早期妊娠的免疫耐受机制

胚胎对于母体来说是半异己的移植物 ,其表达部分父系抗原。孕早期母体免疫系统 ,尤其是子宫内膜局部的 NK细胞、巨噬细胞、TCR( γ+ δ+ ) T细胞和 B淋巴细胞等免疫细胞能否识别并耐受胚胎抗原 ,直接影响妊娠结局。近年来的研究表明 ,母体免疫系统在受到体内高浓度性激素水平抑制的基础上 ,通过子宫内膜局部 EAIF、HLA-G等分子产生免疫抑制 ;R80 K、BP和 PIBF等封闭因子产生免疫封闭 ;Fas L和 RCAS1诱导免疫清除 ;降调 CD1、CD80、CD86及增强 IL-1 0表达以诱导克隆无能和 /或非特异性的免疫调节等机制共同作用 ,产生对胚胎抗原的免疫耐受 ,而且这些因素之间存在相互作用 ,其中一种机制失调即引致母体对胚胎的免疫排斥 ,可导致妊娠失败     

Altered dendritic cell function in normal pregnancy

In pregnancy, maternal immunity is skewed to favour maintenance of gestation and immune tolerance of a semi-allogeneic fetus. Dendritic cells are thought to play a crucial role in mediating the balance between immunity and tolerance, and determining the type of T helper cell response. We postulated that myeloid dendritic cells would be modified in pregnancy to favour type 2 T helper cell responses. We show that the proportion of circulating myeloid dendritic cells expressing CD86 and staining for HLA-DR were significantly lower in the third trimester of pregnancy compared with non-pregnant women. As pregnancy progressed through the third trimester to term, CD86 expression increased. Furthermore, monocytes from pregnant women differentiated into less phenotypically mature dendritic cells which expressed lower levels of CD80, CD86 and HLA-DR molecules compared with non-pregnant women. In response to inflammatory stimuli, monocyte-derived dendritic cells, from pregnant women up-regulated CD86 more than CD80, and secreted less IL-12p70 but more IL-10, compared with monocyte-derived dendritic cells from non-pregnant controls. Our results demonstrate that, in pregnancy, the dendritic cell system is modified to favour type 2 T helper cell responses.     

白介素β1及凝血酶降低孕早期蜕膜细胞同源框A10基因的表达

目的:探讨炎症及出血导致流产的作用机制。方法:选择复发性流产(RSA)及正常妊娠孕6~10周的蜕膜组织各10例,并在体外分离培养正常妊娠的蜕膜细胞,加入雌、孕激素、白介素β1(IL-β1)及凝血酶处理后,用实时荧光定量RT-PCR及Western blotting检测RSA患者、正常妊娠者蜕膜组织及培养处理的各组蜕膜细胞内的同源框A10(HOXA10)的表达情况。结果:1HOX A10基因在孕早期蜕膜细胞中有表达;与正常妊娠者相比,RAS患者蜕膜组织HOXA10的表达明显下降(P0.05)。2雌、孕激素处理组的蜕膜细胞HOXA10 mRNA的表达显著增加(P0.05),是未处理组的9.5倍。3进一步加入IL-β1或凝血酶后,HOXA10 mRNA表达显著下降(P0.05);与雌、孕激素组相比,分别下降89.5%和74.9%。结论:1 HOXA10基因在孕早期蜕膜细胞中有显著表达,在RAS患者中的表达显著下降。2雌、孕激素促进HOXA10的表达,而IL-β1及凝血酶抑制HOXA10的表达。     

Activation antigens during the proliferative and secretory phases of endometrium and early-pregnancy decidua

