Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
Virology. 2010 May 25;401(1):18-28. doi: 10.1016/j.virol.2010.02.015. Epub 2010 Mar 3.
Ebola virus (EBOV) infects several cell types and while viral entry is known to be pH-dependent, the exact entry pathway(s) remains unknown. To gain insights into EBOV entry, the role of several inhibitors of clathrin-mediated endocytosis in blocking infection mediated by HIV pseudotyped with the EBOV envelope glycoprotein (EbGP) was examined. Wild type HIV and envelope-minus HIV pseudotyped with Vesicular Stomatitis Virus glycoprotein (VSVg) were used as controls to assess cell viability after inhibiting clathrin pathway. Inhibition of clathrin pathway using dominant-negative Eps15, siRNA-mediated knockdown of clathrin heavy chain, chlorpromazine and sucrose blocked EbGP pseudotyped HIV infection. Also, both chlorpromazine and Bafilomycin A1 inhibited entry of infectious EBOV. Sensitivity of EbGP pseudotyped HIV as well as infectious EBOV to inhibitors of clathrin suggests that EBOV uses clathrin-mediated endocytosis as an entry pathway. Furthermore, since chlorpromazine inhibits EBOV infection, novel therapeutic modalities could be designed based on this lead compound.
埃博拉病毒(EBOV)感染多种细胞类型,尽管已知病毒进入是依赖 pH 值的,但确切的进入途径仍不清楚。为了深入了解 EBOV 的进入机制,研究了几种网格蛋白介导的内吞作用抑制剂在阻断 HIV 假型感染中的作用,该 HIV 假型由 EBOV 包膜糖蛋白(EbGP)假型化。使用野生型 HIV 和包膜缺失的假型化 HIV 来评估氯丙嗪和蔗糖抑制感染性 EBOV 进入的能力,这些 HIV 假型化用 Vesicular Stomatitis Virus glycoprotein (VSVg) 进行假型化,作为评估细胞活力的对照。使用显性负性 Eps15 抑制网格蛋白途径、氯丙嗪和蔗糖抑制 clathrin 重链的 siRNA 敲低、氯丙嗪和 Bafilomycin A1 均能阻断 EbGP 假型 HIV 感染。此外,EbGP 假型 HIV 以及感染性 EBOV 对网格蛋白抑制剂敏感,表明 EBOV 利用网格蛋白介导的内吞作用作为进入途径。此外,由于氯丙嗪能抑制 EBOV 感染,因此可以基于该先导化合物设计新的治疗方法。