Within-woman change in regulatory T cells from pregnancy to the postpartum period

Regulatory T cells (Treg cells) are an important area of investigation in human health and disease. In this study, the trajectory of percentage of Treg cells (defined as CD4+CD25+Foxp3+CD127--lymphocytes) was measured in the blood of 208 women during pregnancy and up to three additional times in the postpartum period (1, 6 and 12 months postpartum). Whether the trajectory was affected by gravidity, parity, neonatal sex, pet exposure, maternal atopic and asthma status, smoking, maternal race or other pregnancy factors was examined. Multilevel models were fit using full maximum likelihood methods and included both random and fixed effects. Overall, percentages of Treg cells increased from the prenatal to the postpartum period. Among women who were not atopic, nulliparous women had lower percentages of Treg cells over time compared with parous women. Atopic women with pets in the home during pregnancy had lower percentages of Treg cells than atopic women who did not have pets. The trajectory was not affected by the other factors investigated. We conclude that within-woman change in percentages of Treg cells may vary by time in relation to delivery, as well as by maternal atopic status and exposure to pets and number of prior births. The data did not indicate an overall decline in Treg cells in the postpartum period. Future work to better identify the role of Treg cells in successful pregnancy would ideally include a set of well characterized women sampled serially starting prior to pregnancy and throughout the postpartum period.     

Urinary pregnandiol-3-glucuronide and estrone conjugates to creatinine ratios in early pregnancies complicated by vaginal bleeding

There is no simple and rapid test available to predict the outcome of an early pregnancy complicated by vaginal bleeding. In this prospective study, 15 women with normal pregnancies collected a weekly urine sample between 6 and 13 weeks' gestation. A single random urine sample was obtained from 15 women with bleeding who continued to carry their child and 50 women who proceeded to have a spontaneous abortion (SAB). Pregnandiol-3-glucuronide (PDG) was determined with the use of enzyme-multiplied immunoassay technique (EMIT) and estrone conjugates (E1C) were measured by radioimmunoassay (RIA). The ratios of these metabolites to creatinine (C) were calculated. PDG/C ratios in normal women rose gradually from 6 weeks on. All women with bleeding during a normal pregnancy had ratios in the normal range, but 94% of women with a SAB had ratios below the normal range. The E1C/C ratio remained unchanged from 6 to 11 weeks and then rose rapidly. Until 11 weeks, there was no clear separation between the E1C/C ratios of the women with a SAB and the women with bleeding who continued their pregnancies. The prognosis of threatened abortion can be made by a urinary PDG/C ratio but not by an E1C/C ratio. EMIT is simple and quick and uses technology present in many laboratories.     

Pregnancy-associated changes in peripheral blood lymphocyte subpopulations in normal Kuwaiti women.

It has recently been reported that healthy pregnancy is associated with systemic immunosuppression. The aim of this study was to evaluate the numbers and distribution of lymphocyte subpopulations in normal, healthy pregnant Kuwaiti women. Thirty-four healthy normotensive women in the 3rd trimester of pregnancy were studied using flow cytometry to define lymphocyte subpopulations and were compared with 16 non-pregnant women. A decrease in the absolute numbers of lymphocytes was observed affecting T cells (CD3+, CD4+, CD8+), B cells (CD19+), and natural killer cells (CD16+/CD56+). When analyzed as a percentage of the total lymphocyte population, there was a significant decrease in B cells and an increase in CD4+ T cells. The T cell population revealed increased expression of CD25 on CD4+ and CD8+ cells, of HLA-DR on CD8+ cells, and of CD54 on CD4+ T cells. The reduced number of lymphocytes suggests that Kuwaiti females may be immunosuppressed in the 3rd trimester of pregnancy. The presence of activated CD4+ T cells could indicate the expression of a regulatory suppressor T cell population, as Treg cells are CD4+CD25+, and suppressor T cells are thought to be CD8+. Future work is required to explore the significance of these T cell populations in pregnancy.     