BACKGROUND: Clarifying the normal distribution of activation antigens will contribute to database construction studies of monoclonal-antibody-based therapies in endometrial disorders. METHODS: In this study, endometrial tissue samples obtained during proliferative and secretory phases and decidual samples of early pregnancies were immunostained by the monoclonal antibodies anti-CD26, anti-CD30, anti-CD70, anti-CD71, and anti-CD98 using the indirect immunoperoxidase method. RESULTS: CD26 is expressed on the glandular epithelium in the endometrium and decidua. Endothelial CD26 is expressed less in the decidua when compared to the endometrium. CD30 is strongly expressed by decidual cells. It is only weakly expressed on endometrial and decidual vessels. Glandular and endothelial CD70 expression is mainly seen in the proliferative phase of the menstrual cycle. Glandular CD71 expression is less in the decidua when compared to the endometrium. Its expression on stromal cells is more in the secretory phase of the menstrual cycle and in early pregnancy deciduae. It is expressed on endometrial vessels but not on decidual vessels. Glandular CD98 is expressed more in the decidua when compared to the endometrium. This antigen exists on endometrial lymphocytes. It is strongly expressed on the endothelium in the endometrium and decidua. CONCLUSION: It seems that CD26 and CD70 are not involved in the functions of endometrial and decidual stromal cells. CD30 and CD71 are thought to be involved in decidualization. Absence of activation antigens other than CD98 on lymphocytes indicated an antigenic profile for large granular lymphocytes that is different from regular lymphocytes.     

补肾益气和血方中药对胚泡着床障碍小鼠子宫内膜表面胞饮突表达的影响

目的　观察补肾益气和血方中药对胚泡着床障碍小鼠子宫内膜表面胞饮突表达的影响。方法　选择 8～ 12周龄昆明小鼠 ,于妊娠第 4天 (Pd4 )晨 ,皮下注射米非司酮 ,制成胚泡着床障碍模型。然后将妊娠小鼠随机分为正常组、模型组和治疗组 ,治疗组从Pd1开始灌服中药 (补肾益气和血方 ) ,正常组和模型组则给予等量生理盐水灌服 ,模型组和治疗组于Pd4晨皮下注射米非司酮 ,正常组则注射等量生理盐水。采用扫描电镜观察各组小鼠在Pd4晚 2 1时 30分～ 2 2时 (第 1时间点 ,T1)和Pd5上午 9时 30分～ 10时 (第 2时间点 ,T2 ) ,共两个时间点子宫内膜表面胞饮突的表达。结果　模型组妊娠率 ( 2 5 % )较正常组 ( 90 % )显著降低 ,差异有显著性 (P <0 0 5 ) ,模型组平均着床胚泡数为 ( 7 7± 1 3)个 ,正常组为 ( 14 7± 1 4 )个 ,两组比较 ,差异有极显著性 (P <0 0 0 1) ;治疗组的妊娠率 ( 5 5 % )及平均着床胚泡数 [( 12 3± 0 8)个 ]均较模型组显著增加 (P <0 0 5 ,P <0 0 1)。T1时正常组子宫内膜表面表达大量发育中的胞饮突 ,至T2时则表达大量发育完全的胞饮突 ;T1时模型组子宫内膜发育不同步 ,胞饮突仅在局部少量表达 ,至T2时则表达完全消失 ;T1时治疗组子宫内膜表面表达大量发育中的胞饮突 ,与正常组相比略延     

Immunosuppressive effect of progesterone on dendritic cells in mice

Progesterone has been demonstrated to be involved in maintaining pregnancy by regulating immunocytes. Dendritic cells (DCs), the most potent triggers of the adaptive immune response, express receptors for steroid hormones and are regarded as one of the primary targets of progesterone. However, the functional modification of DCs by progesterone remains poorly understood. Here, we report that progesterone does not affect the morphology or apoptosis of murine bone marrow-derived DCs. Progesterone-treated DCs were characterized by decreased expression of Ia (MHC class II), CD80 and CD86, increased production of IL-10, and decreased secretion of IL-12. Compared with mature DCs (mDCs), activated progesterone-treated DCs had a reduced capacity to stimulate CD4(+) T cell proliferation. The observation that progesterone-treated DCs could attenuate delayed-type hypersensitivity (DTH) responses in vivo suggests that progesterone mediates suppressive DC activity. However, transfer of progesterone-treated DCs into the peritoneal cavity of mice did not elevate the percentage of CD4(+)CD25(+)Foxp3(+) regulatory T cells in the spleen. Overall, our study helps to increase understanding of the role of DCs exposed to progesterone in the maintenance of pregnancy.     

