足月妊娠母胎界面与系统免疫中NK细胞表型和T细胞变化的研究

目的探讨足月妊娠母胎界面及系统免疫中NK、T细胞的免疫状态变化及其相互关系。方法对10例正常足月妊娠妇女,剖宫产同时采集子宫底蜕膜、外周静脉血,通过流式细胞技术,检测其NK细胞亚群、NK细胞表面受体CD69、CD94及Th1/Th2免疫状态。结果底蜕膜和外周血中各指标结果为:CD56brightCD16-含量:(18·72±17·73)%和(0·28±0·18)%;CD56+CD69+亚群:(34·98±19·79)%和(3·33±1·27)%;CD56brightCD94+含量:(15·94±13·19)%和(1·17±1·19)%;CD56+CD16-/CD56+CD16+比值:(2·30±2·25)和(0·34±0·28);CD56+CD69+/CD56+CD94+比值:(1·21±0·66)和(0·28±0·12),差异均有统计学意义(P<0·01);底蜕膜中Th1、Th2、Tc1细胞含量及Th1/Th2、Tc1/Tc2比值与外周血均无显著性差异(P>0·05);子宫底蜕膜中自然杀伤细胞(uterine natural killer cell,uNK细胞)亚群,uNK细胞表面受体CD69、CD94,Th1/Th2免疫状态与外周血均无相关性(P>0·05)。结论与维持妊娠免疫耐受关系极大的uNK细胞,在足月妊娠底蜕膜中保持相对的活化状态,同时uNK、T细胞的变化独立于外周血中的免疫状态。     

辅助性T淋巴细胞1和2比率变化与妊娠期高血压疾病发病的关系

目的探讨辅助性T淋巴细胞(Th)1、2比率变化与妊娠期高血压疾病发病的关系。方法采用流式细胞技术,分别检测12例正常未妊娠妇女(正常未孕组)、12例正常妊娠妇女(正常妊娠组)、10例妊娠期高血压患者(高血压组)、25例子痫前期患者(包括10例轻度和15例重度,子痫前期组)的外周血及蜕膜组织(正常未孕组除外)中的Th1/Th2比率。结果正常未孕组妇女外周血中的Th1/Th2比率为10.5±1.5,正常妊娠组妇女外周血及蜕膜组织中的Th1/Th2比率分别为9.5±2.9及7.6±4.6、高血压组妇女分别为12.1±3.4及13.1±5.6、子痫前期组分别为16.8±3.8及26.7±9.4。子痫前期组外周血及蜕膜组织中Th1/Th2比率均明显高于其他各组,分别比较,差异均有统计学意义(P<0.05);且子痫前期组蜕膜组织中Th1/Th2比率明显高于外周血,两者比较,差异有统计学意义(P<0.001)。高血压组妇女的Th1/Th2比率变化处于子痫前期组和正常妊娠组之间。结论妊娠期高血压疾病患者Th1/Th2比率升高,导致Th1/Th2比率平衡紊乱可能是妊娠期高血压疾病发生的重要原因。     

子痫前期患者外周血中CD4^＋CD25^＋调节性T细胞水平及其意义

目的探讨子痫前期患者外周血CD4^＋ CD25^＋调节性T淋巴细胞（CD4^＋ CD25^＋Tr）和CD4^＋CD25^high调节性T淋巴细胞（CD4^＋ CD25^high Tr）在子痫前期发病中的作用。方法2005年10月至2006年3月对安徽省立医院子痫前期患者30例和正常早、晚期妊娠及正常非妊娠组妇女各13例、22例及15例,采用流式细胞仪检测其外周血CD4^＋ CD25^＋Tr和CD4^＋ CD25^highTr占CD4＋T细胞的比率,分析其与子痫前期的关系。结果子痫前期组妇女外周血CD4^＋CD25^＋Tr和CD4^＋ CD25^highTr表达率分别为（6.64±1.10）%和（0.53±0.17）%,明显低于正常晚期妊娠组妇女的（12.26±1.32）%和（1.18±0.05）%,两组比较,差异有统计学意义（P〈0.01）。正常非妊娠组妇女外周血CD4^＋ CD25^＋Tr表达率（10.8±1.05）%和正常早期妊娠组CD4^＋ CD25^＋Tr表达率（11.21±1.40）%比较,差异无统计学意义,但均低于正常晚期妊娠组（P〈0.05）。结论CD4^＋ CD25^＋Tr和CD4^＋ CD25^highTr可能与子痫前期的发病机制有关。     

