地屈孕酮对不明原因复发性自然流产患者外周血辅助T细胞的影响

目的探讨地屈孕酮补充治疗对不明原因复发性自然流产(URSA)患者外周血辅助性T细胞(Th)亚群Th1/Th2的影响。方法选取30例不明原因复发性自然流产患者为URSA组,另选取30例正常育龄妇女作为对照组,URSA组予以地屈孕酮治疗,对照组不采取任何治疗措施。两组均进行γ干扰素(IFN-γ)(Th1细胞因子)、白细胞介素-4(IL-4)(Th2细胞因子)的检测,观察URSA组妊娠情况,比较两组Th细胞因子变化情况、妊娠后不同时期Th1/Th2的变化,分析地屈孕酮对不同流产次数患者再次妊娠成功后Th细胞因子的影响。结果30例URSA患者妊娠28例,其中2例生化妊娠,3例妊娠40+d胚胎停育清宫,23例妊娠成功已转入产科定期产前检查。URSA组外周血Th1型细胞因子IFN-γ(26.37±3.13)pg/ml高于对照组的(17.01±3.22)pg/ml,差异有统计学意义(P<0.05)。两组Th2型细胞因子IL-4比较差异无统计学意义(P>0.05)。流产2次患者与流产>2次患者之间,IFN-γ活性出现升高趋势,Th1/Th2比值明显向Th1偏移。妊娠40 d及妊娠3个月,外周血Th1型细胞因子IFN-γ活性低于妊娠前,差异有统计学意义(P<0.05);与对照组比较差异无统计学意义(P>0.05);Th2型细胞因子IL-4活性高于妊娠前,差异有统计学意义(P<0.05)。Th1/Th2模式向后者偏移,Th1/Th2比值为4.37 vs 2.37。至孕3个月,偏移越趋于明显,5例妊娠失败患者外周血中,Th1型细胞因子IFN-γ及Th2型细胞因子IL-4与妊娠前比较,差异无统计学意义(P>0.05)。经地屈孕酮治疗后,流产2次的URSA患者Th1型细胞因子IFN-γ(16.03±5.11)pg/ml低于流产>2次的(21.48±1.83)pg/ml,差异有统计学意义(P<0.05);流产>2次的URSA患者呈现出更为明显的Th2偏移现象。结论地屈孕酮治疗可以使Th1/Th2平衡向Th2偏移,对免疫调节起重要作用,有利于妊娠成功。     

Gonadotrophin surge-attenuating factor bioactivity is present in follicular fluid from naturally cycling women

Rat pituitary monolayer bioassays were used to compare gonadotrophinsurge-attenuating factor (GnSAF) bioactivity in follicular fluidfrom 12 follicles in 10 spontaneously cycling women with thatin pooled follicular fluid from women undergoing ovulation induction.Expressed as ED₅₀s (µl follicular fluid/well producing50% of maximal effect), GnSAF bioactivity was detectable inall spontaneous follicular fluid samples (1.4–33.3 µl/well)and in follicular fluid from women undergoing ovulation induction(6.8 µl/well). This GnSAF bioactivity was unaffected bypre-incubation with an inhibin antibody. When the data weregrouped according to whether the recovered oocytes fertilizedin vitro or not, the fertilized group contained significantlygreater GnSAF bioactivity than the unfertilized group (5.3 ±1.1 and 14.1 ± 2.6 µl/well respectively, P <0.05). While both inhibin bioactivity (9.7 ± 1.4 and28.9 ± 12.1 µl/well) and immunoreactivity (36.8± 2.2 and 21.0 ± 3.0 and ng/ml) were also greater(P < 0.01) in the fertilized compared with the unfertilizedgroups respectively, there were no other significant differencesbetween the two groups. We conclude that GnSAF is found in follicularfluid from spontaneously cycling women, supporting in-vivo evidencefor the involvement of GnSAF in feedback control of the ovary-pituitaryaxis.     