CD4~+CD25~+FOXP3~+调节性T淋巴细胞数量及功能变化与子宫内膜异位症患者发病相关性研究

目的:分析子宫内膜异位症(EMs)患者在位、异位内膜及外周血内CD4~+CD25~+FOXP3~+调节性T淋巴细胞(Treg细胞)的数量、分布及功能特性,探讨Treg细胞与EMs发病之间的关系。方法:收集EMs患者在位、异位内膜组织及外周血标本,并以非内异症患者作为对照,免疫组化分析在位、异位内膜内叉头状/翅膀状螺旋转录因子3(FOXP3)阳性细胞的数量及分布变化情况,并与EMs患者的r AFS临床分期进行相关性分析。流式细胞术检测Treg细胞占外周血CD4~+T淋巴细胞的百分比,及磁珠分选外周血内CD4~+CD25~+T细胞及CD4~+CD25-T细胞,3H-thymidine法测定CD4~+CD25~+T细胞免疫抑制功能变化情况。结果:EMs、非EMs患者在位内膜内平均FOXP3阳性细胞密度并无显著差异[(76.44±62.14)/mm~2vs(50.59±20.79)/mm~2;WU=152.0,P=0.20]。进一步按月经周期行亚组分析:EMs分泌期内膜内FOXP3阳性细胞密度显著高于非EMs患者[(94.84±53.91)/mm~2vs(31.37±19.02)/mm~2;MU=43.00,P=0.03]。不同r AFS期别患者卵巢异位症病灶内FOXP3阳性细胞密度并无显著差异[I~II期:(123.00±115.00)/mm~2vs III~IV期:(111.00±108.00)/mm~2;MU=139.5,P0.05]。EMs患者外周血内Treg细胞比例显著低于非EMs患者[(0.58±0.21)%vs(1.35±0.38)%,P0.001],但EMs患者外周血CD4~+CD25~+T淋巴细胞对CD4~+CD25-T淋巴细胞增殖抑制率,与非EMs相比并无显著变化[(68.43±18.15)%vs(66.37±17.78)%,P0.05]。结论:EMs患者在位内膜内Treg细胞失去正常的周期波动性,EMs患者分泌期内Treg细胞数目增加可能与内异症发病相关。EMs患者外周血内Treg细胞比例下降,但其免疫抑制功能并无明显改变。     

哮喘患儿诱导痰辅助性T细胞亚群及相关细胞因子变化研究

探讨支气管哮喘患儿诱导痰中辅助性T细胞（Th）亚群及相关细胞因子的变化及意义。方法　观察2009年9月至2010年1月深圳儿童医院收集的20例哮喘患儿及20例同年龄对照组儿童,采用流式细胞术检测哮喘患儿外周血Th1、Th2、Th17、CD4+CD25+调节性T细胞（Treg）数量;同时收集哮喘患儿和对照组儿童的诱导痰,进行炎性细胞计数与分类,荧光定量聚合酶链反应（Real Time PCR）测定外周血单个核细胞及诱导痰T-bet、IFN-γ、GATA3、IL-4、Foxp3、TGF-β、RORrt、IL-17A、IL-8、TNF-α等mRNA表达水平。结果　（1）急性期哮喘患儿外周血Th1、Treg细胞比例降低（P < 0.05）、Th2细胞、Th17细胞比例与正常对照组差异无统计学意义（P > 0.05）。（2）急性发作期哮喘患儿外周血单个核细胞T-bet、Foxp3、IFN-γ、TGF-β表达降低（P < 0.05）,GATA3、RORγt、IL-4、IL-17A 与正常对照组差异无统计学意义（P > 0.05）。（3）急性发作期哮喘患儿诱导痰中性粒细胞、嗜酸性粒细胞数量增加（P < 0.05）。（4）急性发作期哮喘患儿诱导痰中RORγt、IL-17A、GATA3、IL-4、IL-5、IL-8、TNF-α表达增高（P < 0.05）,T-bet、IFN-γ、Foxp3、TGF-β表达与正常对照组差异无统计学意义（P > 0.05）。结论　哮喘患儿诱导痰Th2与Th17转录因子和细胞因子增多,可能是导致儿童哮喘气道炎症反应的重要原因。     

Effect of menstrual cycle variation in female sex hormones on cellular immunity and regulation

