title: Efficient replication of Epstein-Barr virus in stratified epithelium in vitro.
abstract: 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.
authors.affiliation: Departments of Microbiology and Immunology.
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Temple
authors.affiliation: Public Health Sciences, and The Penn State Hershey Cancer Institute, Hershey, PA 17033.
authors.forename: Junjia
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Pathology, The Pennsylvania State University College of Medicine, Hershey, PA 17033; and.
authors.forename: Lynn
authors.initials: L
authors.lastname: Budgeon
authors.affiliation: Departments of Microbiology and Immunology, The Penn State Hershey Cancer Institute, Hershey, PA 17033 Pathology, The Pennsylvania State University College of Medicine, Hershey, PA 17033; and.
authors.forename: Neil David
authors.initials: ND
authors.lastname: Christensen
authors.affiliation: Departments of Microbiology and Immunology, The Penn State Hershey Cancer Institute, Hershey, PA 17033.
authors.forename: Craig
authors.initials: C
authors.lastname: Meyers
authors.affiliation: Departments of Microbiology and Immunology, The Penn State Hershey Cancer Institute, Hershey, PA 17033 csample@hmc.psu.edu.
authors.forename: Clare E
authors.initials: CE
authors.lastname: Sample
mesh_terms: D000212:Acyclovir / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003588:Cytopathogenic Effect, Viral; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D019309:Epstein-Barr Virus Nuclear Antigens / Q000096:biosynthesis / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D005881:Gingiva / Q000166:cytology; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology*; D007633:Keratins / Q000032:analysis; D014066:Palatine Tonsil / Q000166:cytology; D010957:Plasmids / Q000235:genetics; D011498:Protein Precursors / Q000032:analysis; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014776:Virus Cultivation; D017735:Virus Latency; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; C063676:BZLF1 protein, Herpesvirus 4, Human; D004279:DNA, Viral; C402641:EBNA-2 protein, Human herpesvirus 4; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; C099537:Epstein-Barr virus encoded RNA 1; D011498:Protein Precursors; D012367:RNA, Viral; D015534:Trans-Activators; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C032253:involucrin; D007633:Keratins; C417029:EBV-encoded nuclear antigen 1; D000212:Acyclovir
keywords: Epstein–Barr virus; epithelial; organotypic culture; productive replication