Spontaneous and LPS-induced secretion of cytokines by villous chorion tissue

Secretion of IL-1α, IL-1α, IL-6, IL-10, and TNF-α cytokines by villous chorion cultures (7–14 weeks) during normal pregnancy and in spontaneous abortions was studied. Secretion of IL-1α and IL-6 increased 4.5 and 7.3 times in miscarriages, while secretion of IL-1α, IL-10, and TNF-α decreased. LPS stimulated the production of IL-1α and IL-6 in samples obtained during surgical abortion. LPS stimulated IL-1α and TNF-α secretion in miscarriages, while the level of IL-6 production decreased significantly. It is hypothesized that increased production of IL-1α and IL-6 and attenuation of the antiinflammatory effect of IL-10 play an important role in the pathogenesis of miscarriages at early stages of gestation. The results suggest that cytokine regulation of the fetus rejection is different at early and late stages of gestation. __________ Translated from Byulleten” Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 6, pp. 668–671, June, 2006     

孕酮对自然流产小鼠白细胞介素6表达的影响

目的:探讨孕酮对自然流产小鼠母胎界面白细胞介素6(IL-6)的调节作用.方法:建立经典的自然流产动物模型(CBA/J×DBA/2),用免疫组织化学显色和RT-PCR检测自然流产小鼠蜕膜及绒毛组织中IL-6蛋白和mRNA的表达情况.结果:孕酮组血清孕酮水平明显高于模型组,IL-6水平高于模型组.孕酮组血清孕酮、IL-6水平与正常组比较均无差异.免疫组织化学显色和RT-PCR结果显示,在绒毛组织中,孕酮组IL-6蛋白相对含量显著高于模型组,IL-6 mRNA相对含量高于模型组,两者与正常组比较均无差异;在蜕膜组织中,孕酮组IL-6蛋白的相对含量低于模型组,IL-6 mRNA相对含量高于模型组,与正常组比较均无差异.结论:孕酮通过上调IL-6的表达来减少自然流产的发生和发展,而且孕酮调节Th1/Th2细胞因子平衡的作用发生在母胎界面.     

Endometriosis: Secretion of interleukin-6 by human endometriotic cells and regulation by proinflammatory cytokines and sex steroids

Endometriosis is generally associated with an immuno-inflammatoryprocess that takes place in the peritoneal cavity of patients.Interleukin (IL)-6, a multifunctional cytokine involved in numerousimmunological and prolifer-ative processes, has been found athigh concentrations in the peritoneal fluid of endometriosispatients. The purpose of this study was to investigate the abilityof endometriotic cells to produce IL-6 and to assess the regulationof its secretion by proinflammatory cytokines and sex steroids.Cultures of human endometriotic cells were exposed to differentconcentrations of cytokines and sex steroid hormones for varyingperiods of time. IL-6 secretion was measured using an enzyme-linkedimmunosorbent assay. Endometriotic cells spontaneously releasedIL-6 in culture. IL-1 and tumour necrosis factor (TNF)- (0.1–100.0ng/ ml) potentiated IL-6 secretion in a time- and dose-dependentmanner. Interferon- (0.4–400 ng/ml) induced a dose-relatedincrease in IL-6 secretion and showed a synergjstic effect onthat secretion in combination with TNF- (10 ng/ ml). Eitherspontaneous or cytokine-induced IL-6 secretion was inhibitedby progesterone (10^–8–10^–5 M) and danazol(10^–6 M), whereas oestradiol (10^–8–10^–5M) had a limited inhibitory effect. The antiprogestin RU486(l0^–8–10^–4 M) antagonized the inhibitory effectsof progesterone and danazol, but showed agonist action whenused alone. These findings indicate that endometriotic tissuemay actively contribute to the biological changes observed inthe peritoneal fluid of endometriosis patients. They also providenew insights into the mechanisms of action of progesterone andthose of danazol and RU486 used in the treatment of endometriosis.     

