Entry into and production of the Japanese encephalitis virus from C6/36 cells.
OBJECTIVE
The mosquito-borne Japanese encephalitis virus is a leading cause of encephalitis worldwide. However, few studies have investigated the kinetics of Japanese encephalitis virus internalization and production in mosquito cells, and fewer still have investigated the nature of the molecules involved in the binding of the virus to mosquito cells.


METHODS
Using the Aedes albopictus/Stegomyia albopicta-derived C6/36 cell line, Japanese encephalitis virus internalization and production were assayed by standard plaque assay, and virus binding was investigated through preinfection enzymatic treatment of cells and virus overlay protein-binding assay of membrane fractions in native and denaturing gels.


RESULTS
The internalization of the virus was nonlinear, and intracellular infective viruses were detected 8 h after infection and exported to the medium 10 h after infection. The internalization of the virus was primarily mediated by protein elements, and several bands were observed after overlay assay to membrane proteins, although mass spectroscopy was unable to identify candidate proteins. Soluble laminin produced a marginal, but dose-dependent inhibition of infection.


CONCLUSIONS
These results suggest that the mechanism of Japanese encephalitis entry, production, attachment and receptor usage are distinct from those employed by the dengue viruses.
Boonsanay
Verawan
V
Molecular Pathology Laboratory, Institute of Molecular Biology and Genetics, Mahidol University, Nakorn Pathom, Thailand.
Smith
Duncan R
DR
D000330:Aedes; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000821:virology; D003593:Cytoplasm / Q000821:virology; D004664:Encephalitis Virus, Japanese / Q000502:physiology*; D007797:Laminin / Q000502:physiology; D013058:Mass Spectrometry; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification; D011991:Receptors, Virus / Q000032:analysis / Q000302:isolation & purification; D013997:Time Factors; D010948:Viral Plaque Assay; D053585:Virus Attachment; D053586:Virus Internalization
D007797:Laminin; D011506:Proteins; D011991:Receptors, Virus