Characterization of West Nile viral replication and maturation in peripheral neurons in culture.
The North American West Nile virus (WNV), New York 1999 strain, appears to be highly neurotropic, and its neuroinvasiveness is an important aspect of human disease. The authors have developed an in vitro model to study WNV replication and protein processing in neurons. They compared WNV infection of the dorsal root ganglion (DRG) neurons (sensory neurons) and PC-12 cells (sympathetic neurons) to WNV infection of the mosquito cell line, C6/36, and Vero cells. WNV infection of both neuronal cell types and C6/36 cells was not cytopathic up to 30 days post infection, and continual viral shedding was observed during this period. However, WNV infection of Vero cells was lytic. Interestingly, WNV infection of neurons was not efficient, requiring a high multiplicity of infection of > or = 10. Indirect immunofluorescence assays using normal and confocal microscopy with flavivirus-reactive antibodies and WNV-infected neurons demonstrated viral antigen mostly associated with the plasma membrane and in the neurite processes. Treatment of WNV-infected C6/36, PC-12, or DRG cells with brefeldin A (BFA; a trans-Golgi inhibitor) or nocadazole (a beta-tubulin inhibitor) had little effect on viral maturation and secretion. Treatment of WNV-infected Vero cells with BFA resulted in a 1000-fold decrease in viral titer, but nocodazole had no effect. Our studies suggest that even though PC-12 and DRG neurons are mammalian cells, viral protein processing and maturation in these cells more closely resembles replication in C6/36 insect cells than in mammalian Vero cells.
Hunsperger
Elizabeth
E
Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, Department of Health and Human Services, Fort Collins, CO 80522, USA. enh4@cdc.gov
Roehrig
John T
JT
D000818:Animals; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D005727:Ganglia, Spinal / Q000821:virology; D066298:In Vitro Techniques; D018613:Microscopy, Confocal; D009474:Neurons / Q000821:virology*; D016716:PC12 Cells; D051381:Rats; D014709:Vero Cells; D014779:Virus Replication / Q000502:physiology*; D014902:West Nile virus / Q000502:physiology*