title: Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7.
abstract: Major histocompatibility complex (MHC) class I multimer technology has become an indispensable immunological assay system to dissect antigen-specific cytotoxic CD8(+) T cell responses by flow cytometry. However, the development of high-throughput assay systems, in which T cell responses against a multitude of epitopes are analyzed, has been precluded by the fact that for each T cell epitope, a separate in vitro MHC refolding reaction is required. We have recently demonstrated that conditional ligands that disintegrate upon exposure to long-wavelength UV light can be designed for the human MHC molecule HLA-A2. To determine whether this peptide-exchange technology can be developed into a generally applicable approach for high throughput MHC based applications we set out to design conditional ligands for the human MHC gene products HLA-A1, -A3, -A11, and -B7. Here, we describe the development and characterization of conditional ligands for this set of human MHC molecules and apply the peptide-exchange technology to identify melanoma-associated peptides that bind to HLA-A3 with high affinity. The conditional ligand technology developed here will allow high-throughput MHC-based analysis of cytotoxic T cell immunity in the vast majority of Western European individuals.
authors.affiliation: Divisions of Immunology and Cellular Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.
authors.forename: Arnold H
authors.initials: AH
authors.lastname: Bakker
authors.forename: Rieuwert
authors.initials: R
authors.lastname: Hoppes
authors.forename: Carsten
authors.initials: C
authors.lastname: Linnemann
authors.forename: Mireille
authors.initials: M
authors.lastname: Toebes
authors.forename: Boris
authors.initials: B
authors.lastname: Rodenko
authors.forename: Celia R
authors.initials: CR
authors.lastname: Berkers
authors.forename: Sine Reker
authors.initials: SR
authors.lastname: Hadrup
authors.forename: Wim J E
authors.initials: WJ
authors.lastname: van Esch
authors.forename: Mirjam H M
authors.initials: MH
authors.lastname: Heemskerk
authors.forename: Huib
authors.initials: H
authors.lastname: Ovaa
authors.forename: Ton N M
authors.initials: TN
authors.lastname: Schumacher
mesh_terms: D000483:Alleles; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002999:Clone Cells; D000939:Epitopes / Q000276:immunology; D015234:HLA-A Antigens / Q000276:immunology*; D015788:HLA-A1 Antigen / Q000276:immunology; D059870:HLA-A11 Antigen; D015791:HLA-A3 Antigen / Q000276:immunology; D015793:HLA-B7 Antigen / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008024:Ligands; D008545:Melanoma / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D014466:Ultraviolet Rays
chemical_list: D000939:Epitopes; D015234:HLA-A Antigens; D015788:HLA-A1 Antigen; D059870:HLA-A11 Antigen; D015791:HLA-A3 Antigen; D015793:HLA-B7 Antigen; D015395:Histocompatibility Antigens Class I; D008024:Ligands; D010455:Peptides