文章摘要
周小斌,费小蔷,叶 军,郭鹏飞,周成林.T1DM患者外周血记忆性T细胞表面PD⁃1表达水平与其疾病发生发展的初步研究[J].南京医科大学学报,2018,(4):476~481
T1DM患者外周血记忆性T细胞表面PD⁃1表达水平与其疾病发生发展的初步研究
Preliminary research on the expression levels of PD-1 on memory T cells in the peripheral blood from T1DM patients with the development and progression of disease
投稿时间:2017-05-16  
DOI:10.7655/NYDXBNS20180410
中文关键词: 1型糖尿病  2型糖尿病  PD⁃1  记忆性T细胞
英文关键词: type 1 diabetes mellitus  type 2 diabetes mellitus  PD⁃1  memory T cells
基金项目:泰州市人民医院院级课题(ZL201614)
作者单位
周小斌 泰州市人民医院检验科江苏 泰州 225300 
费小蔷 泰州市人民医院内分泌科江苏 泰州 225300 
叶 军 泰州市人民医院中心实验室江苏 泰州 225300 
郭鹏飞 上海市第十人民医院检验科上海 200070 
周成林 泰州市人民医院检验科江苏 泰州 225300 
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中文摘要:
      目的:通过检测1型糖尿病(type 1 diabetes mellitus,T1DM)、2型糖尿病(type 2 diabetes mellitus,T2DM)患者以及健康对照外周血T细胞亚群免疫负调控分子程序性死亡蛋白1(programmed cell death protein 1,PD?1)mRNA相对表达水平和记忆性T细胞(memory T cells,Tm)表面PD?1表达情况,从分子生物学、细胞免疫学等多种角度研究记忆性T细胞PD?1表达异常与以胰岛β细胞破坏为主要机制的T1DM发生发展的相关性。方法:分离T1DM、T2DM患者以及健康对照组外周血单个核细胞(peripheral blood mononuclear cells,PBMCs);从PBMCs中分离出CD4+T细胞和CD8+T细胞,使用荧光定量PCR分别检测其细胞内PD?1 mRNA相对表达水平;将PBMCs分别标记荧光抗体CD4?FITC、CD45RO?PE、CCR7?APC、CD8?FITC、PD?1?PerCp,应用流式细胞术分别检测CD4+CD45RO+CCR7+Tcm细胞、CD4+CD45RO+CCR7-Tem细胞、CD8+CD45RO+CCR7+Tcm细胞、CD8+CD45RO+CCR7-Tem细胞表面免疫负调控分子PD?1的表达水平。结果:①T1DM组患者外周血中CD4+T细胞内PD?1 mRNA相对表达水平低于T2DM组和正常对照组,差异有统计学意义;②T1DM组患者外周血中CD8+T细胞内PD?1 mRNA相对表达水平未见明显异常;③T1DM组患者外周血中CD4+CD45RO+CCR7-Tem细胞亚群与CD4+CD45RO+CCR7+Tcm细胞亚群PD?1表达水平均显著低于正常对照组和T2DM组患者,差异有统计学意义;④T1DM组患者外周血中CD8+CD45RO+CCR7-Tem细胞亚群与CD8+CD45RO+CCR7+Tcm细胞亚群PD?1表达水平均无明显异常。结论:胰岛β细胞可通过细胞表面PD?L1与CD4+Tm细胞表面PD?1结合从而负调节后者细胞免疫作用,因此当CD4+Tm细胞PD?1表达异常而失去负调控作用时,细胞效应则会进一步增强,最终可能通过破坏胰岛β细胞而导致T1DM的发生发展。
英文摘要:
      Objective:Through the detection of the relative abundance of immune negative control molecules programmed cell death protein 1(PD?1) mRNA in T?cell subsets and the PD?1 expression on memory T cells from peripheral blood in type 1 diabetes mellitus(T1DM) patients,type 2 diabetes mellitus(T2DM) patients and healthy control separately,to further study the relationship of abnormal expression of PD?1 on memory T cells with the development and progression of T1DM which is characterized by pancreatic β?cell destruction. Methods:Peripheral blood mononuclear cells(PBMCs)were isolated from health control,T1DM patients,and T2DM patients separately;PBMCs were further sorted into CD4+T cells and CD8+T cells,then the relative abundance of PD?1 mRNA was analyzed by fluorescence quantitative PCR;PBMCs were marked by fluorescent antibody CD4?FITC,CD45RO?PE,CCR7?APC,CD8?FITC and PD?1?PerCp separately,then analyzed the expression of PD?1 on CD4+CD45RO+CCR7+Tcm cells,CD4+CD45RO+CCR7-Tem cells,CD8+CD45RO+CCR7+Tcm cells and CD8+CD45RO+CCR7-Tem cells by flow cytometry(FCM). Results:①The relative abundance of PD?1 mRNA in CD4+T cells from the peripheral blood of the T1DM group was significantly lower than that of the T2DM group and the healthy control group;②There was no abnormality for the relative abundance of PD?1 mRNA in CD8+T cells from the peripheral blood of the T1DM group,compared with the T2DM and healthy control groups;③The expression levels of PD?1 on CD4+CD45RO+CCR7-Tem cells and CD4+CD45RO+CCR7+Tcm cells in the peripheral blood from T1DM patients were significantly lower than those of the healthy control and T2DM group;④There was no significant difference for the expression of PD?1 on CD8+CD45RO+CCR7-Tem cells and CD8+CD45RO+CCR7+Tcm cells between the T1DM group,the T2DM group and the healthy control group. Conclusion:For PD?L1 which is expressed on islet β cells can combine with PD?1 on CD4+Tm cells to negatively regulate cell immunity,so when the expression of PD?1 on CD4+Tm cells is abnormal,cell effect will be further enhanced for the loss of negative regulation,so it may eventually lead to the development of T1DM by destroying islet β cells.
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