Naked DNA Immunization for Prevention of Prostate Cancer in a Dunning Rat Prostate Tumor Model
Abstract
We cloned into a pVax expression vector the secreted and the truncated (no peptide leader sequence) versions of the human prostate acid phosphatase (H-PAP-T), the human prostate- specific antigen (H-PSA-T) and the rat analogue of the human PSMA (R-"PSMA"-S). A stable transfectant with H-PSMA and H-PSA of the AT3B1 cell line was obtained. The following plasmids were produced and purified under GLP-conditions using the Qiagen technology: H-PSMA-T, R-"PSMA"-T, H-PSA, H-PSA-T, H-PAP-T and R"PSMA"-S. Preliminary studies using the Copenhagen rat tumor prostate model showed uniform tumor development in rats that were injected subcutaneously with 100 000 AT3B-lPSMA,PSA cells. Using a commercially available transfection device from Amaxa, we could transfect differentiated dendritic cells with 20-40% efficiency. Such transfected dendritic cells stimulated in vitro autologous T cells to PSMA. T cells cytotoxicity was then tested against tumor cells or peptide-pulsed T2 target cells. Both H-PSMA-T DCs and S-PSMA DCs generated antigen-specific cytotoxic T cell responses. The immune response was restricted towards one of four PSMA derived epitopes when priming and boosting was performed with S-PSMA. In contrast, T-PSMA transfected DCs primed T cells towards several PSMA derived epitopes. Subsequent repeated boosting with transfected DCs restricted the immune response to a single immunodominant epitope.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 2003
- Accession Number
- ADA417656
Entities
People
- Milcho S. Mincheff
Organizations
- George Washington University