子痫前期患者滋养细胞浸润相关基因mRNA及蛋白表达的研究

目的研究子痫前期患者胎盘滋养细胞基质金属蛋白酶(MMP)9、2及其组织抑制物(TIMP)1、2,肿瘤转移抑制基因KiSS-1 mRNA、蛋白的表达变化及其与子痫前期发病的关系。方法采用RT-PCR、免疫印迹(western blot)法对30例正常足月妊娠妇女(正常妊娠组)和10例妊娠期高血压患者(高血压组)及27例子痫前期(子痫前期组)患者胎盘滋养细胞中MMP-9、MMP-2、KiSS-1、TIMP-1和TIMP-2基因mRNA及蛋白表达水平[均以相对吸光度(A)表示]进行检测;应用明胶酶谱分析法,检测3组妇女胎盘孵育液MMP-9、MMP-2活性。结果(1)胎盘滋养细胞浸润相关基因mRNA表达水平:子痫前期组MMP-9、MMP-2 mRNA表达水平分别为0．39±0．05和0．71±0．16,均明显低于正常妊娠组的0．78±0．11和1．63±0．31,两组分别比较,差异有统计学意义(P均<0．05)。高血压组MMP-9 mRNA的表达水平明显高于重度子痫前期患者,差异有统计学意义(P<0．05)。子痫前期组胎盘滋养细胞KiSS-1 mRNA和TIMP-1 mRNA的表达水平分别为1．97±0．21和1．11±0．18,均明显高于正常妊娠组的0．69±0．27和0．65±0．19,差异均有统计学意义(P<0．05);高血压组胎盘滋养细胞KiSS-1 mRNA表达水平低于子痫前期组,但与正常妊娠组比较,差异无统计学意义(P>0．05)。重度子痫前期患者胎盘滋养细胞TIMP-2 mRNA表达水平明显高于正常妊娠组,差异均有统计学意义(P<0．05)。(2)胎盘滋养细胞浸润相关基因的蛋白表达水平:子痫前期组MMP-9、MMP-2基因的蛋白表达水平分别为1．07±0．35和0．74±0．23,均明显低于正常妊娠组的2．43±0．92和1．48±0．78,差异均有统计学意义(P<0．05)。子痫前期组胎盘KISS-1和TIMP-1蛋白表达水平分别为2．46±0．39和1．51±0．40,均明显高于正常妊娠组的0．91±0．35和0．93±0．56,差异均有统计学意义(P<0．05)。子痫前期组胎盘滋养细胞TIMP-2蛋白表达水平与正常妊娠组及高血压组比较,差异均无统计学意义(P>0．05)。(3)胎盘MMP-9和MMP-2酶比活性:子痫前期组分别为(2．67±0．53)和(1．13±0．28)灰度·g-1·L-1,均明显低于正常妊娠组的(8．44±3．70)和(3．87±1．43)灰度·g-1·L-1,差异均有统计学意义(P<0．05)。结论子痫前期患者胎盘滋养细胞促进浸润基因。MMP-9、MMP-2表达降低和抑制浸润基因KiSS-1和TIMP-1表达升高,可能在子痫前期胎盘缺血缺氧中起重要作用。     

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.     

