水泡性口炎病毒与Monocyte及MoDC的互作研究

Study on Interactions of Vesicular Stomatitis Virus with Monocyte and MoDC

为了探究水泡性口炎病毒（ Vesicular stomatitis virus, VSV）与天然宿主免疫细胞的互作关系,实验用M蛋白第51位甲硫氨酸残基缺失后的重组病毒（ VSVAM 51-GFP ）和第51位甲硫氨酸敲除、第 221位缬氨酸突变为 苯丙氨酸、第226位丝基酸突变为精氨酸的重组病毒（ VSV-MT-GFP ）以及野生型重组病毒（VSV-GFP）感染猪单核 细胞（ Monocyte）和单核细胞诱导的树突状细胞（ Monocyte-dcriveddendritic cell, MoDC）,通过空斑实验制作病毒生 长曲线,用激光共聚焦显微镜和流式细胞仪检测细胞的感染状况, E LISA 检测细胞培养上清中的细胞因子表达变 化.结果表明, VSV、 VSVAM 51、 VSV-MT 均可感染 Monocyte和 MoDC,且 Monocyte的感染率仅为8.38%.3 种病 毒在 MoDC中可以大量复制,但无法在 Monocyte中扩增.且与野生型VSV相比, VSViM51、VSV-MT 细胞致病性 依次减弱.VSV能有效激活TNF -α、 IL-8、 FN-α 相关免疫细胞因子在MoDC中的表达,且 M 蛋白突变程度越大,病毒激活能力越强.

In order to explore the interactions between VSV(Vesicular stomatitis virus)and immune cells of natural host,three recombinant VSVs,including the wild-type VSV-GFP,the VSV with M51deletion(VSV_ΔM51-GFP)and the VSV with triple mutations of M51 deletion,V221Fand S226R(VSVM_T-GFP),were used to infect the pig monocyte and monocyte-derived dendritic cell.The plaque experiment was done to make virus growth curve.Then confocal laser scanning microscope and Flow cytometry were used to assess the cell infection status.The change in cytokines expression in MoDC was detected by ELISA assay kit.The results show that three viruses can infect monocyte and MoDC,and virus infection rate of Monocyte is only 8.38%.The three viruses are unable to replicate in quantity in monocyte,but cannot proliferate in monocyte.Compared with wild-type VSV,the pathogenicity of VSV_ΔM51and VSV-M_T decreases successively.They can effectively activate the expression of TNF-α,IL-8,IFN-αin MoDC,and the virus activation capacity is positively correlated with the degree of M protein mutation.