Journal article
Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells
Department of Health Technology, Technical University of Denmark1
Experimental & Translational Immunology, Department of Health Technology, Technical University of Denmark2
T-Cells and Cancer, Experimental & Translational Immunology, Department of Health Technology, Technical University of Denmark3
Constructor University4
University of Copenhagen5
European Molecular Biology Laboratory6
Technical University of Munich7
The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer–based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the a1 and a2 helices close to the F pocket.
Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2Kb can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules.
Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.
Language: | English |
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Year: | 2019 |
Pages: | eaau9039-eaau9039 |
ISSN: | 24709468 , 15578445 and 00652776 |
Types: | Journal article |
DOI: | 10.1126/sciimmunol.aau9039 |
ORCIDs: | Saini, Sunil Kumar , 0000-0003-0049-711X , 0000-0002-7169-5765 , 0000-0003-4966-9752 , 0000-0002-9451-6037 , 0000-0002-2227-8615 , 0000-0001-5163-2663 , 0000-0003-2872-6279 , 0000-0002-5527-6149 , Hadrup, Sine Reker , Tamhane, Tripti and Ramskov, Sofie |