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1
,
Departments of
*
Surgery,
Molecular Genetics and Biochemistry,
Pathology, and
§
Internal Medicine, University of Pittsburgh School of Medicine, and University of Pittsburgh Cancer Institute, Pittsburgh, PA 15261;
¶
Department of Surgery, University of Virginia, Charlottesville, VA 22908;
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Department of Dermatology, University of Essen School of Medicine, Essen, Germany; and
#
Ludwig Institute for Cancer Research, Brussels Branch, Brussels, Belgium
DNA-based immunization strategies designed to elicit cellular
antitumor immunity offer an attractive alternative to protein- or
peptide-based approaches. In the present study we have evaluated the
feasibility of DNA vaccination for the induction of CTL reactivity to
five different melanoma Ags in vitro. Cultured, monocyte-derived
dendritic cells (DC) were transiently transfected with plasmid DNA
encoding human MART-1/Melan-A, pMel-17/gp100, tyrosinase, MAGE-1, or
MAGE-3 by particle bombardment and used to stimulate autologous PBMC
responder T cells. CTL reactivity to these previously identified
melanoma Ags was reproducibly generated after two or three stimulations
with genetically modified DC. Co-ordinate transfection of two melanoma
Ag cDNAs into DC promoted CTL responders capable of recognizing
epitopes from both gene products. Coinsertion of genes encoding the
Th1-biasing cytokines IL-12 or IFN-
consistently enhanced the
magnitude of the resulting Ag-specific CTL reactivity. Importantly, DC
transfected with a single melanoma Ag cDNA were capable of stimulating
Ag-specific CTL reactivity restricted by multiple host MHC alleles,
some of which had not been previously identified. These results support
the inherent strengths of gene-based vaccine approaches that do not
require prior knowledge of responder MHC haplotypes or of relevant
MHC-restricted peptide epitopes. Given previous observations of in situ
tumor HLA allele-loss variants, DC gene vaccine strategies may elicit a
greater diversity of host therapeutic immunity, thereby enhancing the
clinical utility and success of such approaches.
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