A MAGE-A1 HLA-A A*0201 epitope identified by mass spectrometry.
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-A1(278-286) (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.
Pascolo
S
S
Institut for Cell Biology, Department Immunology, University of Tübingen, Auf der Morgenstelle 15, 72076 Tübingen, Germany. steve.pascolo@uni-tuebingen.de
Schirle
M
M
Gückel
B
B
Dumrese
T
T
Stumm
S
S
Kayser
S
S
Moris
A
A
Wallwiener
D
D
Rammensee
H G
HG
Stevanovic
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D001943:Breast Neoplasms / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000276:immunology; D015789:HLA-A2 Antigen / Q000276:immunology*; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D058950:Melanoma-Specific Antigens; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D010449:Peptide Mapping; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D010446:Peptide Fragments