S-phase-dependent enhancement of dengue virus 2 replication in mosquito cells, but not in human cells
S-phase-dependent enhancement of dengue virus 2 replication in mosquito cells, but not in human cells
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DOI：
10.1128/jvi.79.21.13218-13230.2005
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发表时间：
2005-11-01
影响因子：
5.4
通讯作者：
Harris, E
中科院分区：
文献类型：
--
作者：
Helt, AM;Harris, E
登革病毒（DEN）是人类节肢动物传播的病毒性疾病的最普遍原因。我们确定了细胞生长状态对蚊子和人类细胞中DEN2型病毒复制的影响，基于对细胞生长状态的操纵将有助于识别病毒和细胞决定因素的生产感染。低传代DEN2分离物感染密度阻滞细胞和循环循环的C6/36白纹伊蚊细胞的比较显示，循环循环的细胞产生更高的病毒滴度。当C6/36细胞通过thymidine （THY）阻滞在s期时，低传代DEN2分离株和高传代菌株16681的滴度分别提高了约30倍和10倍。此外，在thy处理的细胞中，病毒释放比在异步循环细胞中更早。吸附、进入、基因组剥膜和翻译不是s期C6/36细胞中病毒滴度增加的原因。与病毒滴度增加30倍相反，细胞内病毒RNA水平增加了约2倍，这表明s期反应步骤在DEN2复制周期的后期。从细胞释放的病毒RNA和蛋白质分析表明，增强的DEN2组装在很大程度上是s期产生的病毒滴度增加的原因。与C6/36细胞相比，s期人肝癌细胞或原代人成纤维细胞的DEN2滴度没有升高。这些结果证明了DEN2对蚊子和人类细胞周期的不同反应，并为详细研究介导病毒组装的机制提供了框架。
Dengue virus (DEN) is the most prevalent cause of arthropod-borne viral illness in humans. We determined the influence of cellular growth state on DEN type 2 (DEN2) replication in mosquito and human cells, based on the hypothesis that manipulation of cellular growth state will facilitate identification of viral and cellular determinants of productive infection. Comparison of density-arrested and cycling C6/36 Aedes albopictus cells infected with a low-passage DEN2 isolate revealed that cycling cells generated higher virus titers per cell. When C6/36 cells were stalled in S-phase via a thymidine (THY) block, titers of low-passage DEN2 isolates and a high-passage strain, 16681, were increased approximately 30-fold and 10-fold, respectively. Moreover, virus release was earlier in THY-treated cells than in asynchronously cycling cells. Adsorption, entry, genome uncoating, and translation were not responsible for increased titers of virus from S-phase C6/36 cells. In contrast to the 30-fold increase in virus titers, intracellular levels of viral RNA were increased approximately 2-fold, suggesting that the S-phase-responsive step is late in the DEN2 replication cycle. Analysis of viral RNA and protein released from the cells indicated that enhanced DEN2 assembly is largely responsible for increased virus titers produced during S-phase. In contrast to C6/36 cells, DEN2 titers from S-phase human hepatoma cells or primary human fibroblasts were not increased. These results demonstrate a differential response of DEN2 to the mosquito and human cell cycle and provide a framework for detailed studies into the mechanisms mediating virus assembly.