A majority of cervical carcinomas contain human papillomavirus (HPV) types 16 or 18 (approximately 70%). The oncogenic association of these viruses is reflected in vitro in their ability to immortalize primary human keratinocytes. Expression of HPV E6 and E7 early genes is required for immortalization. These genes are invariably expressed in genital cancers and cervical carcinoma-derived cell lines. The biological and biochemical properties of these HPV early genes strongly suggest that they are the major factors in HPV associated oncogenesis. Lack of a tissue culture system for these viruses and their strict species and cell type tropism has hampered development of a model system for detailed analysis of HPV- induced tumorigenesis and the immune response during that process. Web propose to determine the feasibility of using genetic vaccination for the study of HPV-induced tumorigenesis and viral oncogene-specific immune responses. This method is based on the in vivo bombardment of tissues with DNA coated microprojectiles. This approach allows us to directly introduce foreign genes into cells of a living host in a fashion similar to what takes place in viral infections. We will target the natural host cells of HPV, keratinocytes, by calibration of the instrument and more importantly by using keratinocyte-specific promoters. We will employ keratin promoters shown previously to mediate foreign gene expression in keratinocytes. These studies will be carried out in mice because these animals have served as useful models of chemical-induced skin carcinogenesis; their immune system is well studied and many reagents are available for dissection of specific immune responses. By directly introducing foreign genes into mice, we hope to characterize HPV-mediated tumorigenesis; this system may provide a model for study of other intracellular pathogen-induced skin diseases.
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