妊娠早期阻断协同刺激分子诱导自然流产模型孕鼠脾脏细胞母-胎免疫耐受的研究

目的　探讨阻断协同刺激分子———CD80 和CD86对自然流产模型孕鼠妊娠结局及孕鼠脾脏免疫细胞对父系抗原免疫耐受状态的影响。方法　将雌性小鼠 (CBA/J)分别与BALB/c及DBA/2两种雄性小鼠合笼交配 ,分别建立正常妊娠模型CBA/J×BALB/c( 2 0只 ,对照组 )和自然流产模型CBA/J×DBA/2 ( 2 0只 ,研究组 )。CBA/J小鼠于妊娠第 4天 (着床期 )腹腔分别注射大鼠同型IgG 0 2mg( 10只 ) ,或大鼠抗小鼠CD80 和CD86单克隆抗体 ( 10只 )。妊娠第 9天 ,采用单向混合淋巴细胞反应 ,分析孕鼠脾脏免疫细胞对父系抗原的增殖能力 ,并测定细胞培养上清液中白细胞介素 2(IL 2 )水平 ,以研究脾脏细胞母 胎免疫耐受状态 ;妊娠第 14天观察两组的胚胎吸收率。结果　 ( 1)研究组中 ,腹腔注射大鼠IgG的孕鼠胚胎吸收率为 2 4 3% ,而注射大鼠抗小鼠CD80 和CD86单克隆抗体的孕鼠胚胎吸收率为 9 8% ,两者比较 ,差异有显著性 (P <0 0 5 )。 ( 2 )应用大鼠抗小鼠CD80 和CD86单克隆抗体 ,使妊娠 9d的孕鼠脾脏免疫细胞对父系抗原的增殖能力及IL 2水平显著下降(P <0 0 5 )。结论　孕早期阻断协同刺激分子 ,可诱导产生孕鼠脾脏免疫细胞对父系抗原的免疫耐受 ,从而使自然流产模型孕鼠的妊娠结局达到正常妊娠水平。     

Uptake of 3H-norepinephrine in different segments of the human non-pregnant and pregnant uterus

Tissue specimens from uterine cervix and corpus of non-pregnant and pregnant women were incubated in vitro in the presence of 3H-norepinephrine. The uptake in the cervix exceeded those of the lower and upper segments of the corpus, and this difference was most pronounced at term pregnancy. The neuronal uptake in the cervix constituted approximately 80% of the total uptake and was higher than in isthmus and fundus (50 and 20%, respectively). The results favour the notion that cervical nerves remain functionally intact throughout pregnancy and support previous histochemical studies demonstrating a segmental difference in uterine innervation and a partial denervation of the myometrium at term.     

IL-10在子宫腺肌病种植窗期在位内膜的表达与子宫腺肌病内膜容受性的关系

目的：分析IL-10在子宫腺肌病患者在位内膜种植窗期的表达,探讨子宫腺肌病患者内膜容受性下降的免疫学因素。方法：分别取子宫腺肌病、非腺肌病患者在自然周期种植窗期的内膜组织。子宫腺肌病患者18例,对照组16例,采用免疫组化SP法检测内膜组织中IL-10的表达。结果：IL-10在腺肌病患者内膜腺体和间质的表达较非腺肌病患者减弱（P〈0．05）,差异有统计学意义。结论：子宫腺肌病患者在位内膜中具有妊娠保护作用的IL-10表达减低,推测IL-10的表达降低是导致子宫腺肌病患者内膜容受性下降的免疫学因素。     

不同条件下小鼠围着床期子宫内膜整合素α_v和Le~y寡糖的表达

目的:探讨促排卵药、胚胎本身及孕后激素等条件对小鼠子宫内膜整合素av和Ley寡糖的表达及对着床窗开放的影响。方法:将成年雌鼠分为对照和实验两大组。对照组为自然受孕;实验组为用促排卵药后受孕,其中一部分为假孕,另一部分为事先行一侧输卵管结扎。各组分别于孕2-7 d取子宫标本2份,一份石蜡包埋行HE染色和免疫组化染色测定Ley寡糖,另一份于-80℃保存,冰冻切片行免疫组化染色测定整合素av。结果:①对照组于孕7 d,大部分小鼠子宫外观即见明显胚胎着床,显微镜下从孕5 d起出现子宫内膜间质的蜕膜反应,但实验组无1例有此变化。②整合素av在小鼠子宫内膜腺上皮细胞胞浆内表达。对照组的表达于孕4 d达高峰,而各实验组的表达高峰均推迟1-2 d,且表达水平较对照组明显减弱(P <0.05)。结扎侧和未结扎侧子宫内膜整合素av的表达高峰无明显差异。③Ley寡糖在小鼠着床期表达于子宫内膜腔上皮和腺上皮表面,其表达两组间无明显差异,不同妊娠天数之间的表达亦无明显差异。在对照组,于孕4 d开始Ley寡糖在子宫内膜间质有表达,孕6 d达高峰,而实验组未见一定的规律。结论:①小鼠子宫内膜腺上皮细胞胞浆内整合素av的表达与子宫内膜着床窗的开放一致。②促排卵药能抑制小鼠子宫内膜着床窗期整合素av的表达,延迟着床窗的开放;孕     