Polymorphisms in the interleukin-12/18 genes and recurrent spontaneous abortion

PROBLEM: Interleukin (IL) IL-12/IL-18 are involved in uterine NK cells control of uterine vascular development. Polymorphisms in the IL-12/IL-18 genes could modify the cytokine balance, which might result in an increased susceptibility to recurrent spontaneous abortion (RSA). METHOD OF STUDY: A case-control study was conducted to determine the association between the IL12 (I/D) and IL18 (-607C>A, -137G>C) gene polymorphisms and the risk of RSA in 125 women with RSA and in 136 controls. RESULTS: The frequencies of DD, ID, II for IL-12 were, 25.6%, 52.8% and 21.6% respectively, in patients versus 21.3%, 51.5% and 27.2% respectively in controls; the frequencies of CC, CA, AA genotypes for IL-18 (-607) were, 34.4%, 54.4% and 11.2% respectively in patients versus 30.1%, 58.1% and 11.8% respectively in controls; the frequencies of GG, GC, CC genotypes for IL-18(-137) were 47.2%, 43.2% and 9.6% respectively in patients and 45.6%, 46.3% and 8.1% respectively in controls. CONCLUSION: IL-12B and IL-18 promoter gene polymorphisms were not associated with RSA in our women.     

CD147在不明原因自然流产患者绒毛组织的表达

目的:分析CD147在不明原因自然流产患者绒毛组织中的表达水平,并探讨其生物学功能。方法:收集人早孕期正常和不明原因自然流产妇女的绒毛组织,采用免疫组织化学、RT-PCR和Western blotting法检测CD147的表达,比较CD147在正常早孕与不明原因自然流产妇女绒毛组织的表达差异。体外分离纯化获得正常人早孕期滋养细胞,采用免疫细胞化学验证CD147在滋养细胞的表达,引入抗CD147中和性抗体处理人滋养细胞,采用Brd U增殖实验和Transwell侵袭实验分析滋养细胞增殖和侵袭能力的变化。结果:正常早孕期绒毛组织和滋养细胞高表达CD147,不明原因自然流产绒毛组织CD147的mRNA和蛋白表达水平均低于正常早孕期绒毛组织。抗CD147中和性抗体处理原代人滋养细胞后,滋养细胞的增殖能力未发生明显变化,侵袭能力下降。结论:人绒毛组织CD147的异常低表达可能与不明原因早期妊娠失败相关,CD147通过调控滋养细胞的侵袭能力参与正常妊娠的维持。     

细胞周期调节蛋白在早期自然流产患者绒毛和蜕膜组织中的表达及其与自然流产发生的相关性研究

目的：探讨细胞周期调节蛋白在早期自然流产患者绒毛和蜕膜组织中的表达情况并研究其与自然流产发生的相关性。方法：选取符合研究要求的人工流产女性与自然流产女性各35例,分为人工流产组与自然流产组。利用Western blotting与免疫组织化学法分别检测两组患者绒毛与蜕膜组织中细胞周期调节蛋白的表达情况。结果：人工流产组绒毛与蜕膜组织的细胞周期调节蛋白的表达量为75%~85%,自然流产组的表达量为45%~55%。人工流产组与自然流产组比较有统计学意义(P<0.05)。结论：早期自然流产可能与细胞周期调节蛋白在绒毛与蜕膜组织中的含量相关。     

