Entry of a heparan sulphate-binding HRV8 variant strictly depends on dynamin but not on clathrin, caveolin, and flotillin
Entry of a heparan sulphate-binding HRV8 variant strictly depends on dynamin but not on clathrin, caveolin, and flotillin
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DOI：
10.1016/j.virol.2010.12.042
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发表时间：
2011-03-30
期刊：
影响因子：
3.7
通讯作者：
Blaas, Dieter
中科院分区：
文献类型：
--
作者：
Khan, Abdul Ghafoor;Pickl-Herk, Angela;Blaas, Dieter
主要组型人鼻病毒8型可通过硫酸肝素进入细胞。当HRV8内化到ICAM-1阴性的横纹肌肉瘤(RD)细胞中时，HRV8在细胞内蓄积，但在高MOI下使用3天后就会引起CPE。通过在RD和HeLa细胞之间交替进行三个盲传的适应，导致变异的HRV8v在酸性pH下稳定性降低，允许在没有ICAM-1的情况下产生感染。HRV8v在1天内以10倍低的MOI产生CPE。共聚焦荧光显微镜共定位以及药物和显性负性抑制剂的使用表明，病毒的摄取是不依赖于笼状蛋白、洞穴蛋白和Flotillin的。然而，它被王朝、阿米洛利和EIPA阻止。此外，HRV8v诱导FITC-葡聚糖摄取，并与该液相标志物共定位。在高表达ICAM-1的RD细胞中，HRV8v通过HS进入细胞的途径与HRV14相似，但可被DATORE完全抑制。(C)2010 Elsevier Inc.保留所有权利。
The major group human rhinovirus type 8 can enter cells via heparan sulphate. When internalized into ICAM-1 negative rhabdomyosarcoma (RD) cells, HRV8 accumulated in the cells but caused CPE only after 3 days when used at high MOI. Adaptation by three blind passages alternating between RD and HeLa cells resulted in variant HRV8v with decreased stability at acidic pH allowing for productive infection in the absence of ICAM-1. HRV8v produced CPE at 10 times lower MOI within 1 day. Confocal fluorescence microscopy colocalization and the use of pharmacological and dominant negative inhibitors revealed that viral uptake is clathrin, caveolin, and flotillin independent. However, it is blocked by dynasore, amiloride, and EIPA. Furthermore, HRV8v induced FITC-dextran uptake and colocalized with this fluid phase marker. Except for the complete inhibition by dynasore, the entry pathway of HRV8v via HS is similar to that of HRV14 in RD cells that overexpress ICAM-1. (c) 2010 Elsevier Inc. All rights reserved.