Ovarian steroid hormones reduce cell-mediated immunity (CMI), perhaps by increasing regulatory T cells. We examined the relationship of estrogen and progesterone plasma concentrations during the menstrual cycle with circulating regulatory T cells (Treg cells) and with varicella-zoster virus (VZV)-specific lymphocyte proliferation (VZV-LPA). Twenty healthy and 20 HIV-infected women were tested at 1-4, 10-14, and 20-24days of the menstrual cycle. HIV-infected women experienced significant increases in the frequency of peripheral blood CD4+IL10+ and CD8+FoxP3+ Treg cells from the early and late follicular phases to the luteal phase of their cycles. Healthy women experienced significant increases only in CD4+IL10+ Treg cells. The increase in CD4+IL10+ Treg cells between the late follicular and the luteal phases of HIV-infected and uninfected women significantly correlated with the corresponding increases in progesterone plasma concentrations. VZV-LPA results decreased from the early and late follicular phases to the luteal phase in both groups. The decrease in VZV-LPA results significantly correlated with the increase in CD4+IL10+ Treg cells underscoring the potential immunosuppressive effect of the progesterone-stimulated Treg cells. In conclusion, the increase in progesterone levels during the menstrual cycle was associated with higher Treg frequencies and lower CMI.     

TGF-β1在早孕妇女外周血和蜕膜组织中诱导生成T调节性(Treg)细胞

目的:研究TGF-β1是否能在人母-胎界面诱导生成T调节性(Treg)细胞。方法:早孕妇女外周血和蜕膜CD4+CD25-T细胞中加入不同浓度的TGF-β1(0 ng/ml、2 ng/ml、5 ng/ml、10 ng/ml)分别于培养后第2日、第4日和第6日用流式细胞仪检测培养Foxp3的表达情况,随后将诱导生成的CD4+Foxp3+T细胞与CD8+T细胞混合培养,观察后者凋亡因子CD95配体(CD95L)的表达情况。结果:体外培养中,TGF-β1可诱导早孕妇女的外周血和蜕膜CD4+CD25-T细胞生成诱导性Treg细胞,随着培养时间增加而增强诱导效应,且在TGF-β1浓度为5 ng/ml时诱导功能最强;诱导生成的CD4+Foxp3+T细胞具有促进效应细胞CD8+T细胞凋亡的功能。结论:体外培养中,TGF-β1能将人外周血和蜕膜诱导生成Treg细胞,且具有免疫抑制功能,有良好的免疫治疗前景。     

卵巢早衰患者外周血CD4~+CD25~+Treg细胞的变化及意义

目的:探讨CD4+CD25+调节性T细胞(即CD4+CD25+Treg细胞)在卵巢早衰发病机制中的作用。方法:流式细胞仪定量检测卵巢早衰(premature ovarian failure,POF)患者、卵巢储备功能下降(diminished ovarian reserve,DOR)患者及健康对照组外周血CD4+T、CD8+T细胞及CD4+CD25+Treg细胞数量,应用3H-thymidine掺入法测定POF患者及对照组外周血CD4+CD25+Treg细胞对效应性T细胞的增殖抑制功能。结果:与对照组相比,POF患者及DOR患者CD4+CD25+Treg细胞比例降低(P<0.01)、POF患者CD4+T/CD8+T细胞比值增高(P<0.05),DOR患者CD4+T/CD8+T细胞比值无明显变化(P>0.05);POF患者免疫抑制功能无明显降低(P>0.05)。结论:CD4+CD25+Treg细胞比例降低与T细胞亚群失衡可能是POF的发病机制。     

Decreased frequency of peripheral blood CD8+CD25+FoxP3+regulatory T cells correlates with IL-33 levels in pre-eclampsia

Objective: We aimed to investigate the role of CD8⁺CD25⁺Foxp3⁺regulatory T (Treg) cells in pre-eclampsia (PE).

Methods: This was a cross-sectional study of 46 patients with PE and 24 normotensive women within the third trimester of gestation. We analyzed the percentages of CD8⁺CD25⁺Foxp3⁺Treg cells in peripheral blood using flow cytometry and the serum levels of interleukin (IL)-6, IL-17A, IL-10, TGF-β1, IL-1β, and IL-33 by Luminex 200.

Results: We found that patients with PE had lower percentages of CD8⁺CD25⁺Foxp3⁺Treg cells than normotensive pregnant women. In addition, the percentage of CD8⁺CD25⁺Foxp3⁺Treg cells was positively correlated with IL-33 concentration and negatively correlated with IL-17A concentration in patients with PE. We also found that IL-33 treatment can induce proliferation of CD8⁺CD25⁺Foxp3⁺Treg cells in vitro.

Conclusions: These findings suggest that the reduced CD8⁺CD25⁺Foxp3⁺Treg cells may play a role in the pathogenesis of PE.

