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Journal of Virology, June 2003, p. 6533-6540, Vol. 77, No. 11
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.11.6533-6540.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Activation of Adenovirus Early Promoters and Lytic Phase in Differentiated Strata of Organotypic Cultures of Human Keratinocytes

Francisco Noya,1 Cristina Balagué,2,{dagger} N. Sanjib Banerjee,1 David T. Curiel,2,3 Thomas R. Broker,1 and Louise T. Chow1*

Department of Biochemistry and Molecular Genetics,1 Departments of Medicine, Pathology, and Surgery, Division of Human Gene Therapy,3 Gene Therapy Center, University of Alabama at Birmingham, Birmingham, Alabama 35294-00052

Received 27 November 2002/ Accepted 10 March 2003

Human oncolytic adenoviruses have been used in clinical trials targeting cancers of epithelial origin. To gain a better understanding of the infectious cycle of adenovirus in normal human squamous tissues, we examined the viral infection process in organotypic cultures of primary human keratinocytes. We show that for the infection to occur, wounding of the epithelium is required. In addition, infection appears to initiate at the basal or parabasal cells that express the high-affinity coxsackievirus-adenovirus receptor, CAR, whereas the productive phase takes place in differentiated cells. This is due, at least in part, to the differentiation-dependent activation of the E1A and E2A early promoters and E4 promoters. We also show that adenovirus infection triggers a response mediated by the abnormal accumulation of cyclin E and p21cip1 proteins similar to the one previously observed in human papillomavirus-infected tissues. However, the virus seems to be able to overcome it, at least partially.


* Corresponding author. Mailing address: Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, 510 McCallum Basic Health Sciences Bldg., 1918 University Blvd., Birmingham, AL 35294-0005. Phone: (205) 975-8300. Fax: (205) 975-6075. E-mail: ltchow{at}uab.edu.

{dagger} Present address: Almirall Prodesfarma, Target Validation Unit, Barcelona Science Park, Barcelona 08028, Spain.


Journal of Virology, June 2003, p. 6533-6540, Vol. 77, No. 11
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.11.6533-6540.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




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