Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
Endophilin-A2-mediated endocytic pathway is critical for enterovirus 71 entry into caco-2 cells
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Endophilin-A2 介导的内吞途径对于肠道病毒 71 进入 caco-2 细胞至关重要
DOI：
10.1080/22221751.2019.1618686
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发表时间：
2019-05-28
影响因子：
13.2
通讯作者：
Qi, Zhong-Tian
中科院分区：
文献类型：
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作者：
Chen, Sheng-Lin;Liu, Yan-Gang;Qi, Zhong-Tian
摘要肠道病毒71型（EV 71）通常通过口-粪途径传播，并在穿过肠粘膜时引发感染。我们对它穿过肠粘膜的机制的有限理解阻碍了有效治疗选择的发展。在这里，使用RNA干扰筛选结合化学抑制剂或显性负性蛋白的过表达，我们发现EV 71进入Caco-2细胞，极化的人肠上皮细胞系，不涉及网格蛋白和小窝依赖性内吞途径或巨胞饮，但需要GTP结合蛋白发动蛋白2和细胞骨架重塑。使用靶向内啡肽家族成员的siRNA揭示了内啡肽-A2对于Caco-2细胞摄取EV 71颗粒是必需的。亚细胞分析显示，内化的EV 71病毒粒子主要与endophilin-A2共定位在细胞膜皱褶和核周区域。结合病毒进入动力学，这些数据表明EV 71主要通过内嗜蛋白A2介导的内吞（EME）途径进入Caco-2细胞。最后，我们发现，内化的EV 71病毒粒子被转运到内体分选复合物所需的运输（ESCRT）相关的多泡体（MVB）。这些数据为阻断EV 71感染提供了有吸引力的治疗靶点。
ABSTRACT Enterovirus 71 (EV71) is typically transmitted by the oral-faecal route and initiates infection upon crossing the intestinal mucosa. Our limited understanding of the mechanisms by which it crosses the intestinal mucosa has hampered the development of effective therapeutic options. Here, using an RNA interference screen combined with chemical inhibitors or the overexpression of dominant negative proteins, we found that EV71 entry into Caco-2 cells, a polarized human intestinal epithelial cell line, does not involve clathrin- and caveolae-dependent endocytic pathways or macropinocytosis but requires GTP-binding protein dynamin 2 and cytoskeleton remodelling. The use of siRNAs targeting endophilin family members revealed that endophlin-A2 is essential for the uptake of EV71 particles by Caco-2 cells. Subcellular analysis revealed that internalized EV71 virions largely colocalized with endophilin-A2 at cytomembrane ruffles and in the perinuclear area. Combined with viral entry kinetics, these data suggest that EV71 enters Caco-2 cells mainly via an endophilin-A2-mediated endocytic (EME) pathway. Finally, we showed that internalized EV71 virions were transported to endosomal sorting complex required for transport (ESCRT)-related multivesicular bodies (MVBs). These data provide attractive therapeutic targets to block EV71 infection.