Abbreviations

PE: pre-eclampsia; PBMCs: peripheral blood mononuclear cells; CTLA-4: cytotoxic T-lymphocyteantigen-4; APCs: antigen presenting cells; TGF-β: transforming growth factor-β; IL: interleukin; Treg: cells regulatory T cells; PBS: phosphate-buffered saline; Foxp3: forkhead Box protein 3; HELLPs: hemolysis, elevated liver enzyme and low platelet syndrome     

体外受精-胚胎移植反复失败患者外周血中CD4~+CD25~+Treg的表达

目的:探讨外周血中CD4+CD25+调节性T淋巴细胞(CD4+CD25+Treg)在体外受精-胚胎移植反复失败(RIF)发病机制中的作用。方法:用流式细胞分析技术检测12例RIF组、15例胚胎移植妊娠组、15例正常未孕组患者外周血中CD4+CD25+Treg的表达。结果:RIF组患者外周血中CD4+CD25+Treg的表达率为(0.80±0.56)%,低于正常未孕组的(4.05±0.91)%,两组差异有统计学意义(P<0.05);胚胎移植妊娠组患者外周血中CD4+CD25+Treg的表达率为(11.01±2.09)%,高于RIF组,两组差异有统计学意义(P<0.05)。结论:辅助生殖中反复植入失败可能与CD4+CD25+Treg的表达下降有关,这为免疫治疗提供了理论依据。     

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。     

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.     

CD4~+T细胞亚群表达失衡在妊娠期哮喘发病中的作用及意义

目的:观察妊娠期哮喘小鼠模型中Th17及Treg细胞活性分布和表达变化,探讨Th17/Treg细胞免疫平衡在妊娠期哮喘小鼠发病中的作用机制。方法:24只6～8周龄,雌性BALB/c小鼠随机分为4组,每组6只。以OVA/Al(OH)3混悬液致敏与激发构建妊娠期哮喘组(AP组)和非妊娠期哮喘组(ANP组)小鼠模型;健康妊娠组(HP组)和健康非妊娠组(HNP组)小鼠致敏与激发均以只含Al(OH)3的生理盐水混悬液替代。光镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞计数及分类;HE染色评价小鼠气道炎症程度及肺组织病理学变化;流式细胞术测定小鼠外周血Th17及Treg细胞所占CD4+T细胞比例,计算Th17/Treg细胞比值;RT-PCR检测小鼠肺组织中Th17及Treg细胞相关因子表达。结果:妊娠期哮喘小鼠BALF中炎症细胞分类及计数、肺组织病理学表现和活体肺功能气道高反应性Penh值等客观指标证实,妊娠哮喘小鼠模型构建成功。外周血中,AP组CD4+IL-17+T细胞(Th17)显著升高(P<0.01);Foxp3+T细胞(Treg)显著降低(P<0.01);与HP,HNP组相比,存在Th17/Treg比例失衡(P<0.01)。AP组小鼠肺组织中IL-17、IL-23,IL-10 mRNA表达水平均显著高于HP组及HNP组(P<0.01);IFN-γmRNA则显著低于HP组及HNP组(P<0.01);ANP组各因子mRNA水平变化与AP组一致,组间无显著差异。结论:妊娠期哮喘小鼠存在细胞免疫功能失调;Th17、Treg细胞数量及免疫平衡状态发生改变,可能是妊娠期哮喘发病过程中一个重要的决定因素。     

CD4(+)CD25high regulatory T cells in human pregnancy

In both rodent and human systems, there is an emerging consensus that immunoregulatory activity specific for donor alloantigens is enriched in the CD4(+)CD25+ T cell population. The absence of CD4(+)CD25+ regulatory T (Treg) cells induces severe immunodeficiency with autoimmune disease, dermatitis and fatal infections in humans and mice. CD4(+)CD25+ Treg cells play a critical role in peripheral tolerance, transplantation tolerance and maternal tolerance to the fetus. Although both human and mouse CD4(+)CD25+ Treg have potent regulatory properties, surface phenotypes of human CD4(+)CD25+ Treg cells are not exactly the same as those of mouse CD4(+)CD25+ Treg cells. Murine CD4(+)CD25+ T cells are homogenous and exhibit regulatory function. On the other hand, CD4(+)CD25high T cells are the only cells which exhibit regulatory function in humans. Humans CD4(+)CD25low cells have no ability for immunosuppression. CD4(+)CD25high T cells inhibit the immunostimulation of conventional T cells through cell-to-cell contact or immunosuppressive cytokines such as interleukin 10 and transforming growth factor-beta. As another mechanism of immunosuppression, CTLA-4 on CD4(+)CD25+ regulatory T cells up-regulate indoleamine 2,3-dioxygenase (IDO) expression in dendritic cells which play important roles for immunosuppression. Here, we review the differences between humans and mouse Treg cells and the role of CD4(+)CD25+Treg during pregnancy.     

