Entry of human rhinovirus 89 via ICAM-1 into HeLa epithelial cells is inhibited by actin skeleton disruption and by bafilomycin
Entry of human rhinovirus 89 via ICAM-1 into HeLa epithelial cells is inhibited by actin skeleton disruption and by bafilomycin
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DOI：
10.1007/s00705-013-1797-1
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发表时间：
2014-01-01
影响因子：
2.7
通讯作者：
Blaas, Dieter
中科院分区：
文献类型：
--
作者：
Pfanzagl, Beatrix;Andergassen, Daniel;Blaas, Dieter
HRV89是一种主要的鼻病毒，使用细胞间粘附分子1（ICAM-1）进入细胞，而次要的HRV2使用LDL受体进行网格蛋白介导的内吞作用。发现HRV89进入HeLa上皮细胞是低效的，并且在与细胞孵育3小时后在质膜上仍然检测到感染性病毒。内吞作用，因此感染，HRV 89，但不是HRV 2，几乎完全阻断肌动蛋白聚合抑制剂细胞松弛素D，而磷脂酰肌醇3-激酶抑制剂LY294002没有影响感染病毒。细胞松弛素D也抑制主要组HRV感染的横纹肌肉瘤细胞表达ICAM-1时，可用于脱壳的时间被限制在30分钟。虽然胆固醇耗竭强烈抑制HRV 89感染的HeLa细胞，它只轻微影响HRV 89的内吞作用，表明脂筏环境是不是必不可少的病毒摄取。钠-质子交换抑制剂5-（N-乙基-N-异丙基）阿米洛利（EIPA）仅在也抑制HRV 2感染和Alexa 488-转铁蛋白进入的浓度下显著减少HRV 89的细胞进入和感染。这些数据排除了经典的巨胞饮作用作为HPV 89在HeLa细胞中的感染性进入途径。值得注意的是，质子ATP酶抑制剂巴弗洛霉素强烈影响两种病毒的细胞进入，这表明鼻病毒内吞作用的膜下pH值的作用。
HRV89, a major-group rhinovirus, uses intercellular adhesion molecule 1 (ICAM-1) for cell entry, while minor-group HRV2 uses the LDL receptor for clathrin-mediated endocytosis. Entry of HRV89 into HeLa epithelial cells was found to be inefficient, and infectious virus was still detected on the plasma membrane after 3 h of incubation with the cells. Endocytosis, and consequently infection, of HRV89 but not of HRV2, was almost completely blocked by the actin-polymerization inhibitor cytochalasin D, while the phosphatidylinositol 3-kinase inhibitor LY294002 had no effect on infection with either virus. Cytochalasin D also inhibited major-group HRV infection of rhabdomyosarcoma cells expressing ICAM-1 when the time available for uncoating was limited to 30 min. Although cholesterol depletion strongly inhibited HRV89 infection of HeLa cells, it only slightly affected HRV89 endocytosis, indicating that a lipid raft environment was not essential for virus uptake. The sodium-proton exchange inhibitor 5-(N-ethyl-N-isopropyl) amiloride (EIPA) significantly reduced cell entry and infection by HRV89 only at a concentration that also inhibited HRV2 infection and Alexa 488-transferrin entry. These data rule out classical macropinocytosis as an infectious entry pathway of HRV89 in HeLa cells. Notably, the proton ATPase inhibitor bafilomycin strongly affected cell entry of both viruses, suggesting a role for submembraneous pH in rhinovirus endocytosis.