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-alpha.
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-alpha 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.
Tüting
T
T
Department of Surgery, University of Pittsburgh School of Medicine, University of Pittsburgh Cancer Institute, PA 15261, USA.
Wilson
C C
CC
Martin
D M
DM
Kasamon
Y L
YL
Rowles
J
J
Ma
D I
DI
Slingluff
C L
CL
Wagner
S N
SN
van der Bruggen
P
P
Baar
J
J
Lotze
M T
MT
Storkus
W J
WJ
D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics*; D018664:Interleukin-12 / Q000096:biosynthesis / Q000235:genetics*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D058965:MART-1 Antigen; D008545:Melanoma / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D009000:Monocytes / Q000276:immunology; D016254:Mutagenesis, Insertional / Q000276:immunology; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology; D010455:Peptides / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D018417:Th1 Cells / Q000378:metabolism*; D014162:Transfection / Q000276:immunology*; D014407:Tumor Cells, Cultured; D019444:Vaccines, DNA / Q000276:immunology; D058951:gp100 Melanoma Antigen
D000951:Antigens, Neoplasm; D018076:DNA, Complementary; D018984:Epitopes, T-Lymphocyte; C072013:MAGEA3 protein, human; D058965:MART-1 Antigen; C548667:MLANA protein, human; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C548637:PMEL protein, human; D010455:Peptides; D019444:Vaccines, DNA; D058951:gp100 Melanoma Antigen; D018664:Interleukin-12; D007371:Interferon-gamma; D008156:Luciferases