Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1,-A3,-A11, and-B7

AH Bakker, R Hoppes, C Linnemann… - Proceedings of the …, 2008 - National Acad Sciences
AH Bakker, R Hoppes, C Linnemann, M Toebes, B Rodenko, CR Berkers, SR Hadrup…
Proceedings of the National Academy of Sciences, 2008National Acad Sciences
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 …
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.
National Acad Sciences