Temperature-based MHC class-I multimer peptide exchange for human HLA-A, B and C
Abstract
T cell recognition of specific antigens presented by major histocompatibility complexes class-I (MHC-I) can play an important role during immune responses against pathogens and cancer cells. Detection of T cell immunity is based on assessing the presence of antigen-specific cytotoxic CD8+ T cells using MHC class-I (MHC-I) multimer technology. Previously we have designed conditional peptides for HLA-A*02:01, H-2Kb and HLA-E that form stable peptide-MHC-I-complexes at low temperatures and dissociate when exposed to a defined elevated temperature. The resulting conditional MHC-I complex can easily and without additional handling be exchanged with a peptide of interest, allowing to exchange peptides in a ready-to-use multimer and a high-throughput manner. Here we present data that this peptide-exchange technology is a general applicable, ready-to-use and fast approach to load many different peptides in MHC-I multimers for alleles of the HLA-A, HLA-B and HLA-C loci. We describe the development of conditional peptides for HLA-A*03:01, HLA-A*11:01, HLA-B*07:02 and HLA-C*07:02 that only form stable peptide-MHC-I complexes at low temperatures, allowing peptide exchange at higher defined temperature. We document the ease and flexibility of this technology by monitoring CD8+ T cell responses to virus-specific peptide-MHC complexes in patients.
Authors 15
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Leiden University Medical Center
Affiliation as printed
Department of Hematology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Dermatology, Leiden University Medical Center
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Anneloes van der Plas - van Duijn Aachen Department of Dermatology, Leiden University Medical Center
Leiden University Medical Center
Affiliation as printed
Department of Dermatology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Dermatology, Leiden University Medical Center
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Wesley Huisman Aachen
Leiden University · Leiden University Medical Center
Affiliation as printed
Leiden University Center for Infectious Diseases, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Immunology, Leiden University Medical Center
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Leiden University Medical Center · Oncode Institute
Affiliation as printed
Department of Cell and Chemical Biology, Oncode Institute, Leiden University Medical Center
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Leiden University Medical Center · Oncode Institute · Meander Medisch Centrum
Affiliation as printed
Department of Cell and