Immunogenicity of recombinant chaperone-complexed antigens

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We will study the immunogenicity of antigen/hsp complexes that are either expressed (as endogenous complexes) by antigen-presenting cells (APC), or taken up by APC as exogenous complexes, processed and presented to lymphocytes. Methods for the production, purification and characterization of recombinant hsp73- fusion antigen complexes will be refined. The cell biology of processing exogenous or endogenous hsp/antigen complexes for MHC class I-restricted epitope presentation to cytotoxic T lymphocytes (CTL) will be studied. The main focus will be on the intracellular traffic of exogenous or endogenous hsp/antigen complexes, the subcellular site at which processing takes place, the site and kinetic of the generation of presentation-competent MHC class I molecules, and the proteolytic system(s) involved. Attempts will be made to facilitate proteolytic processing of hsp-bound recombinant fusion antigen by flanking the relevant epitopes with cathepsin-sensitive cleavage sites. These in vitro studies will be complemented by in vivo studies that will elucidate priming of humoral and cellular immune responses in vivo by DNA- or protein-based vaccines containing hsp/antigen complexes. The project makes an effort to integrate the in vitro study of the cell biology of processing and presentation with the in vivo study of the immunogenicity of hsp/antigen complexes that are a fascinating, novel approach of T cell-stimulating vaccines based on a natural adjuvant.