雌、孕激素对人子宫内膜Pbx2蛋白表达的影响

目的:探讨人子宫内膜腺上皮细胞和基质细胞中雌、孕激素对Pbx2蛋白表达的影响。方法:采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(SP)连接法检测人增生期、分泌中期和蜕膜期子宫内膜中Pbx2的表达和分布;体外培养人子宫内膜基质细胞和高分化子宫内膜癌细胞Ishikawa,分别加入雌激素、孕激素、雌、孕激素联合刺激48 h,并以不加雌、孕激素的基质细胞核和Ishikawa细胞为对照组,采用免疫组织化学、免疫印迹法测定各种条件下Ishikawa细胞中Pbx2蛋白的表达。结果:①在人各期子宫内膜组织中,Pbx2表达于腺上皮细胞和基质细胞的细胞核中;Pbx2在分泌中期和蜕膜期基质细胞中的表达显著高于增生期,但在腺上皮细胞中增生期组织的表达高于分泌中期和蜕膜期,差异均具有统计学意义(P<0.05)。②Western blotting结果显示,在孕激素和雌、孕激素联合处理Ishikawa细胞组中,Pbx2的表达显著低于对照组(P<0.05),雌激素处理后Pbx2的表达与其他各组间的表达无显著性差异(P>0.05),在人子宫内膜基质细胞(ESC)中,Pbx2的表达在雌、孕激素处理各组间比较均无统计学差异(P>0.05)。结论:在人子宫内膜腺上皮Ishikawa细胞中孕激素对Pbx2的表达具有下调作用,在人子宫内膜基质细胞中,Pbx2的调控可能是非雌、孕激素依赖性的。     

子宫性索样肿瘤2例及文献分析

目的 报告2例子宫性索样肿瘤（USCT）,探讨其临床病理特征与生物学行为。方法 观察2例USCT的临床表现、发生部位、光镜形态与免疫表型。结果 2例肿瘤分别发生于宫体与宫颈。肿瘤上皮样细胞呈巢状、索状、梁状等性索状排列,部分细胞胞质泡沫状。免疫组化2例均表达CK（广谱）、CD99、SMA、Vimentin、CD10,1例表达CD117。结论 USCT可发生于宫体与宫颈,光镜与免疫表型兼具上皮、性索、平滑肌与子宫内膜间质的部分特征,生物学行为有待更多病例的积累。     

Trophoblast debris modulates the expression of immune proteins in macrophages: a key to maternal tolerance of the fetal allograft?

Interactions between maternal immune cells and the placenta are of substantial interest since diseases of pregnancy, such as recurrent miscarriage, villitis of unknown etiology and preeclampsia may arise due to inadequate adaptation of the maternal immune system. During normal pregnancy trophoblast debris is shed from the placenta into the maternal blood in large quantities. This trophoblast debris is then rapidly cleared from the maternal circulation. In this study, we exposed trophoblast debris generated from an in vitro placental explant model to peripheral blood-derived macrophages and quantified a variety of molecules that are important in immune responses by ELISA or flow cytometry. Phagocytosis of trophoblast debris resulted in reduced cell-surface expression of MHC-II molecules, the costimulatory molecules (CD80, CD86, CD40 and B7H3), monocyte chemoattractant protein-1 (MCP-1), inter-cellular adhesion molecule 1 (ICAM-1) and IL-8 receptors in macrophages while the expression of programmed death-1 ligand 1 (PD-L1) was upregulated. In addition, phagocytosis of trophoblast debris induced the secretion of the anti-inflammatory cytokines IL-10, IL6 and IL1Ra and decreased the secretion of pro-inflammatory cytokines IL-1β, IL12p70 and IL-8 by macrophages. Phagocytosis of trophoblast debris also increased macrophage expression of the immunosuppressive enzyme indoleamine 2,3-dioxygenase (IDO). We have shown that phagocytosis of trophoblast debris from normal placentae alters the phenotype of macrophages such that they are likely to deviate maternal immune responses towards tolerance and away from inflammation. This may be one of the mechanisms that allow the human fetal allograft to survive in direct contact with the maternal immune system.     