反复自然流产的免疫因素分析探讨

为分析探讨反复自然流产的免疫性因素，应用补体微量淋巴细胞毒实验方法检测女性反复自然流产的封闭抗体，用ELISA法检测女性反复自然流产的抗心磷脂抗体(ACA)、抗精子抗体(ASA)共752例。结果显示，封闭抗体与反复自然流产明显相关，抗心磷脂抗体与反复自然流产有一定关系，抗精子抗体与反复自然流产无明显相关。研究表明，封闭抗体、抗心磷脂抗体是造成反复自然流产的重要免疫因素。     

Prevention of Unexplained Recurrent Spontaneous Abortion Using Intravenous Immunoglobulin: A Prospective,Randomized, Double-Blinded,Placebo-Controlled Trial

PROBLEM: The efficacy of intravenous immunoglobulin (IVIG) for treatment of unexplained recurrent spontaneous abortion was assessed in a prospective, randomized, double-blinded, and placebo-controlled study. METHOD OF STUDY: The study took place in a provincial recurrent pregnancy loss clinic, located in a tertiary/quaternary care academic center. The study subjects were women with a history of two or more documented consecutive spontaneous pregnancy losses under 20 weeks of gestation, excluding any associated with aneuploidy by karyotype analysis, and with no evidence of genetic, endocrine, infectious, anatomic, or autoimmune factors associated with a history of recurrent spontaneous abortion. The subjects were randomized to receive either intravenous immunoglobulin (Gamimune N) as treatment or normal saline as placebo. Randomization was stratified for primary, secondary, and unclassified unexplained recurrent spontaneous abortion. Success was defined as an ongoing pregnancy beyond 20 weeks of gestation. RESULTS: Sixty-two subjects enrolled in the trial. There were 37 index pregnancies and 6 cross-over pregnancies. There was no clinically significant difference between the treatment arm and the placebo arm in terms of subsequent pregnancy success. There seemed to be a higher success rate with the stratified analysis of couples with secondary unexplained recurrent spontaneous abortion, but the trial did not have sufficient power to confirm this. CONCLUSIONS: Based on this trial and three similar trials in the literature, a multicentered trial is needed to determine conclusively whether IVIG is effective in the treatment of unexplained recurrent spontaneous abortion.     

配偶淋巴细胞免疫治疗反复自然流产91例分析

目的探讨反复自然流产患者,运用配偶淋巴细胞免疫治疗后,体内封闭抗体水平的变化及其临床疗效.方法对确诊为封闭抗体缺乏的RSA患者91例,用分离的配偶淋巴细胞,皮内注射法免疫治疗3～5疗程,治疗前后运用流式细胞仪测定患者体内封闭抗体水平.结果 91例RSA患者经免疫治疗后,体内封闭抗体的水平显著提高,抗配偶CD3%、抗配偶CD4%、抗配偶CD8%,分别由治疗前的-1.32±0.98、-1.39±0.80、-0.96±0.54上升为0.99±0.74、1.01±0.64、0.75±0.49(P＜0.05);妊娠成功80例,成功率为87.9%(80 / 91).结论配偶淋巴细胞免疫治疗,可以提高患者体内封闭抗体水平,有助于妊娠成功.     

活动性巨细胞病毒感染与反复自然流产的关系探讨

目的探讨活动性人巨细胞病毒（human cytomegalovirus,HCMV）感染与反复自然流产（recurrent spontaneous abortion,RSA）的关系.方法采集反复自然流产孕妇和正常产前体检孕妇外周血,分离外周血单个核细胞（PBMCs）和血浆,分别用免疫荧光法和实时定量PCR检测HCMV pp65抗原和HCMV DNA,并比较2种方法的一致性.结果 65例RSA患者HCMV pp65抗原有20例阳性,阳性率30.8%,50例正常体检孕妇 HCMV pp65抗原有4例阳性,阳性率8.0%,2组孕妇HCMV活动性感染率有显著性差异（χ^2=8.87,P＜0.01）.孕妇HCMV pp65抗原阳性率升高,孕妇流产几率增加（χ^2=7.53,P＜0.01）. 免疫荧光法和实时定量PCR有较好的一致性（92.3%）.结论反复自然流产孕妇 HCMV活动性感染率显著高于正常孕妇,HCMV pp65抗原检测也许可作为RSA早期诊断指标之一.     

