Cytokine-FC Fusion Genes as Molecular Adjuvants for DNA Vaccines

Abstract

The use of gene constructs for DNA immunization offers several potential advantages over other commonly used vaccine approaches: (1) full-length cDNA provides multiple potential class I and class II epitopes, thus bypassing limitations of MHC restriction; (2) bacterial plasmid DNA contains immunogenic unmethylated CpG motifs (immunostimulatory sequences) that may act as a potent immunological adjuvant; and (3) DNA is relatively simple to purify in large quantities. The cDNA encoding the antigen of interest is cloned into a bacterial expression plasmid with a constitutively active promoter and this plasmid is injected into the skin or muscle where it is taken up by professional antigen-presenting cells, particularly dendritic cells, either through direct transfection or cross-priming. One can further enhance or modulate the immune response through co-delivery of DNA encoding cytokines or chemokines, including cytokine-Fc fusion molecules. The latter use molecular techniques to fuse a cytokine to the Fc portion of IgG1, creating a chimeric molecule with functional activity. In the present chapter, we will outline the approach to develop cytokine-Fc fusion genes as molecular adjuvants and will use GM-CSF as an example.