Development of a Novel Marker Gene Based Assay System for Detection and Evaluation of Antiviral Agents with Activity against Papillomaviruses

Abstract

A novel assay system has been developed in which expression of a human tissue-plasminogen activator (t-PA) gene, carried on a recombinant papillomavirus vector, is used as a marker for the presence of bovine papillomavirus type 1 (BPV-1) within transformed mouse C127 cells. This provides a relatively quick and simple means of identifying and evaluating agents with anti-papillomavirus activity. Using this system the antiviral activity and cytotoxicity of interferon and retinoic acid, have been investigated. After seven subcultures in the presence of 200 Units ml−1mouse α and β interferon, t-PA expression was completely inhibited, with a concurrent alteration in cellular morphology, and restoration of contact inhibition. In accordance with the problems encountered with interferon therapy of human papillomavirus infections, these effects were dependent on the continued presence of interferon, its removal leading to a rapid return of t-PA expression, and reversion of cells to the transformed phenotype. In comparison, 2.0 μg ml−1retinoic acid partially reduced t-PA expression (this effect was largely maintained even after removal of the inhibiting compound) but did not affect the transformed cell phenotype. These results are discussed in relation to other in vitro studies and also to the clinical treatment of human papillomavirus (HPV) disease.