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目的检测CHO细胞中稳定表达人整合素β3对登革2型病毒(Dengue virus serotype 2,DV2)感染的影响。方法将表达人整合素β3的质粒转染CHO细胞,通过G418筛选、流式细胞学检测和间接免疫荧光实验,建立稳定表达整合素β3的CHO细胞株;利用该细胞株进行感染实验,检测人整合素β3稳定表达对DV2易感性的影响。结果共获得了C9、C10、F8和H6 4个高表达整合素β3的细胞克隆,其中克隆F8中整合素β3的表达水平最高,是对照组的1.89倍。感染实验结果显示,进入整合素β3高表达细胞株的病毒数量均有所增高,增高的比例与整合素β3的表达增高趋势基本一致,进入F8细胞株的病毒量最高,是对照细胞的2.63倍,说明整合素β3有助于DV2进入细胞。结论在CHO细胞中外源性表达整合素β3可以提高DV2的感染率。
Objective To detect the effect of stably expressing human integrin β3 on the infection of Dengue virus serotype 2 (DV2) in CHO cells. Methods CHO cells expressing human integrin β3 were transfected into CHO cells. CHO cells stably expressing integrin β3 were established by G418 selection, flow cytometry and indirect immunofluorescence assay. The cell lines were used to infect CHO cells. Effect of Integrin β3 Stable Expression on DV2 Susceptibility. Results Four clones of C9, C10, F8 and H6 highly expressed integrin β3 were obtained. The expression level of integrin β3 in clone F8 was the highest, which was 1.89 times that of the control group. The results of infection showed that the number of viruses entering into integrin β3 high-expressing cell lines increased, the increasing proportion was consistent with the increasing trend of integrin β3, the highest viral load entering F8 cell line was 2.63 times that of control cells , Indicating that integrin beta 3 helps DV2 enter cells. Conclusion The exogenous expression of integrin β3 in CHO cells can increase the infection rate of DV2.