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A new strategy for tumor antigen discovery based on in vitro priming of naïve T cells with dendritic cells
Henry Kao; Andrew A. Amoscato; Pawel Ciborowski; Olivera J. Finn (Profiled Author: Pawel S Ciborowski)
Clinical Cancer Research 2001;7(11 SUPPL.):773s-780s.Abstract
We describe a method for discovery of new tumor antigens that uses dendritic cells (DCs) as antigen-presenting cells to prime autologous naïve CD4 + and CD8 + T cells from healthy donors against tumor proteins and peptides. For the identification of HLA class I-restricted tumor antigens, peptides were extracted from tumor HLA class I molecules, fractionated by reverse phase-high performance liquid chromatography, and loaded onto in vitro-generated DCs to prime naïve CD8 + T cells. Our results show that we were able to prime naïve CD8 + T cells in vitro to several peptide fractions and generate specificity for the tumor. Electrospray ionization mass spectrometry was used to confirm that these fractions contained peptides derived from MHC class I molecules, and the primed CD8 + T cells were used to further analyze the immunostimulatory peptide fractions. For the identification of HLA class II-restricted tumor antigens, we fractionated tumor protein extracts using reverse phase-high performance liquid chromatography and loaded individual fractions onto DCs to prime naïve CD4 + T cells. Our results show that we were also able to prime naïve CD4 + T cells to several protein fractions and generate specificity for the tumor. These results illustrate the potential of this method to identify new immunostimulatory MHC class I- and class II-restricted tumor antigens.
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