Ebola Virus Enters Host Cells by Macropinocytosis and Clathrin-Mediated Endocytosis
Ebola Virus Enters Host Cells by Macropinocytosis and Clathrin-Mediated Endocytosis
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DOI：
10.1093/infdis/jir326
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发表时间：
2011-11-01
影响因子：
6.4
通讯作者：
Schnittler, Hans-Joachim
中科院分区：
文献类型：
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作者：
Aleksandrowicz, Paulina;Marzi, Andrea;Schnittler, Hans-Joachim
病毒进入宿主细胞是感染的第一步，也是致病性的关键决定因素。在这里，我们展示了由糖蛋白GP(1，2)和基质蛋白VP40组成的埃博拉病毒样颗粒(EBOV-VLP)利用巨噬细胞吞噬和笼蛋白介导的内吞作用进入细胞。将EBOV-VLP应用于宿主细胞可诱导肌动蛋白驱动的褶皱，并增强FITC-葡聚糖摄取，这表明巨噬细胞吞噬是主要的进入机制。通过肌动蛋白聚合抑制剂(Latrunculin A)、Na(+)/H(+)-交换器(EIPA)和PI3-激酶(Wortmannin)抑制进入进一步支持了这一点。然而，部分EBOV-VLP与胞蛋白重链(Chathrin Heavy Chain，CHC)共定位，CHC小干扰RNA转染和显性负动力蛋白II-K44A突变体的表达降低了VLP的摄取。相反，我们没有发现EBOV-VLP通过小凹进入细胞的证据。这项工作确定了巨噬细胞吞噬作用是EBOV颗粒的主要途径，而依赖于笼蛋白的内吞作用是EBOV颗粒的替代进入途径。因此，EBOV似乎利用不同的进入途径，这取决于细胞类型和病毒颗粒大小。
Virus entry into host cells is the first step of infection and a crucial determinant of pathogenicity. Here we show that Ebola virus-like particles (EBOV-VLPs) composed of the glycoprotein GP(1,2) and the matrix protein VP40 use macropinocytosis and clathrin-mediated endocytosis to enter cells. EBOV-VLPs applied to host cells induced actin-driven ruffling and enhanced FITC-dextran uptake, which indicated macropinocytosis as the main entry mechanism. This was further supported by inhibition of entry through inhibitors of actin polymerization (latrunculin A), Na(+)/H(+)-exchanger (EIPA), and PI3-kinase (wortmannin). A fraction of EBOV-VLPs, however, colocalized with clathrin heavy chain (CHC), and VLP uptake was reduced by CHC small interfering RNA transfection and expression of the dominant negative dynamin II-K44A mutant. In contrast, we found no evidence that EBOV-VLPs enter cells via caveolae. This work identifies macropinocytosis as the major, and clathrin-dependent endocytosis as an alternative, entry route for EBOV particles. Therefore, EBOV seems to utilize different entry pathways depending on both cell type and virus particle size.