系统性红斑狼疮相关基因功能的实验研究  

Study on the Function of the Related Gene IFIT1 in Systemic Lupus Erythematosus

作　　者：罗蓉[1] 李卓成[1] 李延武[1] 

机构地区：[1]深圳市第二人民医院检验科,518035

出　　处：《放射免疫学杂志》2009年第1期58-61,共4页Journal of Radioimmanology

基　　金：深圳市科技局计划项目(200602120)

摘　　要：目的:研究系统性红斑狼疮(SLE)相关基因IFIT1的功能。方法:首先将IFIT1从穿梭表达克隆转入真核表达克隆,然后转导进入Jurkat细胞系,用有限稀释法筛选单克隆;再用CD3和CD28刺激单克隆,检测膜分子的表达、细胞凋亡、增生、细胞因子分泌等免疫生物学指标;最后应用RNAi技术,下调IFIT1的表达,检测细胞因子分泌,比较分析IFIT1的功能。结果:IFIT1的过度表达在T细胞中能诱导IL-4和IL-10的分泌增加,下调IFIT1的转录水平后能部分下调IL-4和IL-10表达,而与细胞膜分子的表达、细胞的增生、凋亡无关。结论:SLE相关新基因IFIT1在细胞因子的产生中起了重要作用,可能与Thl/Th2的失衡有关。Objective To study the function of IFTT1, a novel defined gene associated with systemic lupus erythematosus （SLE）.Methods First an expression clone of IFTT1 was generated by performing an LR recombination reaction between the entry clone from genecopoiea and a gatewayTM destination vector. Then the vector was transfected into the Jurkat T cells. Single clone cells expressing EGFP and IFTT1 - EGFP were selected by standard limiting dilution method. The stable transfectants of EGFP and IFTT1 - EGFP were stimulated with anti - CD3 and anti - CD28. Cell surface antigen expression was dectected by flow cytometry. The apoptosis and expansion of each transfectant were detected according to the manufacture＇ s instruction. The culture supernatants were assayed for cytokines by sandwich ELISA. Further transfection of gene silence vector which could produce anti - IFTT1 - specifics small RNA duplexes partially blocked the expression of overexpressod IFTT1 in Jurkat T cell. The same assays were done by stimulation with anti - CD3 and anti - CD28. Results There were no significant diference in the expression level of the various cell surface molecules, apoptosis and expansion among Jurkat, Jurkat/EGFP, and Jurkat/IFTT1 - EGFP transfectants. But it was found that the Jurkat/lFTT1 - EGFP could enhance the production of IL -4 and IL - 10. After knocking down the expression of IFTT1 by RNM, the levels of IL -4 and IL - 10 production were partially decreased. Conclusion The results indicate the SLE related -gene IFTT1 may play an important role in the production of cytokines and may interfere in the Th1/Th2 balance.

关 键 词：红斑狼疮 系统性 IFIT1 细胞因子 基因干预 

分 类 号：R593.241[医药卫生—内科学] R394[医药卫生—临床医学]