复发性流产患者外周血T淋巴细胞亚群及CD4^＋辅助性T淋巴细胞亚型的变化

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The immunological profile of infertile women after repeated IVF failure (preliminary study)

OBJECTIVE: The aim of this preliminary study was to estimate the immunological profile of patients after repeated IVF failures. MATERIALS AND METHODS: Seventeen women after repeated IVF failure and 10 non-pregnant women with a history of successful IVF pregnancies were included in the study. We estimated the presence of the auto-antibodies, such as: antinuclear antibodies (ANA), antithyroid antibodies (ATA), antiphospolipid antibodies (APA), antismooth muscle antibodies (ASMA), and antisperm antibodies (ASA). Furthermore, we estimated the percentage of B-1 CD 19+5+ lymphocytes and NK cells using flow cytometry. RESULTS: In the group of patients after IVF failure the percentage of B-1 CD 19+5+ lymphocytes was higher than 1.5% and significantly higher when compared to controls. Three patients after IVF failure had elevated percentages of peripheral blood NK cells. Fourteen infertile patients after IVF failure (82.3%) had at least one abnormal result on autoimmune testing. One patient from the study group had no positive results of immunological tests. CONCLUSION: Our results suggest that immunological alterations may be involved in the etiopathogenesis of unexplained infertility. Furthermore, the results suggest that there is a need for immunological diagnostics in the group of patients with unexplained infertility A greater number of patients is needed for further investigations.     

Preeclampsia is Associated with Lower Percentages of Regulatory T Cells in Maternal Blood

Objective: Immunological mechanisms are involved in the pathophysiology of preeclampsia. During pregnancy there is an increase in regulatory T (Treg) cells, which has an important role in regulating tolerance to the immunologically distinct fetus. We hypothesised that percentages of Treg cells are decreased in preeclamptic patients. Methods: Peripheral blood was obtained from 26 healthy pregnant controls and 18 preeclamptic patients. Treg cells were measured using flow-cytometry. Results: Women with pregnancies complicated by preeclampsia had significantly lower percentages of CD4⁺FOXP3⁺ Treg cells. Conclusion: We conclude that a deficiency of regulatory T cells may play a role in the pathophysiology of preeclampsia.     

CD4+CDhigh25调节性T淋巴细胞在妊娠肝内胆汁淤积症发病中的作用

目的 探讨妊娠肝内胆汁淤积症(ICP)患者外周血与蜕膜组织CD4+CD25+凋节性T淋巴细胞(Treg细胞)及CD4+CD25hJigh细胞在ICP发病中的作用.方法 采用流式细胞仪检测30例ICP患者(ICP组,其中轻度15例,重度15例)及28例正常晚期妊娠妇女(对照组)外周血和蜕膜组织CD4+CD25+Treg细胞及CD4+CD25hJigh细胞占CD+T淋巴细胞的百分率,分析其与ICP发病的关系.结果 ICP组外周血CD4+CD25+Treg细胞百分率为(7.96±1.32)%,CD4+CD25high细胞了百分率为(0.78±0.22)%,均低于对照组[(17.05±2.86)%、(1.71±0.69)%],分别比较,差异均有统计学意义(P<0.01);ICP组蜕膜组织CD4+CD25+Treg细胞百分率为(17.18±2.27)%,CD+4CD25highTreg细胞百分率为(2.25±0.89)%,也均低于对照组[(32.01±3.88)%、(8.30±1.13)%],分别比较,差异也均有统计学意义(P<0.01).ICP组及对照组蜕膜组织中CD4+CD25+Treg细胞及CD4+CD25highTreg细胞百分率均高于外周血,差异均有统计学意义(P<0.01).ICP组轻度及重度患者外周血CD4+CD25+Treg细胞百分率分别为(8.74±0.96)%、(7.17±1.17)%,CD4+CD25hiithTreg细胞百分率分别为(0.89 ±0.20)%、(0.68±0.19)%,而蜕膜组织CD4+CD25+Treg细胞百分率分别为(18.43 ±1.90)%、(15.94±1.95)%,CD4+CD25highTreg细胞百分率分别为(2.62±0.72)%、(1.87±0.90)%,ICP组外周血及蜕膜组织CD4+CD25high胞及CD4+CD25+Treg细胞百分率随着病情的加重呈下降趋势,分别比较,差异均有统计学意义(P<0.05).结论 CD4+CD25+Treg细胞及CD4+CD25+highTreg细胞可能参与ICP的发病过程,并与病情的严重程度密切相关.     

