title: Infectious dengue-1 virus entry into mosquito C6/36 cells.
abstract: 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-β-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.
authors.affiliation: Laboratorio de Virología, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pabellón 2, Piso 4, 1428 Buenos Aires, Argentina.
authors.forename: Eliana G
authors.initials: EG
authors.lastname: Acosta
authors.forename: Viviana
authors.initials: V
authors.lastname: Castilla
authors.forename: Elsa B
authors.initials: EB
authors.lastname: Damonte
mesh_terms: D000818:Animals; D002460:Cell Line; D022163:Clathrin-Coated Vesicles / Q000821:virology; D009033:Culicidae; D003716:Dengue Virus / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D004705:Endocytosis; D008854:Microscopy, Electron; D053586:Virus Internalization