NKT细胞在小鼠胚胎着床期母胎免疫耐受中作用的初步研究

发布时间：2018-03-26 13:47

本文选题：T细胞　切入点：NKT细胞　出处：《南昌大学》2010年硕士论文

【摘要】： 目的: 探究着床前后母体子宫蜕膜内T淋巴细胞的变化;通过刺激NKT细胞的增殖活化,检测其分泌的细胞因子变化,分析其在母体子宫内膜免疫微环境中的调节作用,以探明NKT细胞活化与胚胎着床的关系。方法: 1、用免疫组化法观察T细胞在围着床期小鼠子宫组织中的定位; 2、用淋巴细胞分离液分离小鼠外周血单核淋巴细胞; 3、通过腹腔注射不同浓度的抗原α-GalCer刺激NKT细胞活化,用RT-PCR法半定量检测NKT细胞活化后外周血单核细胞和子宫蜕膜组织中的IL-4,IFN-γ分泌的变化; 结果: 1、免疫组化结果显示在妊娠D1、D2、D3、D4天小鼠子宫内膜仅有少量CD3在T细胞膜上表达;妊娠D5天小鼠子宫内膜CD3表达量明显增多,并且达到高峰;妊娠D6天小鼠子宫内膜CD3表达量下降; 2、腹腔注射不同浓度(0、10.3、20.6、41.2ug/ml)的抗原α-GalCer组中,用RT-PCR技术检测不同组别外周血单核淋巴细胞和子宫蜕膜中IFN-γ的mRNA的表达。结果显示实验组各组中IFN-γ的mRNA表达明显低于空白对照组,差异有显著性(P0.05);而不同浓度药物的实验组间IFN-γ的mRNA表达量差异无显著性(P0.05); 3、腹腔注射不同浓度(0、10.3、20.6、41.2ug/ml)的抗原α-GalCer组中,用RT-PCR技术检测不同组别外周血单核淋巴细胞和子宫蜕膜中IL-4的mRNA的表达。结果显示在外周血单核淋巴细胞,实验组各组中IL-4的mRNA表达明显低于空白对照组,差异有显著性(P0.05);不同浓度药物的实验组间IL-4的mRNA表达量差异无显著性(P0.05)。而在子宫内膜,剂量为20.6ug/ml组IL-4的mRNA表达较其他组明显升高,差异有显著性(P0.05);其他各组间无显著性差异(P0.05); 4、通过分析外周血单核细胞和子宫内膜IFN-γ/IL-4的相对光密度比值,结果显示在外周血中10.3ug/ml、41.2ug/ml组IFN-γ/IL-4较空白组和20.6ug/ml组明显降低(P0.05),20.6ug/ml组与空白组相比,差异无显著性(P0.05)。而在子宫内膜,10.3ug/ml、20.6ug/ml组IFN-γ/IL-4与空白组相比,明显减少,差异有显著性(P0.05);41.2ug/ml组IFN-γ/IL-4与空白组相比,差异无显著性(P0.05)。结论: 1、CD3 T细胞在小鼠胚胎植入母体过程中增多,提示CD3 T细胞可能参与胚胎着床免疫微环境的调节; 2、着床期子宫内膜中活化的NKT细胞可能通过产生更多的IL-4使子宫蜕膜局部免疫反应朝体液免疫应答方向调节; 3、着床期外周活化的NKT细胞使外周单核细胞中IFN-γ/IL-4向Th1漂移,而子宫内膜中活化的NKT细胞使IFN-γ/IL-4的比值向Th2漂移,有利于局部子宫内膜形成免疫耐受微环境。
[Abstract]:Objective:. To explore the changes of T lymphocytes in decidua of maternal uterus before and after implantation, to detect the changes of cytokines secreted by NKT cells by stimulating their proliferation and activation, and to analyze its regulatory role in the maternal endometrium immune microenvironment. To investigate the relationship between NKT cell activation and embryo implantation. Methods:. 1. Immunohistochemical method was used to observe the localization of T cells in the uterus of mice in the surrounding bed. 2Murine peripheral blood mononuclear lymphocytes were isolated by lymphocyte isolating solution. 3. The activation of NKT cells was stimulated by intraperitoneal injection of 伪 -GalCer at different concentrations. The changes of IL-4 IFN- 纬 secretion in peripheral blood monocytes and decidua tissues after activation of NKT cells were detected by RT-PCR method. Results:. 1Immunohistochemical results showed that only a small amount of CD3 was expressed on T cell membrane in endometrium of mice on day 4 of gestation, the expression of CD3 in endometrium of mice on day 5 of gestation was significantly increased and reached its peak, and the expression of CD3 in endometrium of mice on day 6 of gestation decreased. 2. The expression of IFN- 纬 mRNA in peripheral blood mononuclear lymphocytes and decidua of different groups was detected by RT-PCR technique in the 伪 -GalCer group with different concentrations of 10.3 ~ 20.6g / ml antigen 伪 -GalCer. The results showed that the mRNA expression of IFN- 纬 in the experimental group was significantly lower than that in the blank control group. The difference was significant (P 0.05), but there was no significant difference in the expression of IFN- 纬 mRNA between the experimental groups with different concentrations of drugs. 3. The expression of IL-4 mRNA in peripheral blood mononuclear lymphocytes and decidua of different groups was detected by RT-PCR technique in the 伪 -GalCer group with different concentrations of 10.3 ~ 20.6g / ml antigen 伪 -GalCer. The results showed that the expression of IL-4 in peripheral blood mononuclear lymphocytes (PBMC) and decidua decidua was higher than that in normal controls. The expression of IL-4 mRNA in the experimental group was significantly lower than that in the blank control group (P 0.05), but there was no significant difference in the mRNA expression of IL-4 between the experimental groups with different concentrations of drugs. The mRNA expression of IL-4 in the 20.6ug/ml group was significantly higher than that in the other groups (P 0.05), but there was no significant difference among the other groups. 4. By analyzing the relative optical density ratio of IFN- 纬 / IL-4 in peripheral blood monocytes and endometrium, the results showed that IFN- 纬 / IL-4 in peripheral blood of 10.3ugr / ml 41.2ugrml group was significantly lower than that of blank group and 20.6ug/ml group. There was no significant difference between IFN- 纬 -IL-4 and control group, but there was no significant difference in IFN- 纬 -IL-4 between normal group and control group (P 0.05U / ml, P 0.05U / ml, P 0.05U / ml, P 0.05U / ml, P 0.05U / ml, P 0.05), but there was no significant difference in IFN- 纬 -IL-4 between normal group and control group (P 0.05). Conclusion:. 1CD3 T cells increased during embryo implantation, suggesting that CD3 T cells may be involved in the regulation of the immune microenvironment of embryo implantation. 2. The activated NKT cells in implantation endometrium may regulate the local immune response of decidua towards humoral immune response by producing more IL-4. 3. Peripheral activated NKT cells in implantation phase made IFN- 纬 / IL-4 drift to Th1 in peripheral monocytes, while activated NKT cells in endometrium shifted the ratio of IFN- 纬 / IL-4 to Th2, which was beneficial to the formation of immune tolerance microenvironment in local endometrium.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

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