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Journal of Virology, April 2004, p. 4074-4084, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.4074-4084.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Wild-Type Kaposi's Sarcoma-Associated Herpesvirus Isolated from the Oropharynx of Immune-Competent Individuals Has Tropism for Cultured Oral Epithelial Cells

Karen M. Duus,1 Vivian Lentchitsky,2 Timothy Wagenaar,3 Charles Grose,3 and Jennifer Webster-Cyriaque1,2,4*

Lineberger Comprehensive Cancer Center,1 Department of Dental Ecology, University of North Carolina Dental School,4 Department of Microbiology and Immunology, University of North Carolina Medical School, University of North Carolina, Chapel Hill, North Carolina 27599,2 Department of Pediatrics and Central Microscopy Research Facility, University of Iowa, Iowa City, Iowa 522423

Received 28 July 2003/ Accepted 4 December 2003

Based on the observation that wild-type Kaposi's sarcoma-associated herpesvirus (KSHV) DNA can be detected in the oral cavity of healthy, immunocompetent individuals, we hypothesized that epithelial cells could be infected in vitro by wild-type (WT) KSHV isolated from immunocompetent individuals. Primary oral epithelial (P-EPI) cells and telomerase-immortalized oral epithelial cells were generated from human gingival tissue and were then infected in vitro with WT KSHV isolated from throat wash samples. Markers of lytic and latent KSHV infection were detected in cultures by 24 h postinfection by immunofluorescence confocal microscopic assays. The infectivity of the WT and BCBL virus was blocked by neutralizing antibodies against KSHV gB. The presence of KSHV DNA in these cells was confirmed by real-time PCR amplification of different regions of the viral genome. The significant in vitro viral replication that had occurred was inhibited by ganciclovir and by neutralizing antibodies against gB. When infected cultures were examined by scanning electron microscopy, thousands of KSHV particles were clearly visible across the surfaces of P-EPI cells. The detection of enveloped particles indicated that the infectious cycle had proceeded through assembly and egress. We thus demonstrated that oral WT KSHV isolated from immunocompetent individuals was able to infect and replicate in vitro in a relevant primary cell type. Furthermore, our results provide compelling evidence for KSHV transmission within infected oral epithelial cells derived from healthy, immunocompetent populations.

* Corresponding author. Mailing address: University of North Carolina, Room 113 Dental Research Center, Campus Box 7455, Chapel Hill, NC 27599. Phone: (919) 943-6198. Fax: (919) 966-3683. E-mail: jennifer_webster-cyriaque{at}med.unc.edu.

Journal of Virology, April 2004, p. 4074-4084, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.4074-4084.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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