Characterization of New Breast Tumor-Specific Antigens Using a Novel Antigen Discovery System
Final rept. 1 Jun 1997-31 May 2000
PITTSBURGH UNIV PA
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The goal of this project is to devise a new method for antigen discovery using dendritic cells as antigen-presenting cells to prime autologous naive CD4 and CD8 T cells from healthy donors against breast tumor proteins and peptides. To identify HLA Class I-restricted tumor antigens, we extracted peptides from tumor HLA Class I molecules, fractionated them by RP-HPLC, and loaded them onto DCs to prime naive CD8 T cells in vitro. Our results show that we were able to prime naive CD8 T cells to several peptide fractions and generate specificity to the tumor. Electrospray ionization mass spectrometry of the immunostimulatory peptide fractions identified a panel of peptides that could bind MHC-Class I molecules. To identify HLA Class II-restricted tumor antigens, we fractionated tumor protein extracts using RP-HPLC, and loaded them onto DCs to prime naive CD4 T cells in vitro. Our results show that we were able to prime naive CD4 T cells to several protein fractions and generate specificity to the tumor. In summary, results stemming from this project show that we have created a novel method that identified a panel of potentially new MHC Class I- and Class II- restricted tumor antigens.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research