【摘要】 目的探讨树突状细胞特异性细胞间黏附分子3结合非整合素(dendritic cell specific intercellular adhesion molecule-3 grabbing non integrin,DC-SIGN)的表达对肠道病毒71型(enterovirus type 71,EV71)体外感染人树突状细胞(dendritic cells, DCs)和293T细胞的影响。方法梯度密度离心法分离人外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),体外诱导培养DCs,通过RT-PCR方法克隆DC-SIGN分子,并构建稳定表达DC-SIGN分子的载体pLB-DC-SIGN,脂质体方法转染293T细胞。分别用EV71感染对照组293T细胞和过表达DC-SIGN的293T细胞,荧光定量PCR检测病毒mRNA,蛋白质印迹法检测EV71/VP1的表达水平;同时用DC-SIGN抑制剂酵母甘露聚糖阻断DCs表面的DC-SIGN分子后,荧光定量PCR检测病毒mRNA,蛋白质印迹法检测EV71/VP1的表达水平。结果过表达DC-SIGN的293T细胞与对照组293T细胞相比,感染病毒量明显增高,EV71/VP1表达量显著增加,而阻断DC-SIGN的DCs病毒感染量低于对照组DCs,EV71/VP1表达量明显降低,差异均有统计学意义(P<0.05)。结论 DC-SIGN分子可促进EV71体外感染DCs和293T细胞,可能是EV71感染的受体之一。更多还原

【Abstract】 Objective To investigate the influence of dendritic cell specific intercellular adhesion molecule-3 grabbing non integrin（DC-SIGN） expression on the infection of enterovirus type 71（EV71） for dendritic cells（DCs） and 293 T cells. Methods Peripheral blood mononuclear cells（PBMCs） were isolated from human peripheral blood by gradient density centrifugation and induced to DCs. The DC-SIGN gene was amplified by polymerase chain reaction（PCR） for construction of transient expression vector pLB-DC-SIGN which was then transfected into 293 T cells using Lipofectamine^TM 2000. After 293 T cell and 293 T-pLB-DC-SIGN was infected with EV71, EV71 mRNA was identified by fluorescence quantitative PCR. EV71/VP1 was identified by western blot. After the expression of DC-SIGN on DCs was blocked by yeast mannan, EV71 mRNA was identified by fluorescence quantitative PCR and EV71/VP1 was identified by western blot. Results EV71 viral load and EV71/VP1 expression level in 293 T cells overexpressed DC-SIGN was significantly higher than that of control 293 T, while the viral load and EV71/VP1 expression level in DCs blocked with DC-SIGN inhibitor was lower than that of control DCs（P<0.05）. Conclusion DC-SIGN may be one of the receptors of EV71 who promote EV71 infection in DCs and 293 T cells in vitro.更多还原