免疫治疗原因不明习惯性流产后外周血T淋巴细胞亚群的变化

目的:探讨原因不明习惯性流产患者外周血T淋巴细胞亚群在主动和被动免疫治疗前后的变化。方法:采用流式细胞仪对2 5例被动免疫治疗和38例主动免疫治疗的原因不明性习惯性流产患者作治疗前后外周血T淋巴细胞亚群含量对比,并选择2 0例正常妊娠者作对照组。结果:①被动与主动免疫组治疗前后CD3+细胞无明显差异。②被动和主动组治疗后CD4 +细胞较治疗前有明显减少。③CD8+细胞在被动免疫治疗和主动免疫治疗后较治疗前有明显升高。④CD4 +/CD8+比率在被动免疫治疗和主动免疫治疗后较治疗前明显下降。结论:主动和被动免疫治疗后,外周血T淋巴细胞亚群有了明显变化,更利于妊娠发展。     

淋巴细胞主动免疫治疗对原因不明复发性流产机体免疫功能的影响

目的探讨淋巴细胞主动免疫治疗对原因不明复发性流产患者机体体液免疫和细胞免疫的影响。方法对2008年1月至2009年1月广东省计划生育专科医院收治的89例复发性流产患者进行丈夫或第三者淋巴细胞主动免疫治疗,ELISA方法分析其诱导的抗HLA-Ⅰ、Ⅱ抗体水平,流式细胞术分析其中21例妊娠成功者和9例妊娠失败者体内IFN-γ+CD4+T、IL-10+CD4+T和NK细胞亚群频率变化。结果 89例患者经主动免疫治疗后,有72例妊娠,其中63例成功妊娠,妊娠成功率为87.50%;妊娠成功者中共有61例抗HLA-Ⅰ和(或)抗HLA-Ⅱ抗体转阳,总转阳率为96.83%。9例妊娠失败者中仅有1例抗HLA-Ⅰ抗体转阳,总转阳率为11.11%。妊娠成功者体内IFN-γ+CD4+T和CD56+CD16+NK细胞频率明显下降,而IL-10+CD4+T细胞频率明显升高;而妊娠失败者体内各细胞亚群无显著性改变。结论淋巴细胞主动免疫治疗能显著下调机体细胞免疫应答,上调机体体液免疫应答,更利于妊娠发展。     

不明原因复发性流产主动免疫治疗前后封闭抗体的变化及其意义

目的:探讨淋巴细胞主动免疫治疗不明原因复发性流产患者的疗效及治疗前后封闭抗体(BA)变化及对再次妊娠的影响.方法:对81例BA阴性的不明原因复发性流产患者进行淋巴细胞主动免疫治疗,观察其治疗后BA的变化及再次妊娠的结局.结果:在81例治疗的患者中,有73例获妊娠成功,8例再次发生早期流产,妊娠成功率90.12%(73/81);淋巴细胞主动免疫后43例复查,其中35例BA转阳性,阳性率81.40%(35/43),BA阳性率与治疗前比较,差异有高度统计学意义(P<0.01);35例BA转阳性者均妊娠成功,BA仍阴性者8例,继续进行3次主动免疫治疗后5例妊娠成功,有3例再次发生早期流产,妊娠成功率BA阳性者高于阴性者,差异有统计学意义(P<0.05).结论:淋巴细胞主动免疫治疗能提高BA阴性的不明原因复发性流产患者的再次妊娠成功率及提高BA的阳性率,对不明原因复发性流产患者再次妊娠的结局有一定的参考价值.     

