异基因造血干细胞移植患者外周血CD4^+T细胞中STAT3启动子区DNA甲基化水平与急性移植物抗宿主病的关系  

Relationship between the methylation status of STAT3 promoter DNA in peripheral blood CD4^+ T cells from patients after allo-HSCT and aGVHD

作　　者：徐雅靖[1] 张媛媛[1] 陈焱[1] 付斌[1] 杨晶 陈方平[1] XU Yajing ZHANG Yuanyuan CHEN Yan FU Bin YANG Jing CHEN Fangping(Department of Hematology, Xiangya Hospital, Central South University, Changsha 410008, China)

机构地区：[1]中南大学湘雅医院血液科,长沙410008

出　　处：《中南大学学报（医学版）》2017年第8期911-918,共8页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81570165)~~

摘　　要：目的:探讨异基因造血干细胞移植患者外周血CD4^+T细胞中STAT3启动子区DNA甲基化水平与急性移植物抗宿主病(acute graft versus host disease,a GVHD)的关系。方法:收集行同胞全相合异基因造血干细胞移植的40例患者的血液样本,ELISA检测各组患者血清IL-10,TGF-β1,IL-17A,IL-17F等细胞因子水平;实时定量PCR检测各组患者外周血CD4+T细胞中Treg(Foxp3,CTLA4,IL-10,TGF-β1)和Th 17(RORγt,IL-17A,IL-17F)相关基因的转录水平;实时定量PCR和Western印迹检测各组患者STAT3的表达水平;亚硫酸氢盐处理后测序(bisulfite sequencing PCR,BSP)法检测各组患者STAT3基因启动子区DNA甲基化水平。结果:与未发生a GVHD患者比较,a GVHD患者血清中IL-10及TGF-β1水平明显降低,IL-17A及IL-17F水平明显升高;a GVHD患者外周血CD4+T细胞中Foxp3,CTLA4,IL-10,TGF-β1转录水平明显降低,RORγt,IL-17A,IL-17F转录水平明显升高;a GVHD患者外周血CD4+T细胞中STAT3的表达水平明显升高,STAT3启动子区DNA甲基化水平明显降低,且STAT3表达水平与其启动子区DNA甲基化水平呈明显负相关。结论:Treg/Th17的比例失衡是异基因造血干细胞移植后患者发生a GVHD的重要因素,STAT3启动子区DNA低甲基化可能介导STAT3的过度表达,参与Treg/Th 17的比例失衡。Objective： To study the relationship between acute graft versus host disease（a GVHD） and the methylation status of the STAT3 promoter in peripheral blood CD4~＋ T cells from patients after allogeneic hematopoietic stem cell transplantation（allo-HSCT）. Methods： We collected 40 patients who underwent allo-HSCT from HLA-identical sibling donors. Serum IL-10, TGF-β1, IL-17 A and IL-17 F levels were detected by ELISA. Foxp3 cytotoxic T-lymphocyte-associated protein 4（CTLA4）, IL-10, TGF-β1, RORγt, IL-17 A and IL-17 F m RNA levels in CD4^＋ T cells were measured by real-time PCR. STAT3 expression levels were detected by real-time PCR and Western blot, and promoter DNA methylation was analyzed by bisulfite sequencing PCR（BSP）. Results： IL-10 and TGF-β1 levels were significantly down-regulated, while IL-17 A and IL-17 F levels were significantly up-regulated in patients with a GVHD compared with patients without a GVHD. Foxp3, CTLA4, IL-10, TGF-β1 m RNA levels were significantly down-regulated, while RORγt, IL-17 A, IL-17 F m RNA levels were significantly up-regulated in patients with a GVHD compared with patients without a GVHD. STAT3 expression was increased, while STAT3 promoter DNA was hypomethylated in patients with a GVHD compared with those without a GVHD. The STAT3 m RNA level was negatively correlated with STAT3 promoter DNA methylation. Conclusion： The imbalance of Treg/Th17 in CD4^＋ T cells from patients after allo-HSCT is a key factor for triggering a GVHD, and the DNA hypomethylation of STAT3 promoter could promote its expression in CD4^＋ T cells and contribute to the imbalance.

关 键 词：异基因造血干细胞移植 急性移植物抗宿主病 STAT3 DNA甲基化 

分 类 号：R457.7[医药卫生—治疗学]