Infection of polarized epithelial cells with flavivirus West Nile: polarized entry and egress of virus occur through the apical surface

Abstract

Both polarized epithelial Vero (C1008) and non-polarized Vero (control) cells were grown on permeable cell culture inserts and infected either apically or basolaterally with West Nile (WN) or Kunjin (KUN) virus. KUN virus (closely related to WN virus) was used as a comparison. Using indirect immunofluorescence and plaque assays of productive virus titres, entry of WN and KUN viruses was confined to the apical surface of polarized epithelial cells. For the first time, these results provided evidence on the distribution of flavivirus-specific receptor(s) in polarized epithelial cells; that is to say that receptor expression was shown to be predominant at the apical surface. In addition, the release of these viruses from polarized Vero C1008 epithelial cells was also examined. Egress of WN virus strain Sarafend (S) was observed to occur predominantly at the apical surface of Vero C1008 cells. In contrast, the release of KUN virus was bi-directional from polarized Vero C1008 cells. Furthermore, disruption of the cellular microtubule network was shown to inhibit the apical release of WN (S) virus but had no effect on the release of KUN virus. Hence, the difference in the release of these closely related viruses suggested the involvement of a microtubule-dependent, polarized sorting mechanism for WN virus proteins but not for KUN virus proteins in polarized epithelial cells.