血浆中可溶性人类白细胞抗原-G对子(癎)前期的早期预测价值

目的 探讨血浆中可溶性人类白细胞抗原-G（soluble HLA-G，sHLA-G）对子痫前期的早期预测价值。方法 采用前瞻性研究方法，在妊娠早期收集120例最初正常妊娠妇女的血浆样本，同时对其妊娠中期及晚期的血浆样本进行纵向收集，并且随访其妊娠结局。运用双抗体夹心酶联免疫吸附（ELISA）法检测血浆sHLA-G水平。结果 120例在早孕期最初正常的妇女中有19例随后发生子痫前期，101例妊娠结局正常。随妊娠进展，子痫前期组与正常妊娠组的血浆sHLA-G水平均有下降趋势，但无统计学意义（P〉0．05）。妊娠早期、中期、晚期子痫前期组血浆sHLA-G水平[分别为（1．25±0．02）mg／ml、（1．11±0．05）mg／ml、（0．98±0．03）mg／ml]均显著低于正常妊娠组[分别为（1．95±0．03）mg／ml、（1．90±0．02）mg／ml、（1．86±0．05）mg／ml，P均〈0．05]。早孕期血浆sHLA—G预测子痫前期的ROC曲线下面积为0．875。以sHLA-G等于1．57mg／ml为切点预测子痫前期的敏感性、特异性、阳性预测值、阴性预测值及Kappa指数分别为95％、84％、70％、97％、0．645。结论 妊娠期血浆sHLA-G水平的降低与子痫前期的发生有密切关系。妊娠早期检测血浆sHLA-G水平将有助于早期预测子痫前期。     

妊娠期高血压疾病患者胎盘组织中溶血磷脂酸受体蛋白Edg4、7的表达及其意义

目的 探讨胎盘组织中溶血磷脂酸受体蛋白Edg4、7的表达与妊娠期高血压疾病发生的关系及作用机制。方法 采用免疫组化链霉菌抗生物素蛋白-过氧化物酶（SP）法,检测20例正常晚期妊娠妇女（正常晚孕组）、20例妊娠期高血压患者（妊娠期高血压组）、20例轻度子痫前期患者（轻度子痫前期组）、30例重度子痫前期患者（重度子痫前期组）的胎盘组织中溶血磷脂酸受体蛋白Edg4与Edg7的表达。结果（1）表达部位：Edg4与Edg7主要表达于胎盘绒毛滋养细胞及蜕膜细胞的细胞质和细胞膜。（2）Edg4和Edg7在胎盘绒毛滋养细胞中的表达阳性率：正常晚孕组分别为25％和20％,妊娠期高血压组分别为60％和40％,轻度子痫前期组分别为80％和65％,重度子痫前期组分别为83．3％和86,7％。轻、重度子痫前期组阳性率明显高于正常晚孕组,两组分别比较,差异有统计学意义（P〈0,05）;妊娠期高血压组与正常晚孕妇组比较,差异无统计学意义（P〉0．05）。（3）Edg4和Edg7在胎盘蜕膜细胞中的表达阳性率：正常晚孕组分别为20％和25％,妊娠期高血压组分别为55％和50％,轻度子痫前期组分别为70％和55％,重度子痫前期组分别为83．3％和73．3％。轻度子痫前期组及重度子痫前期组阳性表达率明显高于正常晚孕组,两组分别比较,差异有统计学意义（P〈0．05）;妊娠期高血压组与正常晚孕组比较,差异无统计学意义（P〉0．05）。结论 妊娠期高血压疾病患者胎盘组织中溶血磷脂酸受体蛋白Edg4、7呈高表达,提示溶血磷脂酸与胎盘组织中的特异性受体Edg4、7结合,并参与了妊娠期高血压疾病的发生。     

