Th17/Treg平衡与正常妊娠及子痫前期关系的研究

目的:探讨Th17及CD4+CD25+Foxp3+调节性T(regulatory T,Treg)细胞介导的免疫反应在子痫前期发病中的作用。方法:选取在山东大学齐鲁医院进行产前检查的子痫前期患者25例及正常晚孕期妇女27例,并选取育龄期健康未孕妇女20例。利用流式细胞技术检测3组患者血样中分泌IL-17的Th17及CD4+CD25+Foxp3+Treg细胞亚群占CD4+T淋巴细胞的百分比,并计算Th17/Treg比率,比较其在子痫前期与正常妊娠时的变化;再用酶联免疫吸附试验(ELISA)检测3组患者外周血中IL-17、IL-10、TGF-β1等细胞因子的表达并进行比较。结果:(1)与正常未孕组及正常妊娠组相比,子痫前期患者外周血中分泌IL-17的Th17细胞占CD4+T淋巴细胞百分比明显升高(P<0.01),CD4+CD25+Foxp3+Treg细胞占CD4+T淋巴细胞百分比明显降低(P<0.01);与正常未孕组相比,正常妊娠组Th17细胞的比例明显降低(P<0.01),Treg细胞比例明显升高(P<0.05);(2)子痫前期患者外周血中Th17/Treg比率明显高于正常未孕组及正常妊娠组(P<0.01),正常妊娠组外周血中Th17/Treg比率与健康未孕组相比明显降低(P<0.01);(3)子痫前期患者外周血中IL-17及TGF-β1的TGF-β1表达明显高于正常未孕组及正常妊娠组(P<0.05),而IL-10的表达未见明显差异(P>0.05);与正常未孕组相比,正常妊娠组外周血中IL-17、IL-10及TGF-β1的水平均未见明显变化(P>0.05)。结论:正常妊娠可能依赖于Treg细胞介导的免疫耐受状态,子痫前期患者外周血中Th17/Treg之间的平衡失调而倾向于Th17介导的促炎状态,这一机制可能参与了子痫前期的发生发展。     

反复自然流产与Th1 Th2细胞因子失衡的研究

目的：探讨Th1/Th2型细胞因子失衡在原因不明反复自然流产（URSA）发生中的作用。方法：采用双抗体夹心酶联免疫吸附实验（ELISA），测定42例原因不明反复自然流产妇女及40例正常经产妇的外周血及蜕膜 中干扰素γ（IEN－γ）和白介素－4（IL－4）的含量。结果：①正常早期妊娠妇女IL－4的诱生水平比正常非妊娠组升高（P＜0．05），反复自然流产妇女IFN－γ的诱生水平比正常非妊娠组升高（P＜0．05），IL－4诱生水平降低（P＜0．05）；②正常早期妊娠妇女的IFN－γ/IL-4的比值比正常非妊娠组下降（P＜0．05），反复自然流产妇女的IFN－γ/IL-4的比值显著升高（P＜0．01），在外周血及蜕膜中的变化是一致的。结论：正常妊娠以Th2型细胞因子反应为主，而在反复自然流产患者中，Th1型细胞因子过度表达，Th1/Th2失去平衡，可能是原因不明反复自然流产发生的原因。     

