Air-Liquid Interface Method To Study Epstein-Barr Virus Pathogenesis in Nasopharyngeal Epithelial Cells

Abstract

Lifting adherent cells to the air-liquid interface (ALI) is a method conventionally used to culture airway epithelial cells into polarized apical and basolateral surfaces. Reactivation of Epstein-Barr virus (EBV) from monolayer epithelial cultures is sometimes abortive, which may be attributed to the lack of authentic reactivation triggers that occur in stratified epithelium in vivo. In the present work, the ALI culture method was applied to study EBV reactivation in nasopharyngeal epithelial cells. The ALI culture of an EBV-infected cell line yielded high titers and can be dissected by a variety of molecular virology assays that measure induction of the EBV lytic cascade and EBV genome replication and assembly. EBV infection of polarized cultures of primary epithelial cells can be challenging and can have variable efficiencies. However, the use of the ALI method with established EBV-infected cell lines offers a readily available and reproducible approach for the study of EBV permissive replication in polarized epithelia.

Circular DNA tumor viruses make circular RNAs

“…The linear limits of detection were between 4 and 4 × 10 8 copies per reaction. Reactions were assembled as previously described ( 62 ). Input genomic DNA was normalized and compared with a reference cell line (Raji) averaging 50 EBV episomal copies per cell.…”

Air-Liquid Interface System To Understand Epstein-Barr Virus-Associated Nasopharyngeal Carcinoma

“…As the organotypic raft culture system is not widely used, Caves et al explored the use of a transwell-based ALI culture system to probe EBV replication in differentiated epithelial cells ( 10 ). The ALI system is conventionally used to polarize primary airway epithelial cells.…”

Epstein–Barr Virus Infection of Pseudostratified Nasopharyngeal Epithelium Disrupts Epithelial Integrity

“…A variety of in vitro epithelial cell models for EBV infection have been developed, ranging from monolayer unpolarized epithelial cells to polarized epithelial cells, using oropharyngeal or tonsillar epithelial cells [ 4 , 5 , 6 , 7 , 8 ]. Although many useful data have been generated using these models, they may not closely represent the physiological condition in the nasopharynx, for the following reasons.…”