微管骨架在登革病毒感染ECV304细胞中作用的初步研究

目的研究登革病毒在与ECV304细胞相互作用过程中细胞微管骨架的变化,以及微管在病毒释放过程中的作用.方法用2型登革病毒感染ECV304细胞株,间接免疫荧光双染色技术观察微管和病毒抗原的分布;应用colcemid破坏微管骨架,检测细胞内外病毒滴度的变化,以明确微管是否参与病毒的复制.结果登革病毒感染后ECV304细胞微管骨架的排列发生明显的变化,表现为3种类型:无序排列;围绕核形成环状结构;微管结成束状形成线状突触.间接免疫荧光双染色结果显示登革病毒抗原与微管共染,分布在微管组织中心.感染后8 h,药物组上清中的病毒滴度高于一般感染组,而细胞内的病毒滴度低于一般感染组.结论登革病毒感染引起ECV304细胞微管的排列发生变化,可能是病毒感染造成了微管解聚,后者促进病毒的释放;药物的添加,加剧了对微管的破坏程度,从而进一步促进了病毒的释放.

Role of microtubules in dengue virus infection of ECV304 cell line

Objective To analyze structural features of microtubules (MT) in ECV304 cell line after infection of dengue virus serotype 2 (DV-2) and investigate the role of MT in DV life cycles in the cells. Methods After ECV304 cells were infected with DV-2, viral antigen and MT were detected by double immunostaining. Colcemid, which disrupts MT, was added while DV began to release. The viral titer was measured by plaque assay. Results That MT randomly oriented in the cytoplasm and lost normal structure, MT ring around nucleus and MT bundle was observed during infection. Localization of DV-2 antigens was seen in the vicinity of MT aster. In the group treated with colcemid, the levels of extracellular and intracellular virus were higher than those in the group with infection alone. Conclusion DV-2 infection induced disruption of MT cytoskeleton and the disruption could enhance viral spreading.