系统性红斑狼疮患者妊娠期细胞因子的水平及意义

目的:探讨系统性红斑狼疮(SLE)患者妊娠期细胞因子的改变,为临床检测及治疗提供指导。方法:选取单胎妊娠合并SLE患者53例,并选取同期正常单胎妊娠妇女50例作为对照。收集所有对象的妊娠早期(≤12周)及妊娠晚期(≥28周)时血清标本,采用酶联免疫吸附试验检测血清白细胞介素1α(IL-1α)、IL-1β、IL-2、IL-6、IL-8、IL-10、IL-12p70、γ干扰素(IFN-γ)和肿瘤坏死因子α(TNF-α)。结果:对照组妊娠晚期Th1型细胞因子(IL-1α,IL-1β,IL-2,IL-8,IL-12p70,IFN-γ和TNF-α)较妊娠早期明显减少(均P0.05),而Th2型细胞因子(IL-6、IL-10)却没有明显变化(P0.05)。SLE组Th1型细胞因子中只有IL-1α在妊娠晚期明显下降(P=0.001),其他Th1型细胞因子和Th2型细胞因子均无明显改变(P0.05)。在妊娠早、晚期,SLE组的IL-10水平均显著高于对照组,差异有统计学意义(均P=0.000)。对照组妊娠晚期IFN-γ/IL-6比值明显低于妊娠早期,差异有统计学意义(0.65±0.22 vs.1.05±0.17,t=10.17,P=0.000),而SLE组妊娠晚期IFN-γ/IL-6比值与妊娠早期相比差异无统计学意义(0.53±0.16 vs.0.48±0.18,t=1.51,P=0.067)。对照组妊娠早、晚期的IFN-γ/IL-10比值均显著高于SLE组,差异有统计学意义(2.16±0.47 vs.0.79±0.13,t=19.91,P=0.000;3.35±0.96 vs.1.26±0.26,t=14.89,P=0.000)。结论:SLE患者妊娠期细胞因子的变化是复杂多样的,IL-10水平显著高于对照组,妊娠期监测细胞因子对确定SLE疾病活动和判断病情变化有着重要指导意义。     

辅助性T淋巴细胞1和2比率变化与妊娠期高血压疾病发病的关系

目的探讨辅助性T淋巴细胞(Th)1、2比率变化与妊娠期高血压疾病发病的关系。方法采用流式细胞技术,分别检测12例正常未妊娠妇女(正常未孕组)、12例正常妊娠妇女(正常妊娠组)、10例妊娠期高血压患者(高血压组)、25例子痫前期患者(包括10例轻度和15例重度,子痫前期组)的外周血及蜕膜组织(正常未孕组除外)中的Th1/Th2比率。结果正常未孕组妇女外周血中的Th1/Th2比率为10.5±1.5,正常妊娠组妇女外周血及蜕膜组织中的Th1/Th2比率分别为9.5±2.9及7.6±4.6、高血压组妇女分别为12.1±3.4及13.1±5.6、子痫前期组分别为16.8±3.8及26.7±9.4。子痫前期组外周血及蜕膜组织中Th1/Th2比率均明显高于其他各组,分别比较,差异均有统计学意义(P<0.05);且子痫前期组蜕膜组织中Th1/Th2比率明显高于外周血,两者比较,差异有统计学意义(P<0.001)。高血压组妇女的Th1/Th2比率变化处于子痫前期组和正常妊娠组之间。结论妊娠期高血压疾病患者Th1/Th2比率升高,导致Th1/Th2比率平衡紊乱可能是妊娠期高血压疾病发生的重要原因。     

干预CD86协同刺激信号对母胎界面Th1/Th2型细胞因子转录调控及妊娠结局的影响

目的探讨干预CD86协同刺激信号对母胎界面Th1/Th2型细胞因子转录调控及妊娠结局的影响。方法:将正常妊娠模型(CBA/J×BALB/c)和自然流产模型(CBA/J×DBA/2J)CBA孕鼠均分为两组:对照组(各10只)于孕d 4、d 6、d 8腹腔注射大鼠IgG;干预组(各10只)于孕d 4、d 6、d 8腹腔注射大鼠抗小鼠CD86 mAb。孕d 9竞争性半定量RT-PCR测定各组母胎界面组织中Th1型(IL-12、IFN-γ)/Th2型(IL-4、IL-10)细胞因子转录水平;孕d 12比较两种模型各组的胚胎吸收率。结果:正常妊娠模型中,干预CD86协同刺激信号对母胎界面Th1/Th2型细胞因子转录水平及妊娠预后均无显著影响(P>0.05)。自然流产模型中,干预CD86协同刺激信号能够升调节母胎界面局部Th2型而降调节Th1型细胞因子转录水平,并显著改善其妊娠预后(P<0.05)。结论:于孕早期干预CD86协同刺激信号能够调控母胎界面局部Th1/Th2型细胞因子转录,形成维持正常妊娠所需的Th2型免疫偏倚,诱导母胎免疫耐受。     

