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Divisions of
*
Surgical Oncology,
Hematology/Oncology, and
Experimental Radiation Oncology, University of California Los Angeles Medical Center, Los Angeles, CA 90095, and
§
Department of Medicine, University of Connecticut Health Center, Farmington, CT 06032
Dendritic cells (DC) are potent stimulators of primary T cell
responses. In this study, we demonstrate that DC, genetically
engineered to express the MART-1/Melan-A (MART-1) tumor-associated Ag,
express MART-1 mRNA and protein, correctly process and present the
HLA-A2.1-restricted immunodominant MART-1 peptide
(MART-127–35), and serve as potent stimulators of
MART-1-specific CTL in vitro. A replication-defective E1-deleted
adenovirus (AdV) was constructed that expresses MART-1 (AdVMART1).
Transduced DC produce full length MART-1 mRNA as well as MART-1
protein. AdVMART1 does not significantly down-regulate cell surface
class I expression despite having an intact E3 region. Transduction of
an HLA-A2-positive/MART-1-negative cell line with AdVMART1 renders
these cells sensitive to lysis by CTL specific for the
MART-127–35 immunodominant peptide. In addition, DC
transduced with AdVMART1 stimulated MART-127–35-specific
tumor-infiltrating lymphocytes to synthesize IFN-
. Finally,
AdVMART1-transduced DC were able to generate MART-127–35
peptide-specific, class I-restricted CTL in PBL cultures from normal
donors. This study supports the use of tumor Ag-engineered DC in
genetic immunotherapy.
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