Immunization against human papillomavirus infection and associated neoplasia

Chimeric virus like particles (CVLPs) constructed by fusing human papillomavirus type 16 (HPV16) E7 sequences into the C-terminus of the viral L1 gene constitute the first generation of preventive and therapeutic HPV vaccines. Even though vaccination with DNA is highly efficient in the induction of a cytotoxic T-cell (CTL) response utilization of a DNA vaccine in the HPV context, it has been hampered by concern for the oncogenic potential of the E6 and E7 proteins encoded by the viral oncogenes.

To consider the use and impact of E7 DNA for immunization.

In addition to hemagglutination inhibition, a versatile assay to measure neutralization of yeast cell-derived pseudovirions carrying a green fluorescence reporter gene has now been developed. Mice immunized with the HPV16 CVLPs generate E7-specific CTLs, which kill E7 expressing or E7 peptide loaded RMA-cells, protect against tumor formation by syngeneic HPV transformed cells and also induce regression of already established tumors. Since generation of CTL response is achieved by presentation of epitopes as short peptides together with appropriate MHC class I molecules, complete proteins are not required. Instead a shuffled E7 protein has now been used successfully for generating CTL responses comparable to the CVLP responses in mice.

Our preliminary results suggest that immunization with E7 shuffled DNA yields a response directed against the authentic E7 protein. Furthermore, booster immunization with E7 shuffled DNA would avoid inhibition by neutralizing antibodies, however, further studies are needed to guarantee that the shuffled E7 protein lacks oncogenic activity.