Th1/Th2类细胞因子与葡萄胎恶变的相关性研究

目的:通过对妊娠滋养细胞疾病患者外周血血清中Th1/Th2类细胞因子的测定,从免疫学角度探讨Th1/Th2类细胞因子预测葡萄胎恶变的价值。方法:收集1997至2001年首次以葡萄胎诊断进行清宫和手术的患者的外周血血清,采用酶联免疫吸附法检测血清中IFN-γ、IL-2、IL-4、IL-10的水平来确定Th1/Th2的活性。结果:与侵葡+绒癌组相比,葡萄胎组IFN-γ含量明显增加(P<0.001),IL-10含量明显降低(P<0.001)。葡萄胎患者中,与非恶变组相比,恶变组Th1类细胞因子(IFN-γ,IL-2)含量呈下降趋势,而Th2类细胞因子(IL-4,IL-10)含量呈增加趋势,4种细胞因子变化均具有统计学差异(P<0.0001)。随着滋养细胞增生程度的增加,4种细胞因子的变化均具有统计学差异,除外IL-4的变化在轻、中度增生组中无统计学意义(P>0.05)。结论:Th1/Th2细胞因子向Th2漂移时,葡萄胎恶变倾向增高。     

阿奇霉素治疗沙眼衣原体感染孕鼠对其妊娠结局及免疫状态的影响

目的:了解沙眼衣原体感染孕鼠的妊娠结局与体内Th1型、Th2型细胞因子水平的关系,以及阿奇霉素干预治疗对其免疫状态及妊娠结局的影响,探讨沙眼衣原体致不良妊娠结局的免疫机理。方法:随机将孕鼠分成3组,于孕2、3、4天,对照组阴道内接种无感染衣原体由McCoy细胞制备的SPG悬液30μl,模型组阴道内接种1.0×106IFU/ml沙眼衣原体F型株的SPG悬液30μl,给药组阴道接种衣原体后于妊娠第8天腹腔注射阿齐霉素10mg/kg(单次给药)。孕12天用酶联免疫法检测各组小鼠血清中Th1型(IFN-γ、TNF-α)/Th2型(IL-4、IL-10)细胞因子表达水平,计算IL-4/TNF-α,IL-10/TNF-α比率。用方差分析和χ2检验比较3组的妊娠失败率、胚胎吸收率、胎重、孕期体重增加量等,以及3组之间细胞因子水平及Th1/Th2型免疫调节平衡的比率等。结果:模型组与对照组比较体内Th2型细胞因子下降(IL-10),而Th1型细胞因子上升(TNF-α),且代表Th1/Th2型免疫调节平衡的细胞因子比值:IL-4/TNF-α,IL-10/TNF-α模型组显著低于对照组,形成Th1型免疫偏倚。给药组降低了妊娠失败率并降低了Th1型免疫偏倚程度。模型组小鼠妊娠失败率及反复妊娠失败率显著高于对照组;给药组小鼠妊娠失败率较模型组低而较对照组高,反复妊娠失败率较模型组下降明显(P0.05);3组的胚胎吸收率,胚重,孕期体重增加量无显著差异。结论:阴道接种沙眼衣原体使体内细胞因子向Th1型漂移,增加了小鼠的妊娠失败率;阿奇霉素干预可降低Th1型漂移程度,降低小鼠妊娠失败率。     