白细胞介素-2在反复自然流产患者绒毛及蜕膜组织中的表达

目的:研究白细胞介素-2(IL-2)在反复自然流产患者绒毛及蜕膜组织中的表达水平及意义.方法:应用免疫组织化学和图像分析,分别对30例反复自然流产患者(病例组)和30例正常妊娠者(对照组)绒毛和蜕膜组织中IL-2的表达进行定位和半定量分析;应用显微镜观察反复自然流产患者绒毛和蜕膜组织的形态学变化.结果:免疫组织化学显色显示,IL-2蛋白定位于绒毛组织的滋养层细胞和蜕膜组织的蜕膜细胞细胞质内,且病例组的IL-2蛋白表达高于对照组.H-E染色结果显示,病例组的蜕膜组织结构模糊、细胞变性坏死、部分细胞连接消失,血管内皮有缺损;绒毛组织的滋养层变薄,细胞变性坏死,细胞嗜酸性增强,绒毛中轴纤维化程度增强.结论:绒毛和蜕膜组织中IL-2蛋白的高表达与反复自然流产有关,可能参与反复自然流产的病理过程.     

蜕膜组织MMP-9/TIMP-3水平与自然流产关系

目的研究蜕膜组织中MMP-9／TIMP-3之间的平衡与自然流产发生的关系。方法用免疫组化S-P法测定30例自然流产患者与20例正常妊娠者蜕膜组织中MMP-9／TIMP-3的表达。结果研究组蜕膜细胞MMP-9表达阳性率为76．7％，高于对照组(55．0％，P-0．02)，两组蜕膜细胞TIMP-3的表达差异无显著性。结论自然流产患者蜕膜组织中MMP-9的表达增高，TIMP-3表达正常所导致的MMP-9／TIMP-3比例升高，在自然流产的发生中起重要作用。     

复发性自然流产患者绒毛及蜕膜组织中白细胞介素8的表达

目的:研究复发性自然流产患者绒毛及蜕膜组织中白细胞介素8(interleukin-8,IL-8)的表达。方法:应用免疫组化法及图像分析技术,对30例复发性自然流产患者绒毛、蜕膜组织中IL-8的蛋白表达进行定位及半定量分析;H- E染色后光镜下观察绒毛及蜕膜组织的形态学变化。结果:在绒毛组织中,IL-8蛋白定位于绒毛上皮细胞的细胞质内,且病例组的表达明显高于对照组;在蜕膜组织中,病例组蜕膜细胞的胞质内可见IL-8蛋白表达,且高于对照组;H- E染色可见病例组绒毛组织的滋养层变薄,细胞变性甚至坏死、嗜酸性增强,绒毛中轴纤维化程度增强;蜕膜组织中蜕膜细胞失去细胞间连接,部分蜕膜细胞解体、核消失。结论:病例组绒毛及蜕膜组织中IL-8蛋白表达的增强与复发性自然流产有关,IL-8可能参与复发性自然流产的病理过程。     

Circulating cytokines and CD30 in normal human pregnancy and recurrent spontaneous abortions

Concentrations of the T-helper (Th) 1 cytokines interleukin (IL)-2, tumour necrosis factor (TNF) -alpha, TNF-beta and interferon-gamma, Th2 cytokines IL-4, IL-5, IL-6, IL-10 as well as those of soluble CD30 in sera have been examined during the three trimesters of gestation, at delivery in normal pregnancy, and at the time of spontaneous abortion in women with a history of unexplained recurrent spontaneous abortion (RSA). Significantly higher concentrations of the Th2 cytokines IL-6 and IL-10 were found at normal delivery than in women with RSA, and conversely significantly increased concentrations of the Th1-type cytokine TNF-alpha were found in RSA as compared with successful pregnancy. In abortion-prone women who had a successful pregnancy, significantly higher concentrations of IL-6 and significantly lower concentrations of TNF-alpha were found as compared with abortion-prone women who had another abortion, supporting the notion that Th2- and Th1-bias are associated with successful and unsuccessful pregnancy respectively. Serum CD30 concentrations did not correlate with the outcome of pregnancy. These findings support observations drawn from experiments on the cytokine secretion profiles of peripheral blood mononuclear cells and decidual lymphocytes which suggest that normal pregnancy is Th2-biased and that unexplained RSA is associated with Th1-type reactivity.     