大剂量米非司酮和孕激素联合应用治疗子宫内膜癌的临床研究

目的　研究大剂量米非司酮、大剂量孕激素以及二者联合应用对子宫内膜癌患者的治疗效果 ,并探讨其作用机制。方法　将 3 0例经诊刮确诊的子宫内膜癌患者随机分为 3组 ,每组 10例 ,术前用药 5d。米非司酮组术前用米非司酮 ( 10 0mg d) ,醋酸甲羟孕酮组术前用醋酸甲羟孕酮 ( 50 0mg d) ,联合组术前用米非司酮 ( 10 0mg d)、醋酸甲羟孕酮 ( 50 0mg d)。各组用药前后行自身对照 ,观察癌细胞组织形态学改变及雌激素受体 (ER)、孕激素受体 (PR)、增殖细胞核抗原 (PCNA)、凋亡相关基因 (bcl 2、bax)及CD44 v6基因表达的变化。结果　各组治疗后 ,癌细胞分化均趋向成熟 ,分泌活跃 ,可在癌组织中观察到凋亡的癌细胞 ,联合组治疗后变化最明显。醋酸甲羟孕酮组治疗前后 ,免疫组织化学 (免疫组化 )评分分别为PR( 2 9± 1 1、1 6± 0 8)、ER( 2 8± 0 9、1 4± 0 9)、PCNA( 0 84± 0 11、0 60±0 12 )、bcl 2 ( 0 2 3 6± 0 0 89、0 157± 0 981)和CD44 v6( 4 6± 1 8、2 5± 1 9) ,表达均降低 (所有P <0 0 1) ,bax( 0 2 0± 0 10、0 42± 0 0 7)表达增多 (P <0 0 1)。米非司酮组治疗前后免疫组化评分分别为PR( 3 4± 1 0、1 9± 0 8)、ER( 2 7± 0 9、1 2± 0 7)、PCNA( 0 80± 0 15、0     

Effect of medroxyprogesterone acetate on serum levels of LH, FSH, cortisol, and estrone in patients with endometrial carcinoma

Medroxyprogesterone acetate (MPA) was administered orally for at least 6 months to 25 patients with endometrial adenocarcinoma starting 1 week after hysterectomy and bilateral ovariectomy. We measured serum concentrations of MPA during treatment and serum levels of LH, FSH, cortisol, and estrone before and during treatment with MPA. The serum levels of MPA 3--5 h after ingestion of a daily 100-mg dose of MPA at 08.00 h, varied considerably, the mean value (+/- SE) being 43 +/- 3.6 nmol/l range 15--119 nmol/l. Serum FSH levels rose after operation in the five premenopausal patients and remained elevated despite MPA treatment. In the 20 postmenopausal patients, however, initially high serum FSH and LH levels declined to half the starting values as a result of MPA treatment. After the administration of MPA for 6 months the mean serum cortisol and estrone concentrations at 08.00 a.m. had fallen to 78% and 82% of the mean values before treatment.     

Endocrinology of the peri-implantation period.