反复胚胎种植失败免疫治疗疗效及淋巴细胞免疫表型变化的研究

目的:探讨淋巴细胞主动免疫治疗不明原因反复胚胎种植失败(RIF)患者的疗效及治疗前后外周血淋巴细胞免疫表型的变化。方法:对研究组83例不明原因RIF的患者进行淋巴细胞主动免疫治疗(LIT);对照组96例不明原因RIF患者移植前不给予特殊处理。并按照胚胎移植属性分为鲜胚移植和冻胚移植,所移植胚胎均为卵裂期优质胚胎,接受LIT后新鲜胚胎移植为研究组A(51例);未行LIT接受新鲜胚胎移植为对照组A(57例)。接受LIT后冷冻胚胎移植为研究组B(32例),未行LIT接受冷冻胚胎移植为对照组B(39例)。比较各组的临床妊娠情况,并观察LIT治疗前后外周血淋巴细胞免疫表型变化。结果:(1)研究组A的临床妊娠率(47.06%)高于对照组A(22.81%),研究组B的临床妊娠率(46.88%)高于对照组B(23.77%),差异有统计学意义(P0.05);研究组A的临床妊娠率与研究组B比较,差异无统计学意义(P0.05)。(2)与治疗前相比,LIT后外周血CD3~+CD4~+T细胞占淋巴细胞的比率下降,CD3~+CD8~+T细胞占淋巴细胞的比率上升、CD4~+/CD8~+比值下降,NK细胞(CD3-CD16~+CD56~+)占淋巴细胞的比率下降,差异均有统计学意义(P0.05)。结论:淋巴细胞主动免疫治疗对RIF患者成功妊娠有促进作用,淋巴细胞主动免疫治疗可能促使外周血淋巴细胞免疫表型向增强母胎免疫耐受的方向改变。     

主动免疫治疗复发性流产妊娠结局与调节性T细胞关系初探

目的:探讨主动免疫治疗原因不明复发性流产患者妊娠结局与调节性T细胞(CD4+CD25+Treg)的关系.方法:采用流式细胞仪测定主动免疫治疗前后复发性流产患者外周血CD4+CD25+Treg比例.结果:主动免疫治疗后CD4+CD25+Treg 的表达较治疗前增高(P＜0.05),且治疗后妊娠成功组与治疗后妊娠失败组比较,差异有统计学意义(P＜0.05).结论:主动免疫治疗可上调CD4+CD25+Treg的表达,可能与妊娠结局有一定相关性.     

自发性早产母胎界面子宫自然杀伤细胞亚群及T淋巴细胞亚群变化的研究

目的:研究不明原因早产发生时,母胎界面中自然杀伤细胞亚群及T淋巴细胞亚群的变化,探讨早产发生机制.方法:采用流式细胞技术,分别检测足月妊娠分娩孕妇30例(正常妊娠组)和不明原因孕周满28周至不满37周无医学指针终止妊娠的孕妇30例(早产组)母胎界面底蜕膜组织中CD4+T细胞、CD8+T细胞、CD56+CD16+NK细胞百分比、Th1及Th2细胞的百分比.结果:①早产组患者CD4+T细胞、CD56+CD16+NK细胞百分比明显高于正常妊娠组(P<0.05),CD8+T细胞百分比明显低于正常妊娠组(P<0.05);②早产组Th1细胞、Th1/Th2比例明显高于正常妊娠组(P<0.01).结论:母胎界面局部T淋巴细胞亚群、子宫自然杀伤细胞百分比的改变及Th11细胞增多与不明原因自发性早产相关.     

主动免疫治疗对原因不明复发性流产患者外周血淋巴细胞Fas表达的影响

目的:探讨经主动免疫治疗后对原因不明复发性流产(URSA)患者外周血淋巴细胞Fas表达的影响。方法:收集来我院就诊URSA患者30例,采用荧光标记流式细胞分析技术,检测经淋巴细胞主动免疫治疗前、一疗程结束后3周的外周血淋巴细胞Fas表达。同时选择同期正常育龄妇女20例,作为正常对照组,比较两组Fas表达情况。结果:①CD4+ T淋巴细胞的Fas表达,正常对照组与URSA组治疗前、后的比较中,差异无统计学意义(P>0.05);②URSA组治疗前的CD8+ T淋巴细胞和NK细胞的Fas表达均比正常对照组明显升高(P<0.05);③URSA组治疗后的CD8+ T淋巴细胞和NK细胞的Fas表达均较治疗前降低(P<0.05)。结论:URSA患者存在CD8+ T淋巴细胞和NK细胞的异常激活,主动免疫治疗可以降低Fas在淋巴细胞上的表达,从而有利于胚胎的发育。     

