A 65-kDa trypsin-sensible membrane cell protein as a possible receptor for dengue virus in cultured neuroblastoma...

Dengue virus infects primary neurons in mouse experimental model and tissue culture cells of the central nervous system (CNS). In the present work, a mouse neuroblastoma cell line (N1E-115) and a human neuroblastoma cell line (SK-N-SH), susceptible to dengue virus infection were used to study the presence of cell membrane receptor for dengue-2. By day 5 postinfection (pi), viral antigen was detected by immunofluorescence in the cytoplasm and surrounding the nucleus of N1E-115 cells, while on day 7 pi, it was also present along neural extensions. Infection of N1E-115 cells was diminished with trypsin treatment but not with neuraminidase or endoglycosidase H. Partially purified cell membrane proteins from neuroblastoma cells were analyzed by the Virus Overlay Protein Blot Assay (VOPBA), and a single band migrating at 65 kDa was detected in mouse and human neuroblastoma cells but not in C6, a non-susceptible rat glial cell line which was included as a negative control. The 65 kDa protein was eliminated only when nitrocellulose membranes were treated with trypsin. Analysis of neuronal cell infection by dengue virus provides a useful tool to understand the nature of cellular receptors and mechanisms involved in the infection of the nervous system by dengue viruses.