Identification and quantification of antigens associated with HLA class I molecules on a bladder tumour cell line. Implications for vaccine design.
The aim of this study was to isolate and identify antigenic peptides associated with HLA class I molecules on a bladder tumour cell line. HLA-A1 molecules were purified using an immunobead-purification technique and following elution of nonapeptides associated with the complex, their HPLC profile was determined by Tandem mass spectrometry (MS/MS). Three peptides were identified namely: (1) P991 (VTDPGNLLY); (2) P1041.5 (LTDLGFLVY), and (3) P1057.7 (LTDPHLLSY); these sequences matched elements of hepatitis B, MAGE1-A1 and herpes simplex viruses. These antigens had half-lives of approximately 120 min which is within the theoretical range of such short peptides. These results indicate that identification of MHC-associated peptides is possible without using an assay for cytotoxic T cells. Although this approach was applied on a relatively small scale, broader applications can be foreseen.
Torabi-Pour
Nasir
N
Uro/Oncology Research and Department of Immunology, St. Bartholomew's and The Royal London School of Medicine and Dentistry, London EC1A 7BE, UK.
Morrow
W John W
WJ
Saffie
Roghieh
R
Gowda
Prashant G
PG
Perrett
D
D
Oliver
Timothy R D
TR
D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D015788:HLA-A1 Antigen / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D009842:Oligopeptides / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000276:immunology*
D015788:HLA-A1 Antigen; D009842:Oligopeptides; D010446:Peptide Fragments