Entry mechanisms of visualized pseudotype virus bearing the envelope proteins of hepatitis C virus.

丙型肝炎病毒的受体已经确定，但丙型肝炎病毒进入靶细胞的途径还不完全确定。作为一种替代病毒系统，以水疱性口炎病毒(VSV)和逆转录病毒或细胞培养的丙型肝炎病毒(HCVcc)为基础，发展了瞬时携带丙型肝炎病毒包膜蛋白的伪型病毒。在这里，我们试图开发一种荧光标记的伪型VSV或重组VSV来可视化研究进入机制。此外，为了研究丙型肝炎病毒感染激活下游信号通路的机制，我们利用含有丙型肝炎病毒囊膜蛋白(HCVpv)的假型水疱性口炎病毒和JFH-1病毒与药物抑制剂相结合。我们报道了丙型肝炎病毒利用磷脂酶C(PLC)/蛋白激酶C(PKC)依赖的信号进入。肌动蛋白解聚试剂抑制肌动蛋白重排可显著减少HCVpv感染。总之，这些结果表明，激活PLC和PKC依赖的通路是进入丙型肝炎病毒所必需的，并参与设定病毒内吞的肌动蛋白重排。

The receptors of HCV have been characterized, yet the entry pathways of the HCV into target cells are incompletely defined. As a surrogate virus system, pseudotype viruses transiently bearing HCV envelope proteins based on the vesicular stomatitis virus (VSV) and retrovirus or cell cultured HCV (HCVcc) have been developed. Here, we have tried to develop a fluorescence-labeled pseudotype VSV or recombinant VSV for a visualized investigation of the entry mechanisms. Furthermore, to characterize the mechanisms by which HCV infection activates downstream signaling pathways, we utilized a pseudotype vesicular stomatitis virus possessing HCV envelope proteins (HCVpv) and JFH-1 virus in combination with pharmacological inhibitors. We reported that HCV utilizes the phospholipase C (PLC) / protein kinase C (PKC)-dependent signaling for the entry. Inhibition of actin rearrangement by actin-depolymerizing reagents markedly diminishes HCVpv infection. Overall, these results indicate that the activation of PLC and PKC-dependent pathway is required for the entry of HCV and is involved in setting the actin rearrangement for the endocytosis of the virus.