原因不明复发性流产免疫治疗后局部皮肤反应与调节性T细胞的关系

目的探讨淋巴细胞免疫治疗原因不明复发性流产(URSA)后局部皮肤反应面积与调节性T细胞变化的关系。方法对2008年1月至2009年6月广东省计划生育专科医院收治的40例URSA患者,采用丈夫或无关第三个体淋巴细胞免疫治疗,根据妊娠结局将研究对象分为妊娠成功组和妊娠失败组,分别记录首次和末次免疫治疗后24h和48h皮肤反应面积;应用流式细胞术分析各组首次与末次免疫1周后患者外周血CD4+CD25+调节性T细胞亚群频率变化。结果 40例URSA患者接受免疫治疗后有38例妊娠,其中33例妊娠成功,5例妊娠再次失败。妊娠成功组末次免疫后CD4+CD25brightT细胞频率较首次免疫后明显上调;CD4+CD25dimT细胞频率较首次免疫后明显下调;末次免疫后局部皮肤反应面积较首次免疫后明显缩小;局部皮肤缩小与CD4+CD25brightT细胞频率上升呈负相关;与CD4+CD25dimT细胞频率降低呈正相关。妊娠失败组首次与末次免疫后CD4+CD25brightT、CD4+CD25dimT细胞频率及局部皮肤反应均无显著性改变。结论免疫治疗URSA后局部皮肤反应面积缩小可能与CD4+CD25brightT细胞频率上调、C...     

原因不明性习惯性流产患者外周血T淋巴细胞亚群的变化

采用流式细胞仪对２０例原因不明性习惯性流产（ＵＨＡ组）患者外周血Ｔ淋巴细胞亚群进行检测，同时观察Ｔ淋巴细胞亚群在主动免疫治疗前后的变化，并以２０例正常非妊娠妇女（ＮＮＰ组）、１０例正常妊娠期妇女（ＮＰ组）作为对照。结果：ＵＨＡ组主动免疫治疗前分化群（ｃｌｕｓｔｅｒｏｆｄｉｆｆｅｒｅｎｔｉａｔｉｏｎＣＤ）ＣＤ＿３细胞百分率与ＮＮＰ组相比，差异无显著性；ＣＤ＿８细胞的百分率明显低于ＮＮＰ组（Ｐ＜０．００１），而ＣＤ＿４／ＣＤ＿８比率则明显高于ＮＮＰ组（Ｐ＜０．０５）；主动免疫治疗后，ＣＤ＿８细胞的百分率较治疗前明显上升，ＣＤ＿４／ＣＤ＿８比率有明显下降（Ｐ＜０．０５），而且治疗后ＵＨＡ组妊娠时ＣＤ，细胞的百分比和ＣＤ＿４／ＣＤ＿８比率均与ＮＰ组接近。提示：原因不明性习惯性流产的免疫发病机理可能与ＣＤ＿８细胞数量的下降、ＣＤ＿４／ＣＤ＿８比率的上升有关；主动免疫疗法可以诱导ＣＤ＿８细胞增殖；ＣＤ＿４／ＣＤ＿８比率可作为主动免疫治疗临床疗效的监测指标。     

细胞免疫功能的变化与原因不明习惯性流产的关系

目的探讨细胞免疫功能的变化与原因不明习惯性流产（unexplained habitual abortion，UHA）的发生及主动免疫治疗机制的关系。方法2002-08—2004-08汕头大学医学院第一附属医院采用流式细胞仪测定并比较112例UHA患者和30例正常已生育妇女及76例经主动免疫治疗后的UHA患者外周血CD3^＋、CD4^＋、CD8^＋、CD18^＋CD56^＋细胞亚群比例及CD4^＋／CD8^＋的比值；同时比较76例采用主动免疫治疗和36例未采用主动免疫治疗的UHA者再次妊娠成功率。结果UHA患者与正常已生育妇女比较，血中CD3^＋、CD16^＋CD56^＋细胞的百分率及CD4^＋／CD8^＋比值增高，差异有显著性意义（P〈0．05）。主动免疫后CD3^＋、CDl6^＋CD56^＋细胞百分率及CD4^＋／CD8^＋比值下降（P〈0．05）。经主动免疫治疗的UHA者再次妊娠成功率为88．24％，对照组为31．25％，两组比较差异有非常显著性意义（P〈0．01）。结论淋巴细胞亚群比例的改变与UHA的发生有关，主动免疫治疗可调节异常的细胞免疫功能，有利于提高再次妊娠成功率。     