子痫前期患者辅助性T淋巴细胞及协同刺激分子CD28/CTLA-4的表达

目的:探讨辅助性T淋巴细胞及协同刺激分子CD28/CTLA-4在子痫前期发病中的作用。方法:采集22例正常妊娠妇女(正常妊娠组)及45例子痫前期患者(子痫前期组,包括22例轻度和23例重度)外周血,用流式细胞技术分别检测外周血Th1、Th2及Th1/Th2比率及CD28和CTLA-4在CD4+T细胞的表达。结果:子痫前期组Th1、Th1/Th2、CTLA-4均高于正常妊娠组,且重度高于轻度(P0.05);子痫前期组Th2、CD28/CTLA-4低于正常妊娠组,且重度低于轻度(P0.05)。CD28与Th1呈负相关,r=-0.295;CTLA-4与Th1呈正相关,r=0.551,与Th2呈负相关,r=-0.363;CD28/CTLA-4与Th1/Th2呈负相关,r=-0.707。结论:子痫前期患者外周血协同刺激分子CD28/CTLA-4表达异常,高表达的CTLA-4促进CD4+T淋巴细胞过度活化并向Th1亚群分化,导致Th1/Th2比率失衡,这可能是子痫前期发生的重要原因之一。     

泰山磐石散对复发性流产小鼠母胎界面Th1/Th2细胞因子及妊娠预后的影响

目的:观察泰山磐石散对复发性流产小鼠母胎界面Th1/Th2细胞因子及妊娠预后的影响,为泰山磐石散临床应用提供新的实验依据。方法:采用经典造模方式DBA/2小鼠与CBA/J杂交,获得复发性流产小鼠模型,随机将与DBA/2小鼠合笼的60只CBA/J妊娠小鼠分为模型组、中药低剂量组、中药中剂量组、中药高剂量组和阳性对照组;与BALB/C合笼的10只CBA/J孕鼠作为正常妊娠模型。于妊娠14 d后处死孕鼠,观察小鼠胎盘丢失情况,并提取培养胎界母细胞,24 h后收集细胞上清液,酶联免疫吸附测定(ELISA)法检测上清液中肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)、白细胞介素4(IL-4)和IL-10含量。结果:经泰山磐石散治疗后,与模型组比较,复发性流产小鼠胎盘丢失率明显改善,母胎界面细胞上清液中Th1型细胞因子IFN-γ明显降低,Th2型细胞因子IL-4、IL-10明显升高,Th1/Th2免疫调节失衡明显改善,以中药高剂量组改善最为明显。结论:泰山磐石散能改善复发性流产小鼠胎盘丢失情况,其具体机制可能是通过调节Th1/Th2免疫调节平衡实现。     

先兆子痫患者外周血Th1 Th2细胞亚群功能变化研究

目的：探讨先兆子痫患者Th1、Th2细胞亚群功能变化。方法：采用ELISA方法检测17例先兆子痫患者及15例正常孕妇PBMC培养上清液中IFN－γ和IL－4的水平。结果：先兆子痫患者IFN－γ水平升高，差异非常显著（P＜0．01）；IL－4水平下降，差异非常显著（P＜0．001）；IFN－γ／IL－4比值升高，差异非常显著（P＜0．001）；且IFN－γ水平与平均动脉压呈显著正相关，γ为0．546（P＜0．01）；IL－4水平与平均动脉压呈显著负相关，γ为－0．544（P＜0．01）；IFN－γ/IL-4比值与平均动脉压呈显著正相关，γ为0．786（P＜0．01）。结论：先兆子痫患者Th1、Th2细胞亚群功能失衡，与病情密切相关。     

Th1, Th2, Th17 and Treg levels in umbilical cord blood in preeclampsia

Introduction: Preeclampsia is one of the major causes of maternal and neonatal mortality. During pregnancy, the immune system must maintain the tolerance to the fetus, thus changes in the cytokine balance may result in a disturbed pregnancy. T helper cells play an important role in modulation of the immune system and are involved in this cytokine balance.

Objective: Many studies have been performed to study the T cell composition in different compartments during pregnancy, although this is the first study in which T cells are evaluated in umbilical cord blood.

Study design: Intracellular expression of INF-gamma, IL-17, IL-4 and forkhead foxP3 in CD4+ T cells was evaluated in umbilical blood from healthy pregnant and preeclamptic women using a flow cytometer.