Immunotherapy for Recurrent Pregnancy Loss: Analysis of Results From Clinical Trials

PROBLEM: Up to 80% of unexplained recurrent spontaneous abortions (RS A) are thought to have an immunologic mechanism. Yet clinical trials using immunotherapy to treat women experiencing RSA have low treatment effects. The present study was undertaken to explain the low treatment effects. METHODS: Results of clinical trials using allogeneic leukocyte immunization and intravenous (IV) immunoglobulin (Ig) are compared. The mechanisms of pregnancy loss are reviewed in light of data on frequency of karyotype abnormalities in trophoblast of failing pregnancies. RESULTS: Results of two independent analyses using allogeneic leukocyte immunization as immunotherapy for all women with RSA revealed live birth ratios of 1.16 (P = 0.03) and 1.21 (P = 0.02). When the analysis was limited to primary aborters, the live birth ratio increased to 1.46 (P = 0.006). Live birth ratio after immunotherapy for all RSA using IVIg was 1.88 (P = 0.04). Because of low treatment effects, confounders to treatment success of maternal age and number of previous abortions were studied. Chromosomal abnormalities have been identified in 55% of concepti from RSA. The frequency of chromosomal abnormalities remained constant for up to six pregnancy losses. Women with a history of primary compared to secondary RSA had a higher frequency of karyotypically abnormal concepti (χ² = 4.54, P < 0.05). Risk factors for RSA also include number of previous losses. CONCLUSION: Chromosomal abnormalities are a significant confounder when evaluating efficacy of immunotherapy for treatment of RSA. Some women with RSA have a high risk of recurrent chromosomal problems.     

Allo-immunization elicits CCR5 antibodies,SDF-1 chemokines,and CD8-suppressor factors that inhibit transmission of R5 and X4 HIV-1 in women

Studies in humans suggest that allo-immunization induces CC-chemokines, CD8-suppressor factors (SF) and anti-HIV immunity. Here we report that allo-immunization with unmatched leucocytes from partners of women with recurrent spontaneous abortion elicits specific antibodies to the CCR5 receptor. Such antibodies inhibit replication of M-tropic HIV-1 (R5) and MIP-1beta-mediated chemotaxis. These CCR5 antibodies were also found in the sera of multiparous women that were naturally immunized by semi-allogeneic fetal antigens. The specificity of these antibodies was demonstrated by adsorption with CCR5 transfected HEK-293 cells, a baculovirus CCR5 preparation and a peptide of the 2nd extra-cellular loop of CCR5. Allo-immunization also stimulated increased concentrations of the CXC chemokine, SDF-1alpha and CD8-SF that inhibit T-tropic HIV-1 (X4) replication. We suggest that allo- immunization may elicit (a) CC chemokines, CCR5 antibodies and CD8-SF that inhibit M-tropic HIV-1 infection and (b) the CXC chemokine SDF-1alpha and CD8-SF that inhibit T-tropic HIV-1 infection.     

Preventive effect of monoclonal antibodies to intercellular adhesion molecule-1 and leukocyte function-associate antigen-1 on murine spontaneous fetal resorption

PROBLEM: Are cell adhesion molecules involved in the murine model of immunologically‐mediated spontaneous abortion?
METHOD OF STUDY: Pregnant CBA/J female mice mated with DBA/2 male mice were injected with monoclonal antibodies (MAbs) to intercellular adhesion molecule‐1 (ICAM‐1) and leukocyte function‐associate antigen‐1 (LFA‐1). On day 13 of gestation, viable and resorbed embryos were counted. Natural killer (NK) cell activity in the spleen, mixed lymphocyte reactions (MLR), mixed lymphocyte‐placenta reactions (MLPR), and levels of interferon (IFN)‐Γ were assayed.
RESULTS: Significant suppression of fetal resorption was observed by the injection of MAb to ICAM‐1 and LFA‐1. NK cell activity and the MLR anti‐(CBA/J×DBA/2)F1 were reduced in the antibody‐treated CBA/J spleen. Moreover, the level of IFN‐Γ was significantly lower in the MLPR supernatants from the antibody‐treated group than those of the control group.
CONCLUSIONS: One mechanism in the murine model of spontaneous abortion may be through the interaction of cell adhesion molecules, which may modulate NK cell activities and cytokine production.