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Journal of Virology, May 2002, p. 4723-4733, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.4723-4733.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Infectious Virions Produced from a Human Papillomavirus Type 18/16 Genomic DNA Chimera

Craig Meyers,1* Jennifer L. Bromberg-White,1 Jiaping Zhang,2 Michelle E. Kaupas,2 Janine T. Bryan,2 Robert S. Lowe,2 and Kathrin U. Jansen2

Department of Microbiology and Immunology, College of Medicine, The Pennsylvania State University, Hershey, Pennsylvania 17033,1 Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania 194862

Received 20 July 2001/ Accepted 13 December 2001

The organotypic raft culture system has allowed the study of the differentiation-dependent aspects of the human papillomavirus (HPV) life cycle. However, genetic strategies to more completely understand the HPV life cycle are limited. The generation of chimeric viruses has been a useful tool in other virus systems to analyze infection and replication. To investigate the specificity of the interaction of nonstructural genes of one HPV type with the structural genes of another HPV type, we have replaced the L2 and L1 open reading frames (ORFs) of HPV type 18 (HPV18) with the L2 and L1 ORFs of HPV type 16 (HPV16). The resulting HPV18/16 chimeric construct was introduced into primary keratinocytes, where it was stably maintained episomally at a range of 50 to 100 copies of HPV genomic DNA, similar to that typically found in HPV-infected cells in vivo. The integrity of the HPV18/16 genomic DNA chimera was demonstrated. Upon differentiation in raft cultures, late viral functions, including viral DNA amplification, capsid gene expression, and virion morphogenesis, occurred. Chimeric HPV18/16 virions were purified from the raft cultures and were capable of infecting keratinocytes in vitro. Additionally, infection was specifically neutralized with human HPV16 virus-like particle (VLP)-specific antiserum and not with human HPV18 VLP-specific antiserum. Our data demonstrate that the nonstructural genes of HPV18 functionally interact with the structural genes of HPV16, allowing the complete HPV life cycle to occur. We believe that this is the first report of the propagation of chimeric HPV by normal life cycle pathways.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, College of Medicine, The Pennsylvania State University, Hershey, PA 17033. Phone: (717) 531-6240. Fax: (717) 531-4600. E-mail: cmm10{at}psu.edu.

Journal of Virology, May 2002, p. 4723-4733, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.4723-4733.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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