Results: Th2 and Treg cells levels were significantly diminished in preeclamptic compared to the healthy women, but no difference in Th1 and Th17 levels were found between both groups.

Conclusions: Our data suggest that the cytokine balance is broken, encouraging the development of an exacerbated inflammatory response. Our results show that there is a shift, in the Th1/Th2, and the Th17/Treg balance, favoring skewness towards a proinflammatory status in the umbilical cord blood in preeclampsia.     

The predominance of Th17 lymphocytes and decreased number and function of Treg cells in preeclampsia

The aim of this study was to estimate the prevalence of CD3(+)CD4(+) T lymphocytes producing IL-17, IL-2, IFN-γ, and IL-4, plus CD4(+)CD25(+)FoxP3(+) T regulatory (Treg) cells, in peripheral blood of patients with preeclampsia and healthy women in the third trimester of normal pregnancy. Another purpose was to assess the immunosuppressive activity of Treg cells from patients with preeclampsia compared with controls. Thirty-four preeclampsia patients and 27 healthy pregnant women were included. The percentages of CD4(+)CD25(+)FoxP3(+) Treg cells and CD3(+)CD4(+) T lymphocytes with intracellular expressions of cytokines were estimated using monoclonal antibodies and flow cytometry. In vitro functional assays were performed using a Treg Cell Isolation Kit and (3)H-thymidine incorporation assays. The percentage of CD3(+)CD4(+) T lymphocytes producing IL-17A was significantly higher in preeclampsia than in healthy, normotensive pregnant women in the third trimester (p<0.001). The population of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower in the study group compared with controls (p<0.05). There was no change in the stimulation index of CD3(+)CD4(+)CD25(-) T lymphocytes from preeclampsia patients without Treg cells and after addition of autologous Treg cells. In normal pregnancy, the stimulation index of CD3(+)CD4(+)CD25(-) T lymphocytes was significantly higher without Treg cells compared with the response after addition of autologous Treg cells (p<0.05). The results suggest up-regulation of the Th17 immune response in preeclampsia. The decreased number and function of Treg cells may be responsible for activating the inflammatory response characteristic of this disorder. In preeclampsia, the predominance of Th17 immunity could act through modulating the Th1/Th2 immune balance.     

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.     

Lymphocyte subpopulations in gestosis

Lymphocyte subsets from peripheral venous blood of 11 patients with preeclampsia were enumerated by monoclonal antibodies and compared to the values of 10 normal term pregnant women. All women were in their last trimester of pregnancy. The number of B and T cells was similar in both groups. In the preeclampsia group, more helper T cells could be shown, whereas the number of suppressor T cells was reduced. Thus, the helper/suppressor ratio was increased from 2.01 in normal pregnancies to 3.97 in preeclamptic patients. In two patients with a true-positive angiotensin stress test similar changes as in preeclampsia could be seen. In only one out of five patients with preeclampsia the helper/suppressor ratio returned to normal values four to five weeks after delivery.     

Cytokine Production by Monocytes,NK Cells,and Lymphocytes Is Different in Preeclamptic Patients as Compared with Normal Pregnant Women

Objective. To measure cytokine production in ex vivo stimulated leukocyte populations of women with normal pregnancy and those with preeclampsia. Methods. Whole blood from preeclamptic and normal pregnant women was stimulated with LPS or PMA/Ca-ionophore. The percentages of IFNγ and IL-2, 4, and 10 producing lymphocytes and NK cells and the percentages of TNFα, IL-1β, and IL-12 producing monocytes were measured by flowcytometry. Results. In women with preeclampsia, there was a significantly increased percentage IL-4 producing cytotoxic T cells. Also, a significant decreased percentage IL-2 producing T helper cells and IL-12 producing monocytes was seen as compared with normal pregnancy. Conclusion. Th1 cytokine production of lymphocytes and monocytes appears to be decreased in our group of preeclamptic patients compared with normal pregnant women.     

