Identification of H-2Kb binding and immunogenic peptides from human papilloma virus tumour antigens E6 and E7

Abstract

Peptides can be used to induce MHC class I restricted cytotoxic T cells (CTL) through in vivo immunization. This approach may enable the development of peptide vaccination schemes for immunization against viral infection in humans. Human papillomavirus (HPV) is one of a few viruses associated with human cancer and the development of an anti-cancer vaccine seems possible. As a model approach, we searched the E6 and E7 proteins of the human papillomavirus type 16 for possible murine MHC class I restricted peptide epitopes. We utilized the mouse H2-Kb peptide binding motif which consists of phenylalanine or tyrosine at position five and leucine at the carboxy-terminus with the modification that leucine could be replaced by other aliphatic but non-aromatic amino acids. Four peptide sequences from E6 and two from E7 were selected. These peptides were tested for their ability to bind and stabilize Kb and for their immunogenicity in vivo. It was shown that one peptide from E6, E6.1 (50-57), bound Kb, but was not able to prime mice in vivo. In contrast, the two selected E7 peptides E7.1 (21-28) and E7.2 (48-55) bound Kb and were immunogenic in vivo. The peptide induced CTL lysed syngeneic EL-4 cells transfected with the open reading frame of E7 but not vector only transfectants. This implies that both peptides were naturally processed and presented by Kb on the surface of target cells. MHC class I peptide binding motifs therefore appear to be an effective and useful tool to predict peptide epitopes of proteins associated with cancer.