title: Entry of dengue virus serotype 2 into ECV304 cells depends on clathrin-dependent endocytosis, but not on caveolae-dependent endocytosis.
abstract: Caveolae- and clathrin-mediated endocytosis are major internalization pathways used by several pathogens; however, their distinctive roles in dengue virus (DV) entry have not been addressed. In this study, we compared the involvement of caveolae- and clathrin-mediated endocytosis in the infectious entry of DV serotype 2 (DV2) into human endothelial-like ECV304 cells. Confocal microscopy study on DV2-infected cells showed that viral antigens were co-localized with clathrin heavy chains, epidermal growth factor pathway substrate clone 15 (Eps15), and adaptin-alpha, but not with caveolin-1. Treatment with chlorpromazine, which inhibits clathrin-dependent endocytosis, led to reduced virus entry into cells, whereas treatment with nystatin, a caveolae inhibitory agent, did not. Furthermore, gene silencing of Eps15 resulted in an average of 75% reduced infection of ECV304 cells by DV2. Our results demonstrated that DV2 enters ECV304 cells by clathrin-dependent endocytosis, not by caveolae-dependent endocytosis.
authors.affiliation: Department of Microbiology, Third Military Medical University, Chongqing, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.forename: Jia-Li
authors.initials: JL
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Jun-Lei
authors.initials: JL
authors.lastname: Zhang
authors.forename: Na
authors.initials: N
authors.lastname: Gao
authors.forename: Zong-Tao
authors.initials: ZT
authors.lastname: Chen
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Xu
authors.forename: Dong-Ying
authors.initials: DY
authors.lastname: Fan
authors.forename: Jing
authors.initials: J
authors.lastname: An
mesh_terms: D021941:Caveolae / Q000502:physiology*; D002460:Cell Line; D002966:Clathrin / Q000378:metabolism*; D003715:Dengue / Q000378:metabolism / Q000503:physiopathology* / Q000821:virology; D003716:Dengue Virus / Q000502:physiology*; D004705:Endocytosis; D006801:Humans; D053586:Virus Internalization
chemical_list: D002966:Clathrin