High-throughput T-cell epitope discovery through MHC peptide exchange.
Recombinant major histocompatibility complex (MHC) class I molecules complexed with pathogen-specific or other disease-associated antigens have become essential reagents for the analysis of adaptive T-cell responses. However, conventional techniques for the production of recombinant peptide-MHC (pMHC) complexes are highly involved and thereby limit the use of pMHC complexes in terms of antigen diversity. To make pMHC-based techniques suitable for high-throughput analyses we developed an MHC peptide exchange technology based on the use of conditional MHC ligands. This technology enables the parallel production of thousands of different pMHC complexes within hours, allowing the development of high-throughput MHC-based assay systems to identify MHC ligands and cytotoxic T-cell responses. These high-throughput assays should prove valuable for the screening of entire disease-associated proteomes, including pathogen-encoded proteomes, tumor-associated antigens, and autoimmune antigens.
Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Sine Reker
SR
Hadrup
Mireille
M
Toebes
Boris
B
Rodenko
Arnold H
AH
Bakker
David A
DA
Egan
Huib
H
Ovaa
Ton N M
TN
Schumacher
D019921:Biotinylation; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D018604:Epitope Mapping / Q000191:economics / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005805:Genes, MHC Class I; D006801:Humans; D008024:Ligands; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology*; D011485:Protein Binding; D017510:Protein Folding; D014466:Ultraviolet Rays
D018984:Epitopes, T-Lymphocyte; D008024:Ligands; D019151:Peptide Library; D010455:Peptides