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目的 探讨整合素 β3表达与汉滩病毒 (HTNV)感染的关系。 方法 将人整合素 β3、αv及αⅡb真核表达载体分别及共转染至CHO细胞 ,采用间接免疫荧光测定 (IFA)、流式细胞计数(FCM)检测外源基因的表达 ;然后用HTNVA9株感染稳定转染的CHO细胞 ,应用IFA和FCM检测病毒抗原。结果 整合素 β3在共转染组细胞中高效表达 ,且目的蛋白主要定位于细胞膜上 ;β3单转染组目的蛋白虽有一定量的细胞膜表达 ,但表达量低于共转染组 (P <0 0 5 ) ;αv和αⅡb单转染组目的蛋白的表达量明显低于共转染组 (P <0 0 1) ,且定位发生变化。病毒感染率共转染细胞较高 ,CHO/αvβ3和CHO/αⅡbβ3细胞分别为 6 0 1%和 5 5 9% ;未转染 β3的细胞感染率较低 ,CHO细胞仅为 1% ;而单转染 β3细胞介于两者之间 ( 38 7% )。结论 汉滩病毒感染率与整合素 β3表达水平密切相关 ,整合素 β3可以促进HTNV的入胞作用
Objective To investigate the relationship between integrin β3 expression and Hantaan virus (HTNV) infection. METHODS: Human integrin β3, αv and αIIb eukaryotic expression vectors were co-transfected into CHO cells respectively. The expression of exogenous genes was detected by indirect immunofluorescence assay (IFA) and flow cytometry (FCM) Stably transfected CHO cells were infected, and viral antigens were detected using IFA and FCM. Results The expression of integrin β3 was highly expressed in co-transfection group, and the target protein was mainly located on the cell membrane. Although β3 single transfection group had a certain amount of cell membrane expression, its expression level was lower than that of cotransfection group The expression of target proteins in αv and αⅡb single transfection group was significantly lower than that in co-transfection group (P <0.01), and the localization of the target protein was changed. The co-transfected cells with higher virus infection rate showed 60% and 59.5% of CHO / αvβ3 and CHO / αⅡbβ3 cells, respectively. The cell infection rate of non-transfected β3 cells was low, with only 1% of CHO cells. Transfection of β3 cells was in between (38 7%). Conclusion The infection rate of Hantaan virus is closely related to the expression of integrin β3, and integrin β3 can promote the entry of HTNV