Rab8, a Vesicular Traffic Regulator, Is Involved in Dengue Virus Infection in HepG2 Cells
Rab8, a Vesicular Traffic Regulator, Is Involved in Dengue Virus Infection in HepG2 Cells
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DOI：
10.1159/000151531
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发表时间：
2008-08
期刊：
影响因子：
4.6
通讯作者：
Xiaofeng Xu;Zong-tao Chen;Jun-lei Zhang;Wei Chen;Jia-li Wang;Yan-Ping Tian;Na Gao;J. An
中科院分区：
文献类型：
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作者：
Xiaofeng Xu;Zong-tao Chen;Jun-lei Zhang;Wei Chen;Jia-li Wang;Yan-Ping Tian;Na Gao;J. An
目的：登革热病毒（DV）的发病机制尚未完全阐明。 Rab8 调节从高尔基体到质膜的囊泡运输，DV 在质膜处成熟，然后通过胞吐作用递送。在本研究中，在 HpeG2 细胞中研究了 Rab8 参与 DV 血清型 2 (DV2) 感染的情况。方法：免疫染色观察Rab8、DV2的分布及感染细胞数。构建 HepG2Rab8AM 和 HepG2Rab8DN 细胞，分别稳定表达 Rab8 的组成型活性突变体和显性失活突变体，并通过流式细胞术进行评估。通过标准空斑测定检测感染性病毒体的产生和DV2进入量。通过实时RT-PCR检测病毒RNA复制。结果：HpeG2 细胞中 Rab8 与 DV2 高度共定位，HepG2Rab8AM 和 HepG2Rab8DN 细胞中 DV 抗原阳性细胞数量减少。此外，从这些细胞中释放的子代病毒也大大减少。细胞内产生的感染性病毒颗粒也显着减少，病毒RNA复制也有不同程度的下调。此外，进入这些细胞的病毒减少了约 80%。结论：我们的数据表明Rab8的功能对于DV2感染很重要，Rab8可能参与DV2感染。
Objective: The pathogenesis of dengue virus (DV) has not been completely clarified. Rab8 regulates vesicular traffic from Golgi to plasma membrane where DV is matured and then delivered by exocytosis. In this study, involvement of Rab8 in DV serotype 2 (DV2) infection was investigated in HpeG2 cells. Methods: Distributions of Rab8 and DV2, and the number of infection cells were observed by immunostaining. HepG2Rab8AM and HepG2Rab8DN cells were constructed to stably express a constitutively active mutant of Rab8 and a dominant negative mutant, respectively, which were assessed by flow cytometry. Production of infectious virions and the amounts of DV2 entry were detected by standard plaque assay. Viral RNA replication was detected by real-time RT-PCR. Results: Rab8 showed high co-localization with DV2 in HpeG2 cells and the amount of DV antigen-positive cells decreased in HepG2Rab8AM and HepG2Rab8DN cells. Also, progeny virus released from those cells was drastically reduced. Infectious virions produced in cells were also significantly reduced, while the viral RNA replication was down-regulated by a different level. Furthermore, viral entry into those cells was reduced by about 80%. Conclusions: Our data suggest that the function of Rab8 is important for DV2 infection, and Rab8 may be involved in DV2 infection.