Dendritic cells in the circulation of women with preeclampsia demonstrate a pro-inflammatory bias secondary to dysregulation of TLR receptors

Toll-like receptors (TLRs) are central components of the innate immune system that recognize both microbial ligands and host products released during tissue damage. Data from epidemiologic studies and animal models suggest that inappropriate activation of the immune system plays a critical role in the development of preeclampsia. This study evaluates in a systematic fashion the expression and function of TLRs in the circulation of patients with preeclampsia compared to healthy pregnant controls. We evaluated TLR expression and function in primary dendritic cells (DCs) of 30 patients with preeclampsia and 30 gestational age-matched healthy pregnant controls. DCs were stimulated with the different TLR ligands engaging TLR1/2, TLR2/6, TLR3, TLR4, TLR5, TLR7, TLR8 and TLR9. The expression of TLR-induced production of TNF-α, IFN-α, IL-6, and IL-12 were measured by multicolor flow cytometry. Basal expression of TLR3, TLR4 and TLR9 was significantly increased in DCs isolated from women with preeclampsia. Preeclamptic DCs also expressed significantly higher basal levels of cytokines. In contrast, preeclamptic DCs demonstrated a less robust response to stimulation with various TLR ligands as compared with healthy pregnant controls. Under basal conditions, DCs from preeclamptic individuals express higher levels of select TLRs and produce more pro-inflammatory cytokines as compared with healthy controls. As such, the ability of these cells to mount an inflammatory reaction in response to a TLR ligand is limited. These data demonstrate a dysregulated pattern of TLR expression and cytokine production in DCs from PE patients that may limit further activation by TLR engagement.     

Expression and significance of dendritic cells and Th17/Treg in serum and placental tissues of patients with intrahepatic cholestasis of pregnancy

Purpose: Dendritic cells (DCs) are involved in immune system, which can also regulate the differentiation of T helper 17 (Th17) and regulatory T cells (Treg). DCs and Th17/Treg participate in preeclampsia and recurrent spontaneous abortion (RSA), but there is still lack of research in intrahepatic cholestasis of pregnancy (ICP). The aim was to evaluate the expression and significance of CD83⁺DCs, CD1a⁺DCs, interleukin-17 (IL-17) and IL-35 in serum and placental tissues of patients with ICP.

Methods: Thirty cases of mild ICP, 25 cases of severe ICP were selected, and 30 cases of normal pregnant women were selected as control group. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) were used to detect the expression of CD83⁺DCs, CD1a⁺DCs, IL-17 and IL-35 in serum and placenta tissues, respectively.

Results: There were more CD83⁺DCs, IL-17 expressed in placenta from women with ICP than in normal pregnancies, while the number of decidual CD1a⁺DCs, IL-35 was significantly lower in ICP than in normal pregnant women. The comparison within three groups had statistical difference (p?+DCs and CD1a⁺DCs levels had no significance. IL-17 was higher in ICP, while IL-35 was lower.

Conclusions: DCs are involved in damaging the maternal–fetal immune tolerance by changing the phenotype and mature state, which may affect the differentiation of Th17/Treg to cause ICP.     

Blood myeloid and lymphoid dendritic cells are stable during the menstrual cycle but deficient during mid-gestation

