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目的:研究妊娠子宫微环境中子宫自然杀伤细胞(uNK细胞)NKG2A和NKG2D及其相应配体的表达,探讨NKG2A与NKG2D的不平衡表达在母胎免疫耐受形成中的作用。方法:选择30例孕6-9周的正常妊娠妇女,分离其新鲜蜕膜组织,除去绒毛,分离蜕膜和外周血单个核细胞,采用流式细胞仪测定NK细胞的数量及NKG2A与NKG2D的表达;采用RT-PCR技术检测滋养层组织NKG2A与NKG2D配体人类白细胞抗原-E(HLA-E)、主要组织相容性复合体-Ⅰ类分子相关蛋白A(M ICA)mRNA的表达。结果:妊娠子宫蜕膜淋巴细胞中NK细胞约占70%,流式细胞分析的结果显示,子宫自然杀伤细胞NKG2A的表达显著高于外周血NK细胞,分别为97.86%±1.75%与33.35%±10.92%(x-±s),两者差异显著(P<0.05),在滋养层细胞中检测到其配体HLA-E的表达;而与外周血相比,uNK细胞表面NKG2D的表达与之较为相近,分别为93.21%±4.52%与97.80%±1.72%,但两者仍有显著差异(P<0.05)。在滋养层组织未检测到其相应配体M ICA mRNA的表达。结论:蜕膜中的淋巴细胞主要为NK细胞,其免疫学表型与外周血NK细胞有较大的区别,妊娠期子宫自然杀伤细胞表面高表达抑制性受体NKG2A,同时滋养层组织表达相应的配体人类白细胞抗原-E,这可能是维持母胎界面免疫耐受的重要因素。
Objective: To study the expression of NKG2A and NKG2D and their corresponding ligands in uterine natural killer cells (uNK cells) in pregnant uterine microenvironment, and to explore the role of imbalanced expression of NKG2A and NKG2D in the formation of immune tolerance of maternal fetus. Methods: Thirty healthy pregnant women aged from 6 to 9 weeks were selected and their fresh decidua were isolated. The villi were removed and the decidual and peripheral blood mononuclear cells were isolated. The number of NK cells and the number of NK cells in NKG2A and NKG2D The expressions of NKG2A and NKG2D ligands on leukocyte antigen-E (HLA-E) and major histocompatibility complex-type I molecule-associated protein A (M ICA) mRNA in trophoblast tissue were detected by RT-PCR. Results: NK cells accounted for about 70% of uterine decidual lymphocytes in pregnancy, and the results of flow cytometry showed that the expression of natural killer NKG2A in uterus was significantly higher than that in peripheral NK cells (97.86% ± 1.75% and 33.35% ± 10.92% (x- ± s), the difference between the two was significant (P <0.05), and the expression of HLA-E ligand was detected in trophoblastic cells. Compared with peripheral blood, the expression of NKG2D on uNK cells was Which were 93.21% ± 4.52% and 97.80% ± 1.72% respectively, but there was still significant difference between the two groups (P <0.05). No expression of its corresponding ligand, M ICA mRNA, was detected in trophoblast tissues. Conclusion: The majority of lymphocytes in decidua are NK cells. The immunological phenotype of NK cells is significantly different from that of peripheral blood NK cells. NKG2A is highly expressed on natural killer cells in pregnancy and the expression of NKG2A in trophoblast Of ligand human leukocyte antigen-E, which may be an important factor to maintain immune tolerance of maternal-fetal interface.