子痫前期患者胎盘组织中转化生长因子β1、血管细胞黏附分子1及E选择素表达的研究

目的探讨子痫前期患者胎盘组织中转化生长因子β1(TGF-β1)、血管细胞黏附分子1(VCAM-1)和E选择素(E-selectin)的表达变化及其意义。方法用免疫组化链霉菌抗生物素蛋白-过氧化物酶连接(SP)法,对20例正常孕妇(对照组)和40例子痫前期患者(子痫前期组,其中轻度16例、重度24例)的胎盘组织进行TGF-β1、VCAM-1和E-selectin定位,并用计算机图像分析系统进行定量分析比较。结果(1)1FGF-β1在胎盘绒毛合体滋养细胞的表达,子痫前期组为70．7±0．5,对照组为70．3±0．6,两组比较,差异有统计学意义(P<0．05)。VCAM-1的表达,子痫前期组为82．5±0．5,对照组为82．8±0．3;E-selectin的表达,子痫前期组为53．5±0．5,对照组为53．8±0．4;两组分别比较,差异均有统计学意义(P<0．05)。TGF-β1、VCAM-1、E-selectin在轻度子痫前期患者胎盘组织合体滋养细胞中的表达分别为70．6±0．6、82．4±0．6、53．4±0．5,在重度子痫前期患者胎盘组织中的表达分别为70．8±0．4、82．6±0．5、53．6±0．5,轻度与重度子痫前期患者TGF-β1、VCAM-1、E- selectin表达水平比较,差异均无统计学意义(P>0．05)。(2)E-selectin在胎盘绒毛毛细血管内皮细胞的表达,子痫前期组为63．0±0．5,对照组为62．6±0．4,两组比较,差异有统计学意义(P<0．05);轻度子痫前期患者为63．2±0．4、重度子痫前期为62．9±0．5,二者比较,差异无统计学意义(P> 0．05)。结论TGF-β1、VCAM-1和E-selectin在胎盘组织中的表达变化,在子痫前期发病中有重要作用。     

子痫前期患者辅助性T淋巴细胞及协同刺激分子CD28/CTLA-4的表达

目的:探讨辅助性T淋巴细胞及协同刺激分子CD28/CTLA-4在子痫前期发病中的作用。方法:采集22例正常妊娠妇女(正常妊娠组)及45例子痫前期患者(子痫前期组,包括22例轻度和23例重度)外周血,用流式细胞技术分别检测外周血Th1、Th2及Th1/Th2比率及CD28和CTLA-4在CD4+T细胞的表达。结果:子痫前期组Th1、Th1/Th2、CTLA-4均高于正常妊娠组,且重度高于轻度(P0.05);子痫前期组Th2、CD28/CTLA-4低于正常妊娠组,且重度低于轻度(P0.05)。CD28与Th1呈负相关,r=-0.295;CTLA-4与Th1呈正相关,r=0.551,与Th2呈负相关,r=-0.363;CD28/CTLA-4与Th1/Th2呈负相关,r=-0.707。结论:子痫前期患者外周血协同刺激分子CD28/CTLA-4表达异常,高表达的CTLA-4促进CD4+T淋巴细胞过度活化并向Th1亚群分化,导致Th1/Th2比率失衡,这可能是子痫前期发生的重要原因之一。     

重度妊高征中Bax、Bcl-2表达情况的研究

目的研究胎盘和蜕膜中促(抑)凋亡基因Bax／Bcl-2表达及其在重度妊高征发生、发展中的作用。方法对40例正常晚孕和40例重度妊高征病例的胎盘和蜕膜组织进行分析用免疫组织化学方法(S-P法)检则促(抑)凋亡基因Bax／Bcl-2表达水平。结果正常晚孕组胎盘和蜕膜Bax分别是28．95％±9．67％,42．21％±4．70％;Bcl-2分别是22．91％±7．69％,43．15％±9．18％;Bax／Bcl-2分别是1．20,0．98;重度妊高征组胎盘和蜕膜Bax分别是42．46％±11．70％,60．04％±13．79％,比正常晚孕显著增高(P＜0．01);Bcl-2分别是22．34％±7．78％,33．91±6．46％,比正常晚孕增高(P＜0．05);Bax／Bcl-2分别是2．90,2．38,正常晚孕组中Bax/Bcl-2表达平衡,而重度妊高征组中这种平衡关系被打破。结论正常晚孕组和重度妊高征组中都有一定量的Bax、Bcl-2表达;重度妊高征组中,这种表达增强;Bax／Bcl-2表达间的平衡决定胎盘蜕膜的命运,进一步影响妊娠的结局。     

Relation between plasma endothelin 1 levels and T helper 1: T helper 2 cell immunity in women with preeclampsia.

The aim of this study was to investigate the relationship between plasma endothelin 1 (ET-1) levels and T helper (Th)-1:Th2 cell immunity in women with preeclampsia. The percentage of Th1 and Th2 cells and the Th1:Th2 cell ratios in peripheral blood from 11 normal pregnant women and 11 patients with preeclampsia at 29-34 weeks of gestation were calculated using flow cytometry. The plasma ET-1 level was also determined using a modified radioimmunoassay. The plasma ET-1 concentrations and the Th1:Th2 cell ratios in normal pregnancies were significantly lower than those in patients with preeclampsia. Negative correlations were found between plasma ET-1 levels and Th2 cells in both the preeclamptic pregnancy groups and in the normal pregnant women. Our results indicate that elevated ET-1 levels are associated with a Th1:Th2 imbalance in preeclampsia.     

