Clinical reserch for diagnosis of immunological habitual abortion

【导读】自然流产是妊娠最常见的并发症。据估计，有2%到5%的夫妇因为反复的自然流产而没有孩子。(1)尽管遗传、解剖、激素因素和自身免疫性疾病都与之有关，但仍有相当大比例的病例无法解释。免疫系统的原因已被认为是许多这样受影响的夫妇。由于母乳妊娠与母体同种异体反应的诱导和适应有关，不适当的同种异体反应被认为是当前许多夫妇妊娠失败的原因。因此，夫妻之间HLA抗原的共享被认为是由于不适当的同种异体反应导致习惯性流产的免疫学指标。然而，最近，这一指标的可靠性受到了质疑。(2)因此，免疫学参数可用于诊断免疫性流产和选择最可行的免疫形式，必须尽快建立更多的治疗方法。在过去的两年中，重点关注有连续两次原因不明的妊娠早期流产史且未活产的患者（研究组）的免疫学状况。我们已经调查了哪些免疫测试是有用的预测参数预后的妇女谁曾接受过治疗的后续妊娠。[结果](1)研究组68例患者NK（自然杀伤）细胞活性（以K562人髓系白血病细胞为靶点，采用铬-51释放细胞毒法测定）为39.4(]SY.+-[)12.7%，显著高于无病史的47例未怀孕健康女性（29.0=12.8%），47例未怀孕健康女性NK活性水平较高（42.0%），定义为NK活性的平均pius 1 SD；24名孕前NK活性水平升高的患者在随后的妊娠中流产率（71%）远高于44名NK活性正常的患者，后者的妊娠率仅为20%。(2)采用两种单克隆抗体双色流式细胞术鉴定外周血淋巴细胞NK亚群；研究组42例患者cd16和CD56。42例中24例维持正常妊娠（NP：非正常妊娠），42例中18例妊娠早期流产（SA：自然流产），NP中CD56(+) 16（+）百分比；6.1（]SY.+- .[）与SA相比显著降低4.1%；(SY。+ - 10.2%(4.2%)。（p<0.05）；CD56(+) 16（-）和CD56(-) 16(+)，两组间差异有统计学意义。NP组cd56阳性细胞比例为8.7(]SY.+-[)4.3%，显著低于SA组13.7%(]SY.+-[)5.3%。检测活胎和不活胎间cd56阳性细胞>=12.5%。结果灵敏度：72.4%，特异度；阳性预测值76.9%；87.5%。【讨论】清楚地揭示了复发性自然流产妊娠结局与NK细胞活性和亚群的关系。人蜕膜组织似乎在培养植入胚胎中起着重要作用，但也在防止母体免疫系统的排斥反应中起着重要作用。与子宫内膜相比，早期蜕膜组织中各种影响局部免疫的淋巴细胞和巨噬细胞数量明显增加。在这些退化细胞中，大颗粒淋巴细胞（LGL）在妊娠早期尤其丰富。每个LGL包含许多表达CD56的细胞，CD56是NK细胞的表面标记物。(3)此外，LGL的下降还揭示了NK的活性。(4)另外，由于卵泡性白细胞介素-2刺激后，卵泡性白细胞介素-2可杀死人滋养层细胞，因此可以认为卵泡性白细胞介素-2在妊娠的免疫维持中起关键作用。(5)考虑到以上结果，我们认为外周血NK细胞水平的分析可能是免疫性复发性流产的有用参数。此外。我们认为NK活性和NK子集这两个因素的组合可能是一个更准确的预测参数。少

[Introduction] Spontaneous abortion is the most complication of pregnancy. Estimates show that 2% to 5% couple me childless because of repeated spontaneous abortion. (1) Although genetic, anatomic, hormonal factors, and autoimmune disorders have been impticated, a substatial proportion of cases remain unexplained. An immunological cause has been suggested for many such affeted couples. Because manmmailan pregnancy is associated with the induction and adaptation of maternal allogeneic reactions, inappropriate allogenic responses have been suggested as a cause of tecurrent pregnancy loss in many coupies. The sharing of HLA antlgens between husband and wife has thus fur been considered as an immunological indicator of habltual abortion due to inappropriate allogenic response. However, recently, the rellability of this indicator has been thrown into doubt. (2) Therefore, immunological parameter that could be used to diagnose immunological abortion and seiect the most sultable form of immun … More otherapy must be estabilshed as soon as possible During the past two years, concentrating attentions on the immunological status of patients who had histories of two consecutive first-trimester abortions due to euexpiained cause and no live births (study group), We have investigated which immunological tests were useful predictive parameter for prognosis of subsequent pregnancy in women who had had recelved no tratment.[Resuits] (1) NK (natual killer) -cell activity (mensured by a chromium-51 release cytotoxic assay, with K562 human myeloid leukemia cells as targets) in 68 patients of the study group was 39.4(]SY.+-。[)12.7%, significant higher than that of 47 non-pregnant healthy females with no history (29.0=12.8%) With a high level of NK activity (42.0%) defined as the mean pius 1 SD of NK activity in 47 non-pregnant healthy females, the 24 patients with raised preconceptional levels of NK activity had a much higher abortion rate (71%) in the subsequent pregnancy than the 44 patlents with normal NK activity, who experlenced pregnancy loss in only 20% of pregnancles. (relative risk 3.5 ; 95% C11.8-6.5)(2) NK subsets in pripheral blood lymphpcytes were identifled by two-color flow cytometry using two monoclonal antibody ; CD 16 and CD56 in 42 patients of the study group. The 24 patients of 42 maintained the next pregnancy normally (NP : notnal prenant women ) The 18 patients of 42 aborted in the first trimester (SA : spontaneous aborters) The percentage of CD56 (+) 16 (+) in NP ; 6.1(]SY.+-。[)4.1% was significant lower compared with that in SA ; 10.2%(]SY.+-[)4.2%. (p<0.05) In other subsets ; CD56 (+) 16 (-) and CD56 (-) 16 (+), there were significant differenes between the two groups. The percentage of CD56-positive cells in NP was 8.7(]SY.+-[)4.3%, significant lower than 13.7%(]SY.+-[)5.3% in SA.Frequency of CD56-positive cells >=12.5% between viableand and nonviable pregnacy was checked out. The results were sensitivity : 72.4%, spedificity ; 76.9% positive predictive value ; 87.5%.[Discussion] The associations between pregnancy outcome of the recurrent spontaneous aborters and NK cells : activity and subsets were revealed clearly. Human decidual tissue appears to play an important role in nurturing the implanted emgryo, but also in preventing its rejection by the maternal lmmune system. Compared with the uterine endometrium, eariy decidual tissue has been shown to contain a markedly increased number of various kinds of lymphocytes and macrophages, which afect local lmmunlty. Among these decldual cells, the large granular lymphocytes (LGL) are especially abundant in the first trimester of pregnancy. Decidual LGL contain many cells expressing CD56, a surface marker of NK cells. (3) Moreover, decldual LGL have been revealed NK activity. (4) Since additionally decidual LGL could kill human trophoblasts cells after prior stimulation with interleukin-2, it is considered that decidual LGL play a pivotal role in the immunological maintenance of pregnancy. (5) Considering the above results, it is considered as possible that the analysis of NK cells in the peripheral blood leveis might be useful parameter for immunological recurrent abortion. Additionally.we supposed that the combination of two factors, NK activity and NK subset, could be a more accurate predictive parameters. Less