Alternative infectious entry pathways for dengue virus serotypes into mammalian cells
Alternative infectious entry pathways for dengue virus serotypes into mammalian cells
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DOI：
10.1111/j.1462-5822.2009.01345.x
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发表时间：
2009-10-01
影响因子：
3.4
通讯作者：
Damonte, Elsa B.
中科院分区：
文献类型：
--
作者：
Acosta, Eliana G.;Castilla, Viviana;Damonte, Elsa B.
采用生化抑制剂、胞吞途径蛋白质显性失活突变体、荧光显微镜和感染性测定等方法，对两种登革病毒（DENV）血清型进入Vero细胞的情况进行了分析。通过用丹磺尸胺和氯丙嗪处理以及Eps15蛋白的显性负性形式的过表达，证明了用于生产性DENV-1内化到Vero细胞中的网格蛋白介导的内吞作用，而DENV-2在相同细胞系统中的感染性进入是独立于网格蛋白的。用抑制剂制霉菌素和甲基-β-环糊精处理，以及用显性阴性小窝蛋白-1转染Vero细胞，对DENV-2病毒感染没有影响。通过使用K44A突变体和抑制剂dynasore，还显示发动蛋白是DENV-2进入所需的。因此，DENV-2感染进入Vero细胞是通过非经典的内吞途径发生的，不依赖于网格蛋白、小窝和脂筏，但依赖于发动蛋白。相反，DENV-2进入A549细胞是网格蛋白依赖性的，如先前在HeLa、C6/36和BS-C-1细胞中报道的。我们的研究结果第一次最终显示了DENV-1和DENV-2感染进入共同宿主细胞Vero细胞的差异模式，以及给定血清型DENV-2进入不同类型细胞的替代进入途径。
P>The entry of two dengue virus (DENV) serotypes into Vero cells was analysed using biochemical inhibitors, dominant negative mutants of cellular proteins involved in endocytic pathways, fluorescence microscopy and infectivity determinations. By treatment with dansylcadaverine and chlorpromazine and overexpression of a dominant negative form of the Eps15 protein, a clathrin-mediated endocytosis for productive DENV-1 internalization into Vero cells was demonstrated whereas the infectious entry of DENV-2 in the same cell system was independent of clathrin. Treatment with the inhibitors nystatin and methyl-beta-cyclodextrin, as well as transfection of Vero cells with dominant negative caveolin-1, had no effect on DENV-2 virus infection. It was also shown, by using the K44A mutant and the inhibitor dynasore, that dynamin was required for DENV-2 entry. Consequently, the infectious entry of DENV-2 into Vero cells occurs by a non-classical endocytic pathway independent of clathrin, caveolae and lipid rafts, but dependent on dynamin. By contrast, DENV-2 entry into A549 cells was clathrin-dependent, as previously reported in HeLa, C6/36 and BS-C-1 cells. Our results conclusively show, for the first time, a differential mode of infective entry for DENV-1 and DENV-2 into a common host cell, Vero cells, as well as alternative entry pathways for a given serotype, DENV-2, into different types of cells.