期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2014
卷号：111
期号：46
页码：16544-16549
DOI：10.1073/pnas.1400818111
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceEpstein-Barr virus establishes a life-long latent infection (i.e., no virus is produced) in B cells in most people worldwide. A specific group of latency-associated proteins is expressed in B-cell and epithelial malignancies and likely contributes to tumorigenesis, but the role of epithelial cells in the virus life cycle is not well understood. We grew epithelial cells in organotypic cultures, allowing the cells to differentiate and stratify as they do in vivo. Unlike B-cell infection, EBV infection of epithelial cultures resulted in spontaneous production of high levels of virus, likely expanding the virus pool and increasing efficiency of transmission. This model of EBV-infected epithelium will enhance our understanding of the role of epithelial cells in the EBV life cycle. Epstein-Barr virus is a ubiquitous human herpesvirus associated with epithelial and lymphoid tumors. EBV is transmitted between human hosts in saliva and must cross the oral mucosal epithelium before infecting B lymphocytes, where it establishes a life-long infection. The latter process is well understood because it can be studied in vitro, but our knowledge of infection of epithelial cells has been limited by the inability to infect epithelial cells readily in vitro or to generate cell lines from EBV-infected epithelial tumors. Because epithelium exists as a stratified tissue in vivo, organotypic cultures may serve as a better model of EBV in epithelium than monolayer cultures. Here, we demonstrate that EBV is able to infect organotypic cultures of epithelial cells to establish a predominantly productive infection in the suprabasal layers of stratified epithelium, similar to that seen with Kaposi's-associated herpesvirus. These cells did express latency-associated proteins in addition to productive-cycle proteins, but a population of cells that exclusively expressed latency-associated viral proteins could not be detected; however, an inability to infect the basal layer would be unlike other herpesviruses examined in organotypic cultures. Furthermore, infection did not induce cellular proliferation, as it does in B cells, but instead resulted in cytopathic effects more commonly associated with productive viral replication. These data suggest that infection of epithelial cells is an integral part of viral spread, which typically does not result in the immortalization or enhanced growth of infected epithelial cells but rather in efficient production of virus.
