stellate shape with many cytoplasmic processes. In the villous stroma these stellate cells can be distinguished from the Hofbauer cells (placental macrophages) by their morphology, stronger expression of HLA-DR and lack of lysosomal enzyme activity. In the amnion and decidua they cannot be clearly distinguished from tissue macrophages. By using monoclonal antibodies specific for foetal or maternal HLA-A or -B allotypes, the HLA-DR positive cells in the chorionic villi and the amnion have been shown to be foetal in origin. In contrast, most of the HLA-DR positive cells in the decidua are maternal; a few adjacent to the basal plate are foetal. The preponderance of these cells in those areas of the placenta where foetal and maternal tissues are in close proximity is striking. The possibility that some of the cells are equivalent to the dendritic cells that have been described in other tissues is discussed.     

Prenatal diagnosis of glycogenosis type II (Pompe''s disease) using chorionic villi biopsy

Glycogenosis type II (Pompe's disease) has been diagnosed using cultured amniotic cells for several years. In this paper, we present three prenatal diagnoses based on chorionic villi biopsy in three families at risk for Pompe's disease juvenile form: a normal fetus that was diagnosed and confirmed by enzymatic assay on amniotic cells; two affected fetuses that were diagnosed and confirmed on post-abortion fetal tissues. In one case a residual acid alpha-glucosidase activity was found; we concluded that the residual activity was due to maternal contamination. Prenatal diagnosis of Pompe's disease is therefore possible using chorionic villi biopsy.     

Analysis of HLA-DRB1*-A* and -B* alleles in prenatal diagnosis for determination of maternal contamination in fetal DNA

During chorionic villi sampling for prenatal diagnosis with molecular biology techniques, contamination by maternal decidua frequently occurs and can lead to misinterpretation of the test results. To avoid such problems, we present a new method for appraising maternal contamination of fetal DNA, based on genomic typing of the highly variable human leukocyte antigen (HLA) locus-DRB1*, locus A* and locus B* regions by genetic amplification with sequence-specific primers and PCR. Fetal DNA samples obtained for beta-thalassemia diagnosis were analysed after artificial contamination with increasing maternal DNA concentrations ranging from 0.5 to 10% (0.5, 1, 3, 5 and 10%). The approach was found to be rapid, specific, reproducible and highly sensitive and permits recognition of 1-3% contamination by maternal DNA concentrations. The system currently used for detecting maternal DNA contamination in fetal samples is the analysis of polymorphic loci by variable number of tandem repeats and/or short tandem repeats. We propose that the analysis of HLA alleles may provide a valid alternative or complement to this system.