不明原因习惯性流产患者蜕膜自然杀伤细胞活性及表型的研究

目的 :探讨原因不明习惯性流产 (UHA)患者蜕膜自然杀伤 (NK)细胞的数量、表型特征及杀伤活性 ,探讨其在UHA发病中的作用。方法 :选取确定妊娠并难免流产的UHA 32例 ,选取 2 0例正常妊娠行人工流产者作为对照组 ,用流式细胞仪检测两组蜕膜组织中NK细胞数量及表型 ,用改进的乳酸脱氢酶释放法测定NK细胞杀伤活性。结果 :正常早孕蜕膜中CD5 6+CD3- NK细胞占蜕膜单个核细胞的 60 .73± 13.2 4 % ,为蜕膜组织的优势细胞群 ,UHA患者蜕膜中CD5 6+CD3- NK细胞总量与正常妊娠蜕膜组织中相同 ,而其CD5 6+CD16- 亚群却明显低于正常妊娠组 (P <0 .0 5 ) ,CD5 6+CD16+亚群含量及CD5 6+CD16+/CD5 6+CD16- 比值均异常增高 (P均 <0 .0 5 )。UHA组蜕膜组织中NK细胞杀伤活性及其异常增高率均高于正常妊娠组 ,差异均有显著性 (P均 <0 .0 5 )。结论 :NK细胞是孕早期蜕膜组织中的优势细胞群 ,其亚群失衡及功能异常可导致自然流产     

The evaluation of expression of T-lymphocytes and NK cell surface receptors in women with threatened miscarriage

OBJECTIVES: Recent studies emphasise an important role of immunological mechanisms in pregnancy maintenance. Therefore, unravelling mechanisms regulating placentogenesis are critical to understanding the pathogenesis of recurrent spontaneous abortion. MATERIALS AND METHODS: We studied 49 women with threatened abortion and 24 healthy pregnant women. In addition, we studied 17 women with the history of recurrent spontaneous abortion and 10 healthy nonpregnant women in reproductive age with the previous successful pregnancy outcome. CD3, CD45 RO, CD4, CD8, CD16 expression on peripheral blood lymphocytes were studied using flow cytometry. RESULTS: We determined that there is significantly higher CD4 expression in pregnant recurrent aborters compared to control (p < 0.05). Pregnant recurrent spontaneous aborters, with the successful pregnancy outcome have significantly lower CD16 expression compared to those, who abort (p < 0.05). CONCLUSION: Our studies may indicate that T cell and NK cell can be involved in the pathogenesis of pregnancy loss.     

Immunological evaluation of patients with recurrent abortion

In a prospective study, we performed immunological tests in patients with recurrent abortion. Nine couples with two or more fetal losses of no apparent cause were selected as the patient group, and nine volunteer couples with at least two children and without a history of abortion were used as controls. The frequency of major histocompatibility complex (human leukocyte antigens, HLA) antigen sharing was determined by serological methods, antipaternal antibodies by microlymphocytotoxicity, lymphocyte phenotypes (CD4, CD8, CD19, CD16, CD56 and HLA-DR positive cells) by flow cytometry and natural killer (NK) cytotoxicity by (51)Cr release. NK activity was correlated to the degree of HLA-C sharing and to the percentage of CD16+ and CD56+ cells and to progesterone levels measured by radioimmunoassay. No difference in class I or class II HLA antigen sharing was detected between couples with and without recurrent abortion. Antipaternal antibodies were not found in the serum of any woman of the study. A higher absolute number of CD8+ cells (P=0.01) and a trend to increased CD19+ cells (P=0.05) were observed among patients. NK activity did not differ between the two groups when expressed as specific cytotoxicity and it was reduced among patients with recurrent abortion when expressed as lytic units/10(7) cells (P=0.04). There was correlation between NK activity and the percentage of CD16+ and CD56+ cells but not with progesterone levels in patients with recurrent abortion. Our data suggest that an increased NK activity may not play a role in the occurrence of repeated abortion. On the other hand, an increase in circulating CD8+ T cells was observed in patients suggesting that antifetal cytotoxicity in recurrent abortion may be mediated by T cells and not by NK cells.     

