SLE相关基因IFIT1免疫调节功能的初步研究  

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

作　　者：李卓成[1] 罗蓉[1] 李延武[1] 

机构地区：[1]深圳市第二人民医院检验科,广东深圳518035

出　　处：《深圳中西医结合杂志》2008年第5期265-269,共5页Shenzhen Journal of Integrated Traditional Chinese and Western Medicine

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细胞中能诱导白细胞介素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 IFIT1, a novel gene associated with the systemic lupus erythematosus （SLE）. MethOds First an expression clone of IFIT1 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 IFIT1-EGFP were selected by standard limiting dilution method. The stable transfectants of EGFP and IFIT1-EGFP were stimulated with anti-CD3 and anti-CD28. Cell surface antigen expression were detected by flow cytometry. The apoptosis and expansion of each transfectant were detected according to the manufacture＇s instruction. The culture supematants were assayed for cytokines by sandwich ELISA. Furthermore transfection of gene silence vector which could produce anti-IFIT1-specifics small RNA duplexes partially blocked the expression of overexpressed IFIT1 in Jurkat T cell. The same assays were done by stimulation with anti-CD3 and anti-CD28. Results There were no significant difference in the expression level of the various cell surface molecules, apoptosis and expansion among Jurkat, Jurkat/EGFP, and Jurkat/IFIT1-EGFP transfectants.But it was found that the Jurkat/IFIT1-EGFP could enhance the production of IL-4 and IL-10. After knocking down the expression of IFIT1 by RNAi, the levels of IL-4 and IL-10 production were partially decreased. Conclusion The results indicate the SLE related-gene IFIT1 may play an important role in the production of eytokines and may interfere in the Th1/Th2 balance.

关 键 词：系统性红斑狼疮 WIT1 细胞因子 基因干预 

分 类 号：R593.24[医药卫生—内科学]