Infectious dengue-1 virus entry into mosquito C6/36 cells

• Published in: Virus research Vol. 160; no. 1-2; pp. 173 - 179
• Main Authors: Acosta, Eliana G., Castilla, Viviana, Damonte, Elsa B.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2011
• Subjects: ammonium chloride; Animals; Cell Line; chlorpromazine; clathrin; Clathrin-Coated Vesicles - virology; Clathrin-mediated endocytosis; coated vesicles; Culicidae; Dengue virus; Dengue Virus - growth & development; Dengue Virus - physiology; Dengue Virus - ultrastructure; Dengue virus type 1; Dengue virus-1; dynamins; electron microscopy; Endocytosis; fluorescent antibody technique; Microscopy, Electron; Mosquito cells; nystatin; pathogenicity; plasma membrane; serotypes; virion; Virus entry; Virus Internalization; viruses; yields; USA, Hawaii; Dengue virus-1; Mosquito cells; Clathrin-mediated endocytosis; Virus entry
• ISSN: 0168-1702; 1872-7492
• DOI: 10.1016/j.virusres.2011.06.008

• The entry of DENV-1 to mosquito cells occurs by pH and clathrin-mediated endocytosis. • Reference strains and clinical isolates of DENV-1 use the same entry route. • The four DENV serotypes infect mosquito cells by clathrin endocytic pathway. 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.