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[Cancer Research 61, 4072-4077, May 15, 2001]
© 2001 American Association for Cancer Research

Immunology

A MAGE-A1 HLA-A*0201 Epitope Identified by Mass Spectrometry1

Steve Pascolo2,,3, Markus Schirle2, Brigitte Gückel, Tilman Dumrese4, Susanne Stumm, Simone Kayser, Arnaud Moris, Diethelm Wallwiener, Hans-Georg Rammensee and Stefan Stevanovic

Institut for Cell Biology, Department Immunology [S. P., M. S., A. M., H-G. R., S. Ste.], and Department of Obstetrics and Gynecology, University of Tübingen [B. G., S. Stu., S. K., D. W.], 72076 Tübingen, Germany

Peptides presented by HLA-A*0201 molecules on the surface of the human breast carcinoma cell line KS24.22 after IFN-{gamma} induction were analyzed by the "Predict-Calibrate-Detect" approach, which combines epitope prediction and high-performance liquid chromatography mass spectrometry. One of the predicted epitopes, MAGE-A1278–286 (KVLEYVIKV), was found to be presented by HLA-A*0201, with an estimated copy number of 18 molecules/cell. HLA-A*0201 transgenic mice (HHD mice) were used to generate CTL lines that stained positive with an HLA-A*0201 tetramer folded around the KVLEYVIKV peptide and killed peptide-loaded mouse target cells expressing HLA-A*0201. IFN-{gamma}-treated or -nontreated HLA-A*0201 expressing HeLa cells transiently transfected with a plasmid expressing the MAGE-A1 gene stimulated in vitro cytokine production by the CTL lines. Moreover, IFN-{gamma}-treated KS24.22 cells, but not IFN-{gamma}-treated HLA-A*0201+ MAGE-A1- cells or IFN-{gamma}-treated HLA-A*0201- MAGE-A1+ cells, were killed by these CTLs. Thus, the combination of HLA epitope prediction, peptide analysis, and immunological methods is a powerful approach for the identification of tumor-associated epitopes.




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Copyright © 2001 by the American Association for Cancer Research.