Human papillomaviruses (HPV) are associated with malignant cervical neoplasia. Several HPV-related diseases have been shown to be sensitive to interferon (IFN) treatment. HeLa cells contain and express the HPV type 18 genome and were used as a model for the evaluation of the viral expression regulation and the effect on the malignant phenotype during IFN treatment. Cells were treated continuously with 200 IU/ml IFN-alpha 2b or natural leukocyte INF-alpha for six passages (42 days). Some IFN-induced changes were observed: decrease of HPV-18 mRNA expression, changes of cell morphology, and reduction of clonogenicity in soft agar. Tumorigenicity in nude mice was not modified. Other targets of the IFN system were analyzed, and an increase of the 2',5'-oligoadenylate synthetase mRNA level and a down-regulation of type I IFN receptor were found. These results demonstrate that long-term IFN-alpha treatment induces a partial phenotypic reversion of HeLa cells to a more differentiated stage were down-regulation of HPV-18 expression could play a central role. It therefore confirms that the IFN-alpha treatment may be therapeutically useful in cervix cancer produced by HPV-18.