Autologous Human Monocyte-Derived Dendritic Cells Genetically Modified to Express Melanoma Antigens Elicit Primary Cytotoxic T Cell Responses In Vitro: Enhancement by Cotransfection of Genes Encoding the Th1-Biasing Cytokines IL-12 and IFN-α

摘要: DNA-based immunization strategies designed to elicit cellular antitumor immunity offer an attractive alternative protein- or peptide-based approaches. In the present study we have evaluated feasibility of DNA vaccination for induction CTL reactivity five different melanoma Ags in vitro. Cultured, monocyte-derived dendritic cells (DC) were transiently transfected with plasmid encoding human MART-1/Melan-A, pMel-17/gp100, tyrosinase, MAGE-1, MAGE-3 by particle bombardment and used stimulate autologous PBMC responder T cells. these previously identified was reproducibly generated after two three stimulations genetically modified DC. Co-ordinate transfection Ag cDNAs into DC promoted responders capable recognizing epitopes from both gene products. Coinsertion genes Th1-biasing cytokines IL-12 IFN-alpha consistently enhanced magnitude resulting Ag-specific reactivity. Importantly, a single cDNA stimulating restricted multiple host MHC alleles, some which had not been identified. These results support inherent strengths gene-based vaccine approaches that do require prior knowledge haplotypes relevant MHC-restricted peptide epitopes. Given previous observations situ tumor HLA allele-loss variants, may greater diversity therapeutic immunity, thereby enhancing clinical utility success such