汉滩病毒感染血管内皮细胞引起TLR4表达上调

Hantaan virus infection of vascular endothelial cell leads to up-regulation of Toll-like 4 expression

目的探讨汉滩病毒感染血管内皮细胞后,Toll样受体(TLRs)分子的表达变化,为抗感染免疫和致病机制研究提供重要资料。方法分别用LPS,CL097,PolyI:C以及汉滩病毒76-118刺激血管内皮细胞,6h后提取总RNA,进行反转录获得cDNA,再分别用各自的引物进行PCR反应,产物用1%琼脂糖凝胶电泳分析,并用间接免疫荧光测定汉滩病毒感染血管内皮细胞后TLR4的表达。结果汉滩病毒刺激感染后,血管内皮细胞TLR2、TLR4转录水平增高,TLR3转录水平降低,细胞膜表面的TLR4表达上调。结论汉滩病毒76-118感染血管内皮细胞后,可以引起TLRs分子转录水平发生改变,其中TLR4表达上调,固有免疫参与了汉滩病毒致病过程。

Objective To investigate the transcription and expression of Toll-like receptors of EVC-304 cells infected by Hantavirus. Methods LPS,CL097,Poly I:C,and Hantavirus 76-118 were used to affect EVC-304 cells. The total RNA of cells was obtained at 6 h postinfection. The transcription of TLR2,TLR3,TLR4,TLR7,and TLR9 was detected by RT-PCR. The RT-PCR products were identified by 1% agarose electrophoresis. The expression of TLR4 in EVC-304 was observed by indirect immunofluorescence assay. Results Hantavirus affection up-regulated TLR2 and TLR4 but down-regulated TLR3 of EVC-304 cells at transcription level. The expression of TLR4 on the surface of EVC-304 was up-regulated by the affection. Conclusion The transcription of TLRs of ECV-304 has a change after the cells were infected by Hantavirus and the expression of TLR4 was up-regulated,which indicates that innate immune response may participate in the pathopoiesis of Hantavirus.