原因不明复发性流产患者蜕膜CD4~+CD25~+CD127~(dim/-) Treg细胞的表达频率和功能研究

目的:探讨原因不明复发性流产(URSA)患者母胎界面免疫耐受环境的变化。方法:留取URSA患者21例(URSA组)和30例正常早孕人流妇女(对照组)蜕膜组织,制备成单个核细胞悬液,流式细胞术分析CD4+CD25+CD127dim/-Treg细胞的表达频率;取两组蜕膜单个核细胞悬液各5例,用免疫磁珠分离法分离出CD4+CD25+CD127dim/-Treg细胞,流式细胞术分析CD4+CD25+CD127dim/-Treg细胞内IL-10和TGF-β的表达水平;体外增殖抑制试验分别检测用抗IL-10抗体和抗TGF-β抗体处理的CD4+CD25+CD127dim/-Treg细胞,和未用抗体处理的CD4+CD25+CD127dim/-Treg细胞对自身效应T细胞增殖的抑制作用。结果:URSA组蜕膜CD4+CD25+CD127dim/-Treg细胞的表达频率明显低于对照组(P=0.005);URSA组蜕膜CD4+CD25+CD127dim/-Treg细胞内IL-10和TGF-β表达频率均较对照组明显降低(P=0.04,P=0.01);URSA组CD4+CD25+CD127dim/-Treg细胞对自体效应性T细胞的增殖抑制作用显著低于对照组(P0.05),且抗IL-10抗体和抗TGF-β抗体可部分阻断蜕膜CD4+CD25+CD127dim/-Treg细胞的免疫抑制功能(P均0.05)。结论:URSA患者蜕膜CD4+CD25+CD127dim/-Treg细胞不仅表达频率降低,而且免疫抑制功能也减弱,且这种免疫抑制功能的减弱主要受IL-10和TGF-β介导,揭示CD4+CD25+CD127dim/-Treg细胞对URSA患者蜕膜局部免疫耐受环境产生影响。     

Dysregulation of macrophage activation by decidual regulatory T cells in unexplained recurrent miscarriage patients

CD4⁺CD25⁺ T cells (Treg cells) and macrophages play roles in the maintenance of maternal–fetal immunological tolerance. Treg cells suppress the function of macrophages via mechanisms mediated by cell–cell contact and production of soluble factors. The purpose of this study was to investigate regulation of macrophages by Treg cells within decidua from patients with unexplained recurrent miscarriage (RM) and normal control women during early pregnancy. Treg cells and macrophages were isolated from deciduas of unexplained RM (n = 15) and control women (n = 15) by magnetic cell separation and co-cultured for six days. Regulation of macrophages by Treg cells was assessed in the presence and absence of neutralizing anti-TGFβ antibodies and in transwell experiments. Expression of CD80, CD86, IL10, and IFNγ by macrophages was measured by flow cytometry or ELISA. Macrophage expression of CD80 and CD86 was higher in deciduas of unexplained RM patients compared with controls whereas the expression of IL10 was lower. There was no difference in the expression of IFNγ by macrophages between the two groups. Treg cells inhibited macrophage expression of CD80, CD86 and IFNγ and increased the expression of IL10. The regulatory effects of Treg cells were abrogated in the presence of neutralizing anti-TGFβ antibodies or by transwell culture. The phenotype of macrophages therefore differed in unexplained RM patients compared with normal early pregnant subjects. Macrophage regulation by Treg cells was shown to be mediated by cell–cell contact and TGFβ and this capacity was decreased in unexplained RM patients.