A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells

Abstract

Chikungunya virus (CHIKV) has resulted in several outbreaks in the past six decades. The clinical symptoms of Chikungunya infection include fever, skin rash, arthralgia, and an increasing incidence of encephalitis. The re-emergence of CHIKV with more severe pathogenesis highlights its potential threat on our human health. In this study, polarized HBMEC, polarized Vero C1008 and non-polarized Vero cells grown on cell culture inserts were infected with CHIKV apically or basolaterally. Plaque assays, viral binding assays and immunofluorescence assays demonstrated apical entry and release of CHIKV in polarized HBMEC and Vero C1008. Drug treatment studies were performed to elucidate both host cell and viral factors involved in the sorting and release of CHIKV at the apical domain of polarized cells. Disruption of host cell myosin II, microtubule and microfilament networks did not disrupt the polarized release of CHIKV. However, treatment with tunicamycin resulted in a bi-directional release of CHIKV, suggesting that N-glycans of CHIKV envelope glycoproteins could serve as apical sorting signals.

Author Summary

Polarized cells are found in many parts of the human body and are characterized by the presence of two distinct plasma membrane domains: the apical domain facing the lumen and the basolateral domain facing the underlying tissues. Polarized epithelial cells line the major cavities of our body, while polarized endothelial cells line the blood-tissue interface, both of which protect our body against the invasion of biological pathogens. Thus, many pathogens have to invade the monolayer of epithelial or endothelial cells in order to establish infection. During infection with Chikungunya virus, a mosquito vector bites a human host and inoculates the virus into the host's bloodstream. In recent epidemics of Chikungunya infection, more severe clinical manifestations such as neurological complications were observed. As such, we studied the infection of Chikungunya virus in polarized cells in an aim to provide explanations for the more severe pathogenesis observed.