Isolation of novel HLA-DR restricted potential tumor-associated antigens from the melanoma cell line FM3.
Endogenous peptides bound to the constitutively expressed MHC class II molecules HLA-DR and HLA-DQ of the melanoma cell line FM3 were examined. By a combination of analytical methods (narrow bore and capillary reversed-phase high-performance liquid chromatography with subsequent spotting on polyvinylidene difluoride membranes, matrix-assisted laser desorption ionization mass spectrometry, and Edman microsequencing), we were able to isolate and identify a panel of HLA-DR4/2 (HLA-DRB1*0401/0201/DRB5*0101)-associated self-peptides from the melanoma cell line FM3. Among ubiquitously HLA-DR-associated peptides such as peptides from the class II-associated invariant chain peptide region of the invariant chain, HLA-class I, the transferrin receptor, and the IFN-gamma receptor, we identified several potential tumor-associated antigens stemming from the MHC class I-restricted tumor antigen gp100, the Ca2(+)-binding protein annexin II, and proteins from the hsp70 family. Chinese hamster ovary cells cotransfected with HLA-DRA, DRB1*0401, and CD80 genes were shown specifically to prime T lymphocytes from HLA-DRB1*0401 donors to the annexin II and gp100 peptides. These results demonstrate that MHC class II molecules expressed by melanoma cells potentially present a variety of novel antigens to the immune system, some of which could be exploited for immunotherapy.
Halder
T
T
Medical and Natural Sciences Research Center, Medical Hospital, University of Tübingen, Germany.
Pawelec
G
G
Kirkin
A F
AF
Zeuthen
J
J
Meyer
H E
HE
Kun
L
L
Kalbacher
H
H
D000818:Animals; D017306:Annexin A2 / Q000276:immunology; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D016466:CHO Cells; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D006683:HLA-DQ Antigens / Q000276:immunology; D006684:HLA-DR Antigens / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008545:Melanoma / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology; D009363:Neoplasm Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D014407:Tumor Cells, Cultured; D058951:gp100 Melanoma Antigen
D017306:Annexin A2; D000951:Antigens, Neoplasm; D006683:HLA-DQ Antigens; D006684:HLA-DR Antigens; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C548637:PMEL protein, human; D010446:Peptide Fragments; D058951:gp100 Melanoma Antigen