The influence of paternal lymphocyte immunization on the balance of Th1/Th2 type reactivity in women with unexplained recurrent spontaneous abortion

OBJECTIVES: Paternal lymphocyte immunization has been proposed as an efficient treatment for unexplained recurrent spontaneous abortion (RSA), however precise mechanism that underlie the benefits of this immunotherapy are still unclear. It was proposed that successful pregnancy is reminiscent of T helper 2 (Th2)--dominant situation but unsuccessful pregnancy is a Th1-type situation. The aim of the study was the evaluation of influence of paternal lymphocyte immunization on the balance of Th1/Th2--type reactivity in women with unexplained recurrent spontaneous abortion. MATERIAL AND METHODS: 8 patients with a history of 3 or more consecutive primary spontaneous abortions of unknown etiology and no positive autoimmune factors were selected for the study. Immunization with paternal lymphocytes, obtained from 100 ml of peripheral blood, was performed twice prior conception with a 4-week interval. The following immunological parameters were studied: peripheral blood T lymphocyte subpopulations (CD3, CD4, CD8) and secretion of the Th1-type cytokines (IL-2, IFN-gamma), Th2-type cytokines (IL-6) and TGF-beta 1 by phytohaemaglutinin-stimulated peripheral blood lymphocytes. Evaluation of immunity parameters were performed before and 2 weeks after immunotherapy. RESULTS: It was found that paternal lymphocytes immunization significantly increase the percentage of CD4 T lymphocytes (37.11 +/- 7.65 vs. 41.38 +/- 5.57, p = 0.007). Immunotherapy also leads to a significant enhancement in Th2-type cytokines (IL-6) secretion (22,677 +/- 17,907 mg/ml vs. 44,550 +/- 15,907 mg/ml, p = 0.008) and a significant decreasing in Th1-type cytokines (IL-2) secretion (6.50 +/- 5.98 mg/ml vs. 0.00 +/- 0.00 mg/ml, p = 0.0179). CONCLUSIONS: The data of the present studies suggest that paternal lymphocytes immunization modulate of immunity in women with unexplained recurrent spontaneous abortion. Our studies indicate a shift in the balance fo cytokine profiles away from Th1-type reactivity to a Th2-type reactivity after immunotherapy.     

Study on the activity and phenotype of decidual natural killer cells in patients with unexplained habitual abortions

Habitualabortionsoccurin 1to 2 %ofthechild bearingpopulation .Chromosomalaberrationistheprincipalcauseoffetallossduringtheearlystageofgestation .Oth eretiologies ,whichincludeanatomicanomalies ,endocrinedisorders,andinfections ,havealsobeendocumemtedinpatientswithhabitualabortions .Nevertheless ,in 4 0to6 0 %ofcouples ,habitualabortionsremainunexplained ,whicharecalledUHA .Recently ,theimportanceofim munologyinUHAisrecognized .Immunecellsareabun dantinhumandeciduaandarecapableofrespondingto…     

Enhancement of peripheral blood CD56(dim) cell and NK cell cytotoxicity in women with recurrent spontaneous abortion or in vitro fertilization failure

Recent studies support the concept that NK cells play an important role in the success or failure of embryo implantation. Recurrent spontaneous abortion (RSA) is the most common complication of pregnancy. Some couples suffer from infertility of unknown cause. In vitro fertilization (IVF) is one of the useful treatment methods used for treatment of this type for infertility with variable outcomes. The aim of this study was to compare the percentage of peripheral blood CD56(+) (CD56(dim) and CD56(bright)) cells and the level of NK cell cytotoxicity in patients with RSA and patients with IVF failure with those of healthy multiparous and successful IVF control women. In this case-control study peripheral blood samples from 43 patients, which included 23 women with RSA and 20 with IVF failure, plus 43 healthy control women comprising 36 normal multiparous women and 7 women with successful IVF, were collected. The percentage of peripheral blood NK cells (CD56(+)) was identified by flow cytometry, then peripheral blood mononuclear cells (PBMC) were isolated by density centrifugation (Ficol-Hypaque) and incubated with NK-sensitive K562 cells. The NK cell cytotoxicity level was determined by lactate dehydrogenase (LDH) release assay. The percentage of CD56(dim) cells and the level of peripheral blood NK cell cytotoxicity in RSA patients and women with IVF failure were significantly higher than in both the healthy multiparous and successful IVF control groups (P<0.001). The findings of the present study suggest that increases in the percentage of CD56(dim) cells and NK cytotoxicity in peripheral blood may be important contributing factors for both RSA and IVF failure.     

