Journal article
Propagation of infectious human papillomavirus type 16 by using an adenovirus and Cre/LoxP mechanism
Proceedings of the National Academy of Sciences - PNAS, Vol.101(7), pp.2094-2099
02/17/2004
DOI: 10.1073/pnas.0308615100
PMCID: PMC357057
PMID: 14769917
Abstract
Human papillomavirus type 16 (HPV16) infection is a major risk factor for the development of squamous cell cancers of the cervix and of the head and neck. A major barrier to understanding the progression from initial infection to cancer has been the lack of
in vitro
models that allow infection, replication, and persistence of the viral genome as an episome in differentiated epithelial cells. To overcome this barrier, we designed an adenoviral delivery vector that contained a full HPV16 genome flanked by LoxP homologous recombination sites and a fluorescent reporter that was expressed only after the HPV genome was excised by Cre recombinase. This system delivered circular HPV16 genomes to cervical epithelial cells and well differentiated human airway epithelia. After delivery, the HPV16 genome replicated and persisted as an episome in cervical keratinocytes. These cells developed an immortalized phenotype and a dysplastic epithelial appearance. Moreover, induction of differentiation led to the expression of late genes and production of infectious HPV16 virions. This work provides a means of introducing biologically active HPV genomes into epithelial cells, which are normally difficult to transfect. These methods allow the study of HPV genome replication and gene expression in the earliest stages of HPV genome establishment, and they may provide a means to study nononcogenic HPV viral types.
Details
- Title: Subtitle
- Propagation of infectious human papillomavirus type 16 by using an adenovirus and Cre/LoxP mechanism
- Creators
- John H Lee - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131Su Min P Yi - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131Mary E Anderson - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131Kristi L Berger - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131Michael J Welsh - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131Aloysius J Klingelhutz - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131Michelle A Ozbun - Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.101(7), pp.2094-2099
- DOI
- 10.1073/pnas.0308615100
- PMID
- 14769917
- PMCID
- PMC357057
- NLM abbreviation
- Proc Natl Acad Sci U S A
- ISSN
- 0027-8424
- eISSN
- 1091-6490
- Publisher
- National Academy of Sciences
- Language
- English
- Date published
- 02/17/2004
- Academic Unit
- Neurology; Molecular Physiology and Biophysics; Microbiology and Immunology; Radiation Oncology; Neurosurgery; Internal Medicine
- Record Identifier
- 9984001135902771
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