Maternal serum levels of pro-inflammatory cytokines in missed and threatened abortion

PROBLEM: A study of association between pro-inflammatory cytokines, and missed and threatened abortions with good outcome has been performed. METHOD OF STUDY: The presence of pro-inflammatory cytokines, namely interleukin (IL)-8 and IL-12 and the soluble interleukin-2 receptor (SIL-2R) was investigated in maternal serum of 12 patients with threatened abortion twice (at admission and discharge), 14 patients with missed abortion, 14 women with healthy first-trimester pregnancy, and 14 normal non-pregnant women, using specific enzyme-linked immunosorbent assays. RESULTS: SIL-2R and, in particular, IL-12 was detected with significantly higher levels in missed abortion group compared with all other groups. IL-8 was detected with no significant difference among all the groups studied. CONCLUSIONS: In spite of caution due to the small sizes of the subject samples, these results support a role of the immune system in the first trimester pregnancy and hypothesize that missed abortion may be associated with an enhanced Th1 reactivity, whereas threatened abortion with good outcome resembles the normal pregnancy with a non-enhanced Th1 reactivity.     

Prevention of recurrent spontaneous abortion by intravenous immunoglobulin: a double-blind placebo-controlled study

The aim of this study was to evaluate the therapeutic efficacy of intravenous immunoglobulin (IVIG) in the prevention of recurrent spontaneous abortion (RSA). In a double-blind, randomized, placebo-controlled study, 41 women with a history of unexplained recurrent spontaneous abortion were treated with IVIG or saline infusions during pregnancy. The birth of a child was considered a successful outcome. The overall success rate was 77% in the IVIG group compared with 79% in the placebo group. For women with primary RSA the success rates were 82 (IVIG) and 89% (placebo), and for women with secondary RSA the rates were 73 (IVIG) and 70% (placebo). We found no statistically significant difference in treatment results between IVIG and placebo.     

HLA-E polymorphism in patients with recurrent spontaneous abortion

Abstract: The aim of this study was to compare the frequencies of five HLA-E alleles in 82 women with recurrent spontaneous abortions with that of 150 random Danish controls. PCR sequence-specific oligonucleotide typing was carried out to detect polymorphism in exons 2 and 3 of the HLA-E gene. In informative samples sequencing of these two exons was also undertaken to confirm the presence of the HLA-E*01031 allele. HLA-E*0101, HLA- E*01032 and HLA-E*01031 were detected with frequencies of 56.7%, 33.6% and 9.6% in controls and 58,5%, 32.9% and 8.5% in patients with recurrent abortion, respectively. No HLA-E*0102 and E*0104 alleles could be detected. Since the HLA-E allele distribution was similar in women with recurrent spontaneous abortion and controls, it is suggested that maternal HLA-E polymorphism per se does not play any role in the pathogenesis of this disorder of pregnancy.     

复发性流产的免疫治疗效果分析

目的了解采用淋巴细胞对复发性流产进行主动免疫的疗效。方法采用丈夫外周血淋巴细胞治疗复发性流产,454例患者疗程结束并成功随访,妊娠至20周后者为治愈。结果116（25．55％）例足月分娩,治愈率70．48％（未包括已孕20周内）。其间1例孕20周诊断无脑儿,1例孕26周超声示胎儿先天性左心发育不良,畸形率0．44％。结论免疫治疗是安全、有效的治疗复发性流产的方法之一,能确实解决病患实际问题。     

Epidemiology of recurrent spontaneous abortion.

