Endocytic pathway followed by dengue virus to infect the mosquito cell line C6/36 HT
Endocytic pathway followed by dengue virus to infect the mosquito cell line C6/36 HT
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DOI：
10.1016/j.virol.2008.05.012
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发表时间：
2008-08-15
期刊：
影响因子：
3.7
通讯作者：
del Angel, Rosa M.
中科院分区：
文献类型：
--
作者：
Mosso, Clemente;Galvan-Mendoza, Ivan J.;del Angel, Rosa M.
分析了登革热病毒感染蚊虫细胞C6/36HT所遵循的内吞途径。使用Dil标记的病毒粒子和实时成像，确定病毒进入C6/36 HT大约需要5到7分钟。用蔗糖和巴菲霉素A预处理C6/36HT细胞，但不用非利培平，可抑制登革病毒感染高达80%。此外，Eps15显性-负性突变体的过度表达使登革热感染减少了50%，这表明登革热病毒是通过笼蛋白介导的内吞作用进入的，并且是pH依赖的。免疫荧光双标记法显示，DIL标记颗粒在感染后5min与早期内体共定位，在30min主要与溶酶体共定位。最后，诺可达唑和细胞松弛素D破坏微管和微丝可使病毒感染减少80%以上。综上所述，这些结果表明，登革热病毒粒子通过网状蛋白介导的内吞作用进入C6/36 HT细胞，在病毒RNA释放到细胞质之前，通过内体途径从早期的内吞体到酸性溶酶体。(C)2008 Elsevier Inc.保留所有权利。
The endocytic pathway followed by dengue virus to infect the mosquito cells C6/36 HT was analyzed. Using DIL-labeled virions and real-time imaging it was determined that viral entry into C6/36 HT takes approximately 5 to 7 min. Pretreatment of C6/36 HT cells with sucrose and bafilomycin A, but not filipin, inhibited dengue virus infection up to 80%. Furthermore, the overexpression of dominant-negative mutants of Eps15, a molecule required for the formation of clathrin-coated vesicles, reduced dengue infection up to 50%, indicating that dengue Virus entry is through clathrin-mediated endocytosis and is pH-dependent. By double-immunofluorescence assays, DIL-labeled particles were colocalized with early endosomes at 5 min and with lysosomes mainly at 30 min post-infection. Finally, disruption of the microtubule and microfilaments by nocodazole and by cytochalasin D reduced viral infection by more than 80%. Taken together these results indicate that dengue virions enter into C6/36 HT cells by clathrin-mediated endocytosis, using the endosomal pathway from early endosomes to acidic lysosomes before viral RNA is released into the cytoplasm. (c) 2008 Elsevier Inc. All rights reserved.