The major objective of this proposal is to develop a new generation of more rationally designed vaccines against viruses and (virus-induced) cancer. The major vaccine targets are cytomegalovirus (CMV) and p53. Additional targets are human papillomavirus type 16 (HPV16) and hepatitis C virus (HCV). To achieve this overall objective a collaboration has been set up between 6 academic and research institutes and two European Vaccine companies (Pasteur-Mérieux and Biocine). In a multi-disciplinary approach partners 1,2 and 4-6 will collaborate to delineate the detailed rules for processing and presentation of MHC class I binding peptides.
Subsequent steps analysed are 1) characterization of proteasome cleavage sites and specificity 2) mechanism and specificity of TAP-mediated transport from cytosol into endoplasmic reticulum 3) identification of best immunogenic peptides by MHC class I and class ll binding and stability assays followed by in-vitro response induction assays with human responding T-cells and immunization of HLA transgenic mice. Together the data obtained will be utilised to predict optimal MHC class I epitopes. A separate analysis, conducted in the laboratory of the coordinator, will yield information on optimal MHC class ll epitopes. This information will be used as building blocks of new vaccines and to monitor response against vaccines. Intact antigen-based vaccines and string of bead epitope-based vaccines will be tested in virus and tumor challenge/protection experiments in normal and HLA-transgenic animals in which antibody, T helper and CTL responses will be monitored. Various models of antigen delivery in the vaccines will be compared with respect to efficiency of protection and efficiency of T-cell response induction, including peptides or protein in adjuvant, peptides or protein loaded onto dendritic cells, accompanied by in-vitro analysis of optimal loading of dendritic cells with protein for induction of T helper and CTL responses (partner 1), and DNA as well as canary pox virus vector vaccines (both provided by partner 3). Partner 7 has an important role in defining the influence of tissue-specific promotors and secretory signals on the effectiveness of DNA vaccination and this knowledge will be used for optimal DNA and viral vector vaccine design by partner 3. Partner 8 will be involved in the study of antigen processing by DC, using retroviral vector introduction of antigen into DC as an alternative to protein loading of DC. The combined efforts of these laboratories with highly complementary expertise are expected to allow the construction of a new generation of effective and safe vaccines for a number of viral diseases and specific cancers associated with p53 overexpression or HPV16.
Funding SchemeCSC - Cost-sharing contracts
RG20 7NN Newbury
1066 CX Amsterdam