Itk蛋白在人PBMC增殖和细胞因子产生中的作用研究  

Study on the function of Itk protein in the proliferation and cytokines production of human PBMC

作　　者：何峰[1] 姚海兰[1] 肖宗慧[1] 韩继生[1] 刘哲伟[1] 

机构地区：[1]首都儿科研究所分子免疫室,北京100020

出　　处：《现代免疫学》2010年第1期5-8,共4页Current Immunology

基　　金：国家自然科学基金资助项目(30772025);北京市自然科学基金资助项目(7093115)

摘　　要：利用RNA干扰的方法抑制人外周血单个核细胞(PBMC)中Itk基因的表达,探讨其抑制Itk后对人PBMC增殖和炎症性相关细胞因子产生的影响。设计针对Itk基因的siRNA序列,与外源表达Itk的pEGFP-C1-hItk质粒共转染至HeLa细胞,筛选出有效的干扰序列;利用电转染的方法将有抑制Itk作用的siRNA转染至人PBMC,检测Itk抑制状态,同时观察细胞增殖率及炎症性细胞因子产生水平的变化。实验表明,Itk-siRNA有效地抑制了Itk蛋白的表达;Itk受到抑制后细胞增殖率明显低于对照组(P<0.05);几种重要炎性细胞因子IL-2、IFN-γ、IL-5、IL-10和IL-4产生水平下降。实验验证了Itk蛋白在人PBMC增殖和炎性细胞因子产生方面的重要作用。To explore the role of Itk in the regulation of the cell proliferation and the production of inflammatory cytokines through inhibition of the expression of Itk gene in human peripheral blood mononuclear cell（PBMC） by RNA interference（RNAi） method.siRNAs were designed according to the sequence of Itk and cotransfected into HeLa cells with plasmid pEGFP-C1-hItk which expresses Itk.After that,effective interference sequences were selected.siRNAs which have inhibitory effects were transfected into human PBMC by electroporation method,inhibitory states were studied,cell proliferation rates and production levels of inflammatory cytokines were tested.Expression of Itk protein were effectively inhibited by Itk-siRNA,cell proliferation rate decreased significantly compared with that of control group（P0.05）,production levels of important inflammatory cytokines as IL-2,IFN-γ,IL-5,IL-10 and IL-4 also decreased comparing to control group（P0.05）.Results of this study show that Itk protein plays an important role in the proliferation of PBMC and the production of inflammatory cytokines and provided basic foundations for developing Itk protein as therapeutic targets of inflammatory diseases.

关 键 词：ITK RNAI 人PBMC 

分 类 号：R392.12[医药卫生—免疫学]