With recent scientific advances leading to better understanding of the immunobiology of recurrent spontaneous abortion (RSA), interest has now focused upon the epidemiology of RSA. A cohort of 214 couples with a history of two or more consecutive abortions were studied for the prevalence of etiologic factors and association with other reproductive failures. The prevalence of causes of RSA in this cohort was compared with etiologic factors among 179 couples with a history of three or more consecutive abortions. The obstetrical histories of 214 women with RSA were analyzed for the total number of pregnancies, live births, stillbirths, spontaneous abortions, ectopic pregnancies, and hydatidiform moles. These numbers were compared with the expected frequency of each in the general population. The prevalence of etiologies among 214 with RSA were as follows: chromosomal-6%, anatomic-1%, hormonal-5%, immunologic-65%, and unexplained-23%. No differences in the prevalence of etiologic factors exist when couples with a history of two or more abortions are compared with three or more abortions. When the number of ectopic pregnancies, molar pregnancies, and stillbirths among 214 women with RSA were compared with the expected numbers, the odds ratios were 2.2 for ectopic pregnancies, 6.0 for molar pregnancies, and 2.3 for stillbirths. These data indicate that no difference in the prevalence of etiologies of RSA exist when couples with two or more abortions are compared with three or more and a comorbidity between RSA and other types of reproductive failure exists.     

主动免疫治疗复发性自然流产的研究

目的探讨主动免疫在治疗复发性自然流产中的作用。方法选择年龄24～40岁、原因不明性复发性自然流产、封闭抗体阴性患者26例为试验组,选择同期在我科就诊、年龄相当、原因不明性复发性自然流产、封闭抗体阴性患者24例为对照组。对试验组进行淋巴细胞注射及常规保胎治疗,对照组使用常规保胎治疗。结果试验组妊娠成功率为73%,对照组为21%,P〈0.05,差异有显著性。结论主动免疫疗法在治疗原因不明性复发性自然流产中疗效确切,具有操作有效、无副作用等特点。     

自然流产患者蜕膜组织NK细胞受体表达分析

目的研究自然流产患者子宫蜕膜组织中NK细胞受体的表达格局。方法采用RT-PCR和免疫组织化学法检测了12例早期自然流产及40例同期正常早孕要求行人工流产者蜕膜组织的NK细胞受体表达水平。结果NK细胞受体中阳性最高的是KIR2DL4,流产组和对照组分别为100%和95%,其次是KIR2DL1,分别为83.3%和75%。ILT2和ILT4的阳性率较低,流产组和对照组分别为25%、30%和8.3%、10%。统计结果显示,KIR2DL4、KIR2DL1、ILT2和ILT4mRNA阳性率在早期自然流产病人组和对照组之间均无显著性差异。与对照组相比,早期自然流产病人的蜕膜组织中KIR2DL4分子的表达水平明显降低,具体表现在KIR2DL4的分布密度上。结论早期流产的发生与NK细胞受体的转录水平可能并无直接关联,但KIR2DL4分子的表达水平高低,可能对早期胚胎的生长、发育起关键作用。     

复发性流产患者抗精子抗体检测与分析

目的探讨抗精子抗体在复发性流产患者孕前检查中的应用价值。方法选取420例有复发性流产史患者作为试验组,其中男189例,女231例,对其进行抗精子抗体检测;同时选取有正常生育史的45例男性与65例女性作为参照组,对两组的抗精子抗体的检测结果进行比较分析。结果试验组女性的阳性率为10.39%,男性的阳性率为7.26%,对照组女性的阳性率为1.54%,男性对照组中无人发现抗体阳性,RSA组的抗体阳性率明显高于正常对照组,差别有统计学意义（P〈0.05）。结论 As Ab的产生与RSA密切相关,在对复发性流产患者的优生咨询中,需重视免疫学因素。     

封闭抗体与复发性自然流产的研究进展

复发性自然流产(RSA)是妊娠期妇女常见的并发症,在育龄妇女中发病较常见,导致RSA的原因复杂主要涉及到染色体、解剖、内分泌、感染、自身抗体等多方面因素,其中大多数与免疫因素有关.近年来封闭抗体(APLA)与RSA之间的关系得到了越来越多研究者的关注.APLA缺乏是导致RSA的免疫学因素之一,因而具有重要的临床意义.