Current research suggests that the appearance of endometrial integrins and pinopode appearance signal the opening of the receptive phase of the endometrium. These integrins may be activated by the interleukin-1 system (IL-1). IL-1beta, expressed by the blastocyst, induces vascular endothelial growth factor (VEGF) which, in turn, promotes angiogenesis and integrin expression in endometrial cells. The IL-1 system also triggers the expression of gamma interferon (IFN-gamma) from T lymphocytes. Decidual natural killer (NK) lymphocytes interact with invading trophoblast to generate leukaemia inhibitory factor (LIF). LIF induces uPA and gelatinase, enzymes which play a crucial role in trophoblastic invasion.Progesterone is a potent inhibitor of LIF, while oestrogen is a potent inducer. Oestrogen in serum reflects follicular IL-1beta level and correlates with the outcome of embryo transfer after in vitro fertilization (IVF). Progesterone induces nitric oxide (NO) synthesis in the decidua, and NO promotes local vasodilatation and uterine quiescenceMeasurement of placental protein 14 (PP14, glycodelin-A) in serum may be of value as a screening test for implantation potential. However, human chorionic gonadotrophin (hCG) remains the most reliable predictor of successful implantation and pregnancy viability. An ovulation + 14 hCG level < 50 IU/l is often predictive of a non-viable outcome, while an ovulation + 21 hCG of < 200 IU/l always indicates a non-viable pregnancy. hCG secretion by invading trophoblast appears to be negatively modulated by endothelin-1 (ET-1) and prostaglandin F(2alpha)(PGF2alpha), while tissue growth factors and collagenases are positive modulators of hCG expression.ProalphaC, an inhibin pro-monomer, may have some value in monitoring corpus luteum function. Inhibin A, activin A and follistatin all rises throughout pregnancy and peak at 36 weeks of gestation. Relaxin is another ovarian hormone that may have a role in predicting implantation. Relaxin induces placental protein 14 (PP14, glycodelin-A) expression in a receptive endometrium, and measurement of serum PP14 may be of value as a screening test for implantation potential.     

Endometrial expression and secretion of interleukin-6 throughout the menstrual cycle.

Previous findings revealing reduced endometrial interleukin-6 (IL-6) mRNA expression in patients with recurrent early abortions gave rise to the analysis of IL-6 synthesis in the human endometrium, and how it relates to concentrations in uterine secretions. Endometrial tissues and uterine secretions were collected from patients undergoing hysterectomy. Expression of IL-6 receptor and gp130 mRNA (n = 28) in total endometrium, of IL-6 mRNA in endometrial epithelial and stromal cells, and CD45-positive leukocytes were investigated by RNAase protection assay throughout the cycle. IL-6 protein was assessed in endometrial tissue by immunohistochemistry (n = 32) and in uterine secretions by enzyme-linked immunosorbent assay (ELISA) (n = 33). IL-6 mRNA was expressed at low levels in the proliferative phase, and expression increased progressively in the secretory phase. The increase was attributed to epithelial and stromal cells and leukocytes. Concentrations of IL-6 protein in endometrial glands and in uterine secretions were low in the proliferative phase, and increased 5-10-fold in the mid- to late secretory phase. mRNA expression of IL-6 receptor and gp130 remained constant in total endometrium throughout the menstrual cycle. High concentrations of IL-6 in the mid-secretory phase, the putative implantation window, and a further increase in the late secretory phase, the premenstrual period, support a role of IL-6 in the regulation of endometrial functions.     

Pregnancy associated hormones modulate the cytokine production but not the phenotype of PBMC-derived human dendritic cells

OBJECTIVE: Dendritic cells (DC) play a central role in initiating and polarizing immune responses. As effects of pregnancy associated hormones on phenotype and function of DC are unknown, our objective was to test the influence of progesterone, beta-estradiol and betaHCG on immature (iDC) and mature (mDC) DC. STUDY DESIGN: DC generated from peripheral-blood-monocytes were exposed to different doses of hormones. DC phenotype was determined by FACS-analysis of surface marker expression (CD40, CD86, CD83 and HLA-DR). Modifications in the secretion of cytokines (IL12p70, IL-18, IL-10, IL-6, TNFalpha) and chemokines (MDC, IL-8) were analysed by ELISA. T cell stimulatory capacity of mDC was assessed by mixed lymphocyte reaction. RESULTS: Incubation with progesterone or estradiol resulted in a significant upregulation of IL-10 production by iDC and mDC. Combinations of progesterone and betaHCG or estradiol respectively induced a significant decrease in production of IL-18 by mDC. No significant changes could be observed in surface marker expression or T cell stimulatory capacity, neither in cultures of DC matured under influence of progesterone, estradiol nor betaHCG. CONCLUSIONS: PBMC-derived DC seem to be relatively stable against the influence of pregnancy associated hormones apart from particular effects on cytokine production which partly could contribute to the modification of immune responses observed in normal early pregnancy.