Decidual leucocyte populations in early to late gestation normal human pregnancy

Most research on human decidual leucocytes to date has focused on the predominant CD56+ uterine natural killer (uNK) cell population in early pregnancy. Few reports have documented decidual leucocyte populations after 13 weeks gestation and in late pregnancy. Placental bed (decidua basalis) and non-placental bed (decidua parietalis) biopsies from normal pregnancies were taken from women undergoing termination of pregnancy in the 1st and 2nd trimesters and following Caesarean section in the 3rd trimester. Immunohistochemistry was used to quantify the numbers of decidual cells expressing CD56, CD3, CD8, CD94, NKG2A and CD14 and double labelled CD161+CD3+ NKT-like cells. Although a significant reduction in CD56+ uNK cells was found in 3rd trimester samples compared with 1st and 2nd trimester decidua, a substantial residual CD56+ leucocyte population was identified in 3rd trimester decidua. Expression of the KIR CD94/NKG2A mirrored that of CD56 at all gestational ages, providing an explanation for the absence of cytotoxic responses at the fetal–maternal interface. There was no difference in leucocyte populations between decidua basalis and decidua parietalis. Double immunohistochemical labelling revealed small numbers of decidual CD3+CD56+ and CD8+CD56+ cells, which decreased in number at term, and CD161+CD3+ cells, which increased in number at term. No differences in leucocyte populations were detected between decidua parietalis and decidua basalis. In contrast to previous reports, a substantial residual CD56+ cell population was demonstrated in 3rd trimester decidua. Decidual cytotoxic T-lymphocytes did not alter in number during gestation, while in contrast CD14+ macrophages decreased at term, representing the smallest decidual population assessed.     

Lymphocyte subset distribution and cytokine secretion in third trimester decidua in normal pregnancy and preeclampsia

BACKGROUND: Excessive Th1 activity in peripheral blood plays a probable role in the pathogenesis of preeclampsia. The aim of the study was to investigate whether disturbed local immune reactions are also present in decidua. METHODS: Flow cytometric analysis of CD3, CD19, CD56/CD16, CD4, CD8, CD4/CD29, CD4/CD45RA, CD4/CD45RO, CD8/CD28, CD3/CD69 lymphocyte subsets isolated from third trimester decidua of pregnants with preeclampsia (n=21) and pregnant controls (n=11) subjected to elective caesarean sections. Spontaneous and phytohemaglutynine stimulated "in vitro" secretion of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta by decidual lymphocytes was studied by ELISA. For the statistical significance of differences between the groups the U Mann-Whitney test was performed (confidence interval P<0.05). RESULTS: Preeclamptic patients were characterized with an increased percentage of the CD3-/CD56+CD16+, CD8+/CD28+ and decreased percentage of CD3+, CD19+, CD4+/CD45RA+ lymphocytes. The profile of secreted cytokines shifts in favor of Th1 activity (extremely high IFN-gamma and low IL-6 and IL-10 secretion). Decidual IL-12 secretion in preeclamptic patients is decreased compared to controls. CONCLUSION: Changes in NK and T lymphocyte subsets followed with Th1 cytokine IFN-gamma over-activity, could affect local immunoregulatory mechanisms in third trimester decidua of preeclamptic patients.     

Uterine natural killer cells dysregulation in idiopathic human preterm birth: a pilot study

Objective: To compare between uterine natural killer (uNK) cells in the placental samples of preterm birth and term labor.

Study design: Two-arm case–control study. This study included 60 participants divided into two groups. The first group included 30 cases of idiopathic spontaneous preterm labor and the other group included 30 women who delivered by a spontaneous term vaginal delivery and with no history of previous preterm labor.

