HUMAN-PAPILLOMAVIRUS TYPE-16 E7 PROTEIN SENSITIZES CERVICAL KERATINOCYTES TO APOPTOSIS AND RELEASE OF INTERLEUKIN-1-ALPHA

Interleukin-1 alpha. (IL-1 alpha) is a multifunctional cytokine that p romotes inflammation, tissue remodeling and epithelial hyperplasia, Ke ratinocytes produce and sequester large amounts of biologically active IL-1 alpha which can be released after injury or infection. We show t hat high level expression of human papillomavirus (HPV) type 16 E6 and E7 oncoproteins enhanced release of IL-1 alpha from cultures of norma l cervical keratinocytes (relative effectiveness E7>E6/E7 much greater than E6> control). The amount of IL-1 alpha. released was directly re lated to the ability of E7 or E6/E7 to stimulate apoptosis, E7 protein s that bound the retinoblastoma protein (Rb) strongly (HPV-16 and -18) induced more IL-1 alpha release than those that bound poorly (HPV-6 a nd an HPV-16 E7 24gly mutant). Furthermore, overexpression of the E2F- 1 transcription factor, a downstream target of Rb, induced extensive a poptosis and IL-1 alpha release. Apoptosis and IL-1 alpha release in r esponse to growth factor removal occurred in part through a p53-indepe ndent pathway as coexpression of E6 and downregulation of p53 did not prevent either response. Immunohistochemical analyses showed that IL-1 alpha. was expressed by keratinocytes in normal cervical epithelia, l ow and high grade dysplasias, and cervical carcinomas. However, HPV-16 E6/E7 RNA expression and apoptosis increased in parallel in prolifera ting keratinocytes in severe dysplasias and carcinomas suggesting that IL-1 alpha release is associated with progression to high grade disea se. Thus, high level expression of the HPV-16 E7 protein sensitizes ke ratinocytes to apoptosis which results in release of IL-1 alpha.