title: Flavivirus entry into cultured mosquito cells and human peripheral blood monocytes.
abstract: The entry modes of Japanese encephalitis (JE) and dengue-2 (DEN-2) viruses into C6/36 mosquito cells and of DEN-2 virus into human peripheral blood monocytes in vitro were studied. Inoculation of either JE or DEN-2 virions into C6/36 cells resulted in direct penetration of the virions into the cytoplasm at the cell surface in 3 stages. At stage 1, virions attached to the plasma membrane of host cells by their envelope spikes; at stage 2, the virion envelopes approximated to and eventually overlapped the host plasma membrane, and in the process the plasma membrane at the attachment sites dissolved; and, at stage 3, virions penetrated into the cytoplasm through the plasma-membrane disruptions created at the adsorption sites. Virions themselves apparently disintegrated at or near the penetration sites, for no virions were seen in the deeper cytoplasm. Coated pits did not form at the virion attachment sites, and virion-containing vesicles were not found in the cytoplasm. In the entry of DEN-2 virus into human peripheral blood monocytes, virions were found, adsorbed onto the external surface of the plasma membrane and attached to the luminal surface of macropinocytic vacuolar membranes. The latter apparently occurred as the result of ruffling and macropinocytic activities of the cells. At both sites virions penetrated into the cytoplasm through the plasma or vacuolar membrane in the same manner as they did through the plasma membrane of C6/36 cells. No evidence of viral entry by receptor-mediated endocytosis was observed. Implications of the entry mode of the mosquito cell-generated DEN-2 virus into human peripheral blood monocytes to an early process of natural, mosquito-transmitted infection is discussed.
authors.affiliation: Department of Ultrastructural Studies, Walter Reed Army Institute of Research, Washington, D.C.
authors.forename: T
authors.initials: T
authors.lastname: Hase
authors.forename: P L
authors.initials: PL
authors.lastname: Summers
authors.forename: K H
authors.initials: KH
authors.lastname: Eckels
mesh_terms: D000327:Adsorption; D000818:Animals; D002213:Capsid / Q000502:physiology; D002462:Cell Membrane / Q000382:microbiology; D002478:Cells, Cultured; D009033:Culicidae; D003716:Dengue Virus / Q000502:physiology* / Q000648:ultrastructure; D004664:Encephalitis Virus, Japanese / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D007158:Immunologic Techniques; D007700:Kinetics; D009000:Monocytes / Q000382:microbiology