Infectious dengue-1 virus entry into mosquito C6/36 cells
Infectious dengue-1 virus entry into mosquito C6/36 cells
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DOI：
10.1016/j.virusres.2011.06.008
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发表时间：
2011-09-01
期刊：
影响因子：
5
通讯作者：
Damonte, Elsa B.
中科院分区：
文献类型：
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作者：
Acosta, Eliana G.;Castilla, Viviana;Damonte, Elsa B.
采用多种生化抑制剂结合电镜技术，分析了夏威夷登革热病毒1型（DENV-1）进入蚊虫C6/36细胞的情况。通过感染性滴定测定，氯化铵、氯丙嗪、丹西卡弗碱和达摩可抑制病毒产量，而制霉菌素和甲基- β -环糊精则没有任何影响。通过免疫荧光染色检测内化病毒粒子，证实了网格蛋白和动力蛋白抑制剂对DENV-1进入的影响。此外，电子显微照片显示，进入的病毒粒子分别附着在电子密集的质膜内陷上和包裹的囊泡内，这些囊泡分别类似于网格蛋白包裹的凹坑和囊泡。最近爆发的临床分离株对网格蛋白和动力蛋白抑制剂的敏感性与适应细胞培养的参考菌株相当。同样，DENV-3株H87和DENV-4株8124在氯化铵、氯丙嗪和丁酮的存在下也被抑制，从而得出结论，DENV血清型通过低ph依赖性网格蛋白介导的内吞作用进入蚊子细胞。(C) 2011 Elsevier BM。版权所有。
The entry of dengue virus-1 (DENV-1) strain Hawaii into mosquito C6/36 cells was analyzed using a variety of biochemical inhibitors together with electron microscopy. The treatment with ammonium chloride, chlorpromazine, dansylcadaverine and dynasore inhibited virus yields, determined by infectivity titrations, whereas nystatin and methyl-beta-cyclodextrin did not have any effect. The effect of the clathrin and dynamin inhibitors on DENV-1 entry was corroborated by detection of internalized virions using immunofluorescence staining. Furthermore, electron micrographs showed the incoming virions attached to electron-dense invaginations of the plasma membrane and within coated vesicles that resembled clathrin-coated pits and vesicles, respectively. The susceptibility to clathrin and dynamin inhibitors of clinical isolates from recent outbreaks was comparable to that shown by the cell culture-adapted reference strain. Similarly, DENV-3 strain H87 and DENV-4 strain 8124 were also inhibited in the presence of ammonium chloride, chlorpromazine and dynasore, allowing conclude that the infectious entry of DENV serotypes to mosquito cells occurs by low pH-dependent clathrin-mediated endocytosis. (C) 2011 Elsevier BM. All rights reserved.