靶向RNA干扰TIM4表达对T细胞的影响  

RNA interferes TIM4 expression in human that affects founction of T cells

作　　者：张琦[1] 张艳霞[1] 方勇[1] 杨李辉[1] 夏荣[1] 

机构地区：[1]复旦大学附属华山医院输血科,上海200040

出　　处：《中国输血杂志》2013年第9期832-834,共3页Chinese Journal of Blood Transfusion

基　　金：上海市自然科学基金项目面上项目(11ZR1405200);上海市公共卫生重点学科建设计划(12GWZX0202)

摘　　要：目的构建靶向RNA干扰T细胞免疫球蛋白及粘蛋白域蛋白4(TIM4)基因的慢病毒重组表达载体,分析TIM4基因对T细胞的影响。方法根据Gene Bank报道的TIM4 mRNA序列(NM_178759.4)设计干扰片段,通过基因工程方法构建靶向RNA干扰TIM4基因的慢病毒载体,体外转染DCs后与T淋巴细胞共培养,检测细胞增殖情况及细胞因子分泌水平。结果成功构建基因干扰的慢病毒载体并转染DCs。MTT实验表明TIM4-DC组T细胞的增殖受到抑制;ELISA检测显示:TIM4-DC组IL-12和IFN-γ的分泌均明显高于对照组(P<0.05)。结论 RNA干扰TIM4基因在DCs表达后,可抑制T细胞增殖的作用以及T细胞向Th2细胞分化。Objective To evaluate the founction of dendritic cell after transfection of Lentivirus vectors of TIM4 genes inhibited by RNA interference.Methods Based on the mRNA of TIM4(NM_178759.4) which is reported in Gene Bank,the interference sequence were designed and ligated to Lentivirus vector pMAGic4.1,then the recombinant vector were transfected to DCs.Proliferation ability of T cells and IL-12 and IFN-γsecretion were detected after co-culture the DCs with T cells.Results The Lentivirus vectors of RNA interference TIM4 gene were constructed.Expression of TIM4 shRNA from lentivirus vector in DCs.MTT results showed the stimulating ability of T cells proliferation was inhibited after co-culture cells in the TIM4-DC group;ELISA results found IL-12 and IFN-γ expression level significantly regulated in the TIM4-DC group.Conclusion Stimulating ability of T cells proliferation and developing to Th2 cells were weakened after TIM4 expression was inhibited in DCs.

关 键 词：T细胞免疫球蛋白及粘蛋白域蛋白4 RNA干扰 树突状细胞 TH细胞 

分 类 号：R457.11[医药卫生—治疗学]