Result(s): There were no CD16^? CD56^bright uNK cells in either groups; CD16⁺ CD56^dim uNK cells were present in only 1 case out of 30 term delivery placentae (3.3%), whereas they were found in 21 cases out of 30 (70%) preterm placental samples with a significant statistical difference (p?+CD56^dim uNK cells were found to be invading both the villi and the decidua in 11 cases (70%), whereas those cells were found invading only the villi in 10 cases (33.3%).

Conclusion: CD16⁺CD56^dim cells are expressed in both the decidua and the villi of patients with idiopathic preterm labor suggesting an association between uNK cells dysregulation and idiopathic human preterm labor.     

孕酮和IL-15对人早孕蜕膜子宫自然杀伤(uNK)细胞促血管生成因子的调节

目的:探讨孕酮和IL-15对蜕膜子宫自然杀伤细胞(uNK)在早孕蜕膜血管生成/重塑过程中的作用。方法:免疫磁珠(MACS)分离及纯化蜕膜uNK细胞后进行体外培养,分别用不同浓度孕酮或IL-15干预72h后获取培养上清液和细胞。采用RT-PCR和ELISA检测蜕膜uNK细胞的VEGF-A、VEGF-C、Ang2的mRNA转录和蛋白表达。结果:蜕膜uNK细胞可表达多种与血管形成有关的生物活性分子,与未干预的空白对照组相比,IL-15促进uNK细胞表达VEGF-A、VEGF-C(P<0.05),且呈剂量依赖关系,但对Ang2无影响;而孕酮对uNK细胞表达上述因子均无显著影响(P>0.05)。结论:妊娠早期蜕膜uNK细胞通过表达多种促血管生成因子,在早孕蜕膜血管生成/重塑中起着重要的作用,IL-15对此有直接促进作用。     

The Urokinase Plasminogen Activator (uPA) System in Uterine Natural Killer Cells in the Placental Bed During Early Pregnancy

The urokinase plasminogen activator (uPA) system plays pivotal roles in cell invasion, adhesion and migration. Roles for uterine natural killer (uNK) cells in regulating extravillous trophoblast (EVT) invasion and spiral artery remodeling have been proposed. Placental bed biopsies from early pregnancy were obtained from three gestational age groups (8–10, 12–14 and 15–20 weeks). Total caseinase activity in the placental bed was studied using casein in situ zymography. Localisation of uPA, uPA receptor (uPAR), plasminogen activator inhibitor (PAI)-1 and -2 in the placental bed was investigated by immunohistochemistry. CD56⁺ uNK cells were separated from collagenase-digested decidual cells using an immunomagnetic technique, and uPA activity was measured in isolated cell culture supernatants by casein/plasminogen gel zymography (8–10 and 12–14 weeks' gestation, n = 10 each group). uPAR in cell lysates and PAI-1 and -2 secretion in supernatants were measured by Western blotting. Caseinase activity was stronger in decidua than myometrium as shown by in situ zymography. uPA localised strongly to uNK cells, especially at 8–10 weeks. Moderate uPAR localisation on uNK cells also observed. There was very weak immunostaining of uNK cells for PAI-1 and PAI-2. In casein gel zymography, uPA activity was similar in uNK cell culture supernatant compared with total unseparated decidual cells. uPAR in uNK cell lysates was significantly stronger than in total decidual cell lysates. PAI-1 and PAI-2 were not detected in uNK cell culture supernatants by Western blot analysis. These results suggest that uNK cells may regulate EVT invasion and spiral artery remodeling via the uPA system.     

