Cytokine-FC fusion genes as molecular adjuvants for DNA vaccines.
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.
Hirschhorn-Cymerman
Daniel
D
Memorial Sloan-Kettering Cancer Center and Weill Medical College of Cornell University, New York, NY, USA.
Perales
Miguel-Angel
MA
D000276:Adjuvants, Immunologic / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000627:therapeutic use; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics* / Q000627:therapeutic use; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000627:therapeutic use*; D014162:Transfection; D019444:Vaccines, DNA / Q000235:genetics / Q000627:therapeutic use*
D000276:Adjuvants, Immunologic; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D016178:Granulocyte-Macrophage Colony-Stimulating Factor