The aim of this study was to estimate the populations of peripheral blood myeloid and lymphoid dendritic cells (CD1c(+), BDCA-2(+), BDCA-4(+)) and the CD1c(+):BDCA-2(+) ratio in phases of the ovarian cycle and in normal pregnant patients. 18 non-pregnant women and 17 normal pregnant women were included. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies (mAbs) against blood dendritic cell antigens (anti-BDCA-1, BDCA-2, BDCA-4) and estimated using flow cytometry. CD1c(+), BDCA-2(+) and BDCA-4(+) dendritic cells were present in the follicular and luteal phases of the ovarian cycle and in all trimesters of normal pregnancy. The percentages of CD1c(+) dendritic cells did not differ between the follicular and luteal phases of the ovarian cycle. The percentage of BDCA-2(+) dendritic cells was lower in the luteal phase of the ovarian cycle compared with the follicular phase, but the differences were not statistically significant. The CD1c(+):BDCA-2(+) cell ratio was significantly lower in the luteal phase compared with the follicular phase of the ovarian cycle. The numbers of dendritic cells were significantly lower in the second trimester when compared with the first and third trimesters of normal pregnancy. Furthermore, in the second trimester, the CD1c(+):BDCA-2(+) ratio was higher than in the other trimesters of normal pregnancy. All populations of dendritic cells and the CD1c(+):BDCA-2(+) ratio did not differ in the first and third trimesters of physiological pregnancy. Our results suggest that myeloid and lymphoid dendritic cells are not affected by steroid hormones during the menstrual cycle. The deficiency of peripheral blood dendritic cells observed during the second trimester of normal pregnancy can be associated with their migration to the uterus during the second physiological invasion by cytotrophoblast.     

Lymphocyte subset distribution and cytokine secretion in third trimester decidua in normal pregnancy and preeclampsia

BACKGROUND: Excessive Th1 activity in peripheral blood plays a probable role in the pathogenesis of preeclampsia. The aim of the study was to investigate whether disturbed local immune reactions are also present in decidua. METHODS: Flow cytometric analysis of CD3, CD19, CD56/CD16, CD4, CD8, CD4/CD29, CD4/CD45RA, CD4/CD45RO, CD8/CD28, CD3/CD69 lymphocyte subsets isolated from third trimester decidua of pregnants with preeclampsia (n=21) and pregnant controls (n=11) subjected to elective caesarean sections. Spontaneous and phytohemaglutynine stimulated "in vitro" secretion of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta by decidual lymphocytes was studied by ELISA. For the statistical significance of differences between the groups the U Mann-Whitney test was performed (confidence interval P<0.05). RESULTS: Preeclamptic patients were characterized with an increased percentage of the CD3-/CD56+CD16+, CD8+/CD28+ and decreased percentage of CD3+, CD19+, CD4+/CD45RA+ lymphocytes. The profile of secreted cytokines shifts in favor of Th1 activity (extremely high IFN-gamma and low IL-6 and IL-10 secretion). Decidual IL-12 secretion in preeclamptic patients is decreased compared to controls. CONCLUSION: Changes in NK and T lymphocyte subsets followed with Th1 cytokine IFN-gamma over-activity, could affect local immunoregulatory mechanisms in third trimester decidua of preeclamptic patients.     

正常月经周期妇女和自然流产妇女外周血中树突状细胞亚群的变化

目的:探讨正常月经周期妇女和自然流产妇女外周血中髓样树突状细胞(myeloid dendritic cell,MDC)和浆细胞样树突状细胞(plasmacytoid dendritic cell,PDC)的变化。方法:选择正常月经周期妇女30例,分别在每个妇女月经周期卵泡期和黄体期采集外周血;早期自然流产妇女30例流产后清宫前采集外周血,并以正常早期妊娠妇女为对照组。应用流式细胞术,检测各组外周血单个核细胞中MDC和PDC的百分率及MDC/PDC比率。应用放射免疫法检测各组外周血中雌二醇(E_2)和孕酮(P_4)的水平。结果:正常月经周期黄体期外周血中MDC的百分率和MDC/PDC比率显著低于卵泡期(P<0.01),PDC的百分率黄体期与卵泡期无统计学差异(P>0.05)。正常月经周期妇女黄体期外周血中E_2和P_4水平显著高于卵泡期(P<0.01)。早期自然流产妇女外周血中MDC的百分率和MDC/PDC比率显著高于正常早孕组(P<0.01),而PDC的百分率与正常早孕组无统计学差异(P>0.05)。自然流产组E_2和P_4水平显著低于正常早孕组(P<0.01)。结论:早期自然流产妇女外周血中MDC的百分率和MDC/PDC比率显著升高,可能参与了导致母体对胎儿发生免疫排斥。