【作者】 张洋；

【导师】 曾万江；

【作者基本信息】 华中科技大学 ， 妇产科学， 2020， 硕士

【摘要】 目的:通过临床标本检测,探讨γδT细胞亚群及其表达相关细胞因子在URSA中作用及免疫学机制。方法:经过伦理审核和知情同意后,收集我院2019年5月至2020年1月间妇产科门诊就诊人群外周血标本,按病史分为:URSA妊娠组(URSA-P)、非妊娠URSA组(URSA-UP)、正常妊娠组(H-P)和正常非妊娠组(H-UP)。借助流式细胞术检测各组患者外周血γδT细胞及其亚群、PD-1表达水平和相关细胞因子,并且借助免疫组化技术检测H-P组蜕膜γδT细胞的分布。结果:(1)流式细胞术检测4个组外周血PBMCγδT细胞及该细胞上PD-1表达的结果显示:外周血PBMCγδT细胞亚群中,Vδ2 T细胞频率最高。URSA-P组的γδT细胞上PD-1表达降低。(2)流式细胞术检测4个组外周血的γδT细胞及其细胞亚群中细胞毒性效应分子表达的结果显示:URSA-UP组Vδ1 T细胞中CD107a的表达水平显著高于HUP组,URSA-P组Vδ2 T细胞中CD107a的表达水平显著高于H-UP组,H-P组Vδ2 T细胞中CD107a的表达水平显著高于H-UP组,URSA-P组Vδ1 T细胞中穿孔素的表达水平显著高于H-P组,差异均具有统计学意义(P<0.05)。URSAP组Vδ1 T细胞中的穿孔素表达显著高于H-P组,差异具有统计学意义(P<0.05)。(3)流式细胞术检测4个组外周血γδT细胞的细胞因子表达谱的结果显示:URSA-P组Vδ2 T淋巴细胞表达IFN-γ的水平显著高于H-UP组,URSA患者(URSA-P组和URSA-UP组)的Vδ2 T细胞中分泌更多的IFN-γ,差异均具有统计学意义(P<0.05)。(4)免疫组化技术检测蜕膜组织的结果显示:少量的γδT细胞浸润蜕膜组织。结论:γδT细胞PD-1表达的下调、细胞毒性效应分子及Th1型细胞因子表达的上调与不明原因复发性流产相关。PD-1表达异常可能导致细胞毒性效应分子和Th1型细胞因子的表达增加,进而参与不明原因复发性流产的发生。更多还原

【Abstract】 Objective: To compare PD-1 expression levels and cytokine profile of γδ T cells in the peripheral blood of healthy pregnant women,URSA pregnant patients,URSA nonpregnant patients and healthy non-pregnant women,and to explore their roles in the pathogenesis of recurrent abortion.Methods: The patients who came to clinic for treatment in the hospital between May 2019 and Jan 2020 were divided into 4 groups: URSA pregnancy group(URSA-P),URSA non-pregnancy group(URSA-UP),healthy pregnancy group(HP)and healthy non-pregnancy group(H-UP).The frequencies of γδ T cell subsets,and PD-1 expression levels as well as cytokine profile of each subsets in their peripheral blood were measured by Flow cytometry(FCM).The presence of γδ T cells in the decidua was measured by IHC.Result: FCM showed that Vδ2 cells consisted the majority of γδ T cells in the peripheral blood(P <0.05).PD-1 expression on γδ T cells was significantly reduced in URSA-P group(P <0.05).FCM showed that CD107 a expression on Vδ1 cells in URSA-UP group was significantly higher than in H-UP group(P <0.05).CD107 a expression on Vδ2 cells in URSA-P group was significantly higher than in H-UP group(P <0.05).CD107 a expression on Vδ2 cells in H-P group was significantly higher than in H-UP group(P <0.05).And perforin expression on Vδ1 cells in URSA-P group was significantly higher than in H-P group(P <0.05).FCM showed that IFN-γ expression on Vδ2 cells in URSA-P group was significantly higher than in H-UP group(P <0.05)..And IFN-γ expression on Vδ2 cells in URSA patients were higher than on Vδ1 cells(P <0.05).Immunohistochemical staining revealed that there were a few γδ T cells in decidual.Conclusion: Decreased PD-1 expression and elevated expression of cytotoxic effector molecules as well as Th1 cytokines are associated with the occurrence of unexplained recurrent abortion.更多还原