Chiang Cheng-Feng, Flint Mike, Lin Jin-Mann S, Spiropoulou Christina F
Viral Special Pathogens Branch, Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
Chronic Viral Diseases Branch, Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
PLoS One. 2016 Oct 25;11(10):e0164768. doi: 10.1371/journal.pone.0164768. eCollection 2016.
Andes virus (ANDV) is the major cause of hantavirus pulmonary syndrome (HPS) in South America. Despite a high fatality rate (up to 40%), no vaccines or antiviral therapies are approved to treat ANDV infection. To understand the role of endocytic pathways in ANDV infection, we used 3 complementary approaches to identify cellular factors required for ANDV entry into human lung microvascular endothelial cells. We screened an siRNA library targeting 140 genes involved in membrane trafficking, and identified 55 genes required for ANDV infection. These genes control the major endocytic pathways, endosomal transport, cell signaling, and cytoskeleton rearrangement. We then used infectious ANDV and retroviral pseudovirions to further characterize the possible involvement of 9 of these genes in the early steps of ANDV entry. In addition, we used markers of cellular endocytosis along with chemical inhibitors of known endocytic pathways to show that ANDV uses multiple routes of entry to infect target cells. These entry mechanisms are mainly clathrin-, dynamin-, and cholesterol-dependent, but can also occur via a clathrin-independent manner.
安第斯病毒（ANDV）是南美洲汉坦病毒肺综合征（HPS）的主要病因。尽管病死率很高（高达40%），但目前尚无获批用于治疗ANDV感染的疫苗或抗病毒疗法。为了解内吞途径在ANDV感染中的作用，我们采用了3种互补方法来鉴定ANDV进入人肺微血管内皮细胞所需的细胞因子。我们筛选了一个针对140个参与膜运输基因的小干扰RNA文库，鉴定出55个ANDV感染所需的基因。这些基因控制主要的内吞途径、内体运输、细胞信号传导和细胞骨架重排。然后，我们使用感染性ANDV和逆转录病毒假病毒，进一步表征其中9个基因在ANDV进入早期步骤中可能的参与情况。此外，我们使用细胞内吞标记物以及已知内吞途径的化学抑制剂，来表明ANDV利用多种进入途径感染靶细胞。这些进入机制主要依赖网格蛋白、发动蛋白和胆固醇，但也可以通过非网格蛋白依赖的方式发生。