主动免疫治疗对不明原因习惯性流产患者辅助T细胞因子水平的影响

目的　探讨主动免疫治疗对不明原因习惯性流产 (UHA)患者辅助T细胞 (Th) 1 /Th2型细胞因子水平的影响。方法　采用酶联免疫吸附法 ,检测 30例半年内接受过淋巴细胞主动免疫治疗的UHA患者 (治疗组 ) ,及 2 5例未经治疗的UHA患者 (未治疗组 ) ,外周血单个核细胞 (PBMC)经滋养细胞抗原刺激产生的Th1型细胞因子白细胞介素 (IL) 2、γ干扰素 (IFN γ)及Th2型细胞因子产生IL 4、IL 1 0水平。并选取 1 5例正常非妊娠妇女作为对照 (对照组 )。结果　 (1 )在最佳诱导时间内 ,治疗组IL 2、IFN γ的水平分别为 (1 0 8± 37)ng/L、(1 1 0± 52 )ng/L ,明显低于未治疗组的 (2 2 3± 85)ng/L、(32 6±92 )ng/L(P值均 <0 .0 5) ;IL 4、IL 1 0水平分别为 (50± 1 1 )ng/L、(1 4 0± 37)ng/L ,明显高于未治疗组的(2 3± 1 1 )ng/L、(52± 2 8)ng/L(P值均 <0 .0 5)。未治疗组IL 2、IFN γ水平明显高于对照组的 (92± 32 )ng/L、(1 0 2± 35)ng/L(P值均 <0 .0 5) ;IL 4、IL 1 0水平低于对照组的 (62± 2 1 )ng/L、(1 50± 42 )ng/L(P值均 <0 .0 5)。治疗组与对照组各细胞因子水平比较 ,差异均无显著性 (P值均 >0 .0 5)。 (2 )治疗组30例患者治疗后半年内妊娠 2 6例 ,其中 8例自然流产 ,IL 2、IFN γ水平明显高于 1 8例妊娠     

A study on the density and distribution of uterine Natural Killer cells at mid pregnancy in mice genetically-ablated for CCR2, CCR 5 and the CCR5 receptor ligand, MIP-1 alpha

Several chemoattractants mediate Natural Killer (NK) cell migration. The CC-chemokines, monocyte inflammatory protein (MIP)-1 alpha, regulated upon activation, normal T cell expressed and secreted (RANTES) and macrophage chemotactic protein (MCP)-1 are the most potent. Peripheral NK cells express the CC-chemokine receptor CCR2, for MCP-1 and CCR5, for MIP-1 alpha and RANTES. These chemokines are detected in the uterus during the estrous cycle and become elevated during pregnancy. To assess the roles of CCR2, CCR5 and MIP-1 alpha in NK cell migration to the uterus and localization within implantation sites, histological analysis was conducted on implantation sites from mice genetically-ablated for CCR2, CCR5, MIP-1 alpha or CCR2 and MIP-1 alpha. Uterine NK (uNK) cell densities in both the decidua basalis and mesometrial lymphoid aggregate of pregnancy (MLAp) of all mutant strains matched wildtype controls. Ratios of vascular: non-vascular uNK cell position were identical in mutants and controls. In the decidua basalis, 25-35% and in the MLAp, 15-20% of uNK cells were perivascular. Intravascular uNK cells were observed in the decidua basalis but not in the MLAp and were more numerous at gestation day 10 than 12. Two measures of uNK cell activation, cell diameter and cytoplasmic granule number, were similar in the mutants and controls. Thus, migration, distribution and activation of NK cells within the pregnant uterus are independent of CCR2, CCR5 and MIP-1 alpha.     

Uterine NK cell development, migration and function

Uterine natural killer (uNK) cells represent the predominant lymphocytes in the uterus during early pregnancy and in the secretory phase of the menstrual cycle. They are CD56(high)CD16(-) and have low cytotoxicity, but constitutively secrete a number of cytokines, chemokines and angiogenic molecules. uNK cells differ from CD56(high) blood NK cells in several ways, including the killer cell immunoglobulin-like receptor repertoire and expression of some genes induced by hormone environment. uNK cells may arise by in-utero proliferation and differentiation of NK cell progenitors under the control of the sex steroid hormones and/or cytokines, such as interleukin-15, and/or be recruited from CD56(+) blood NK cells that would undergo tissue-specific differentiation in the uterine microenvironment. There is evidence showing that uNK cells display a different pattern of chemokine receptors and adhesion molecules, thus leading to a different migratory response. It has not yet been fully defined which uNK cell function(s) are critical for successful pregnancy. The close encirclement of spiral arteries by NK cells, together with their ability to produce angiogenic factors, suggests that they might influence mucosal vascularization. Their proximity to the extravillous trophoblast supports the idea that uNK cells could recognize these cells as fetal, and regulate their invasion during placentation.