Expression of CD28 and cytotoxic T lymphocyte antigen 4 at the maternal-fetal interface in women with unexplained pregnancy loss.

OBJECTIVE: To investigate the expression of CD28 and CTLA-4, two costimulatory molecules involved in T-cell activation at the maternal-fetal interface in women with unexplained pregnancy loss. METHODS: A total of 57 women, 39 with unexplained spontaneous abortion (SA) and 18 with unexplained recurrent spontaneous abortion (RSA), were enrolled in the study. A high-resolution spectratyping analysis of fluorescent bands was performed to detect CD28 and CTLA-4 expression in decidual tissues. RESULTS: The mean expression of CTLA-4 mRNA was significantly higher in women with SA than in controls (P<0.05). Moreover, the expression of CTLA-4 was higher in SA patients with genotype AA and phenotype A+ (AA+AG) than in controls (P<0.01). The expression of CTLA-4 was not significantly different in patients with RSA and in controls. The expression of CD28 was significantly decreased in patients with RSA (P<0.01) and SA (P<0.05) compared with controls. The mean ratios of CTLA-4/CD28 were significantly higher in patients with RSA (P<0.01) and SA (P<0.05) than in controls. CONCLUSIONS: This study suggests that an imbalance in CTLA-4/CD28 expression or suppressed T-cell activity at the maternal-fetal interface may confer susceptibility to unexplained pregnancy loss.     

淋巴细胞免疫治疗对小鼠自然流产模型母胎界面CD80~+表达及妊娠结局的影响

目的:评价CBA/J×DBA/2小鼠配对组合作为反复自然流产模型的生殖力特点,及其与母胎交界CD80表达间的关系,并研究淋巴细胞免疫治疗(lymphocyte immunotherapy,LIT)对CD80表达水平的影响。方法:对CBA/J×DBA/2小鼠的生殖力特点进行为期120d的观察,并与生殖力正常的4种对照组进行比较。另计算15对CBA/J×DBA/2小鼠孕13d的胚胎吸收率,并用CD80-FITC和CD45-PE双色流式细胞术检测CD80细胞在母胎交界面的构成比。为了明确CD80~+细胞的身份,检测了CD3、DX5(NK细胞)和MHC-Ⅱ在CD80细胞群中的表达水平。此外,检测LIT组与未治疗组CBA/J×DBA/2小鼠胚胎吸收率和CD80细胞的阳性率。结果:CBA/J×DBA/2小鼠的流产特点是为孕10d左右的反复流产。CBA/J×DBA/2小鼠孕13d的胚胎吸收率显著高于BALB/c×DBA/2小鼠(30.8％±16.6％vs.7.7％±6.7％,P相似文献   

The influence of paternal lymphocytes immunization on percentage of peripheral blood CD16+/CD56+ cells in women with primary recurrent spontaneous abortion

OBJECTIVES: Paternal lymphocytes immunization has been proposed for several years as an efficient treatment for unexplained Recurrent Spontaneous Abortion (RSA), however precise mechanism that underline the benefits of this immunotherapy is still unclear. DESIGN: The aim was to study the influence of paternal lymphocytes immunization on percentage of peripheral blood NK cells (CD16+/CD56+) in women with primary RSA of unknown etiology. MATERIALS AND METHODS: 48 patients with history of 3-5 (mean 3.3 +/- 0.7) consecutive primary RSA of unknown etiology were selected for the study. Immunotherapy with paternal lymphocytes, isolated from 100 ml of peripheral blood, was performed twice prior conception with a 4-week interval. The percentage of NK cells (CD16+/CD56+) was estimated using standard flow-cytometric immunofluorescent techniques for whole blood with one-step monoclonal anti-CD16/CD56 antibodies. Statistical analysis was performed with Wilcoxon test and the p value less than 0.05 was accepted as statistically significant. RESULTS: It was found that paternal lymphocytes immunization significantly decreases the percentage of NK cells in peripheral blood in women with RSA (23.9 +/- 8.5 vs. 16.2 +/- 7.0; p = 0.009). CONCLUSIONS: The data of the present studies suggest that paternal lymphocytes immunization modulate immunity in women with primary unexplained RSA significantly decreasing the percentage of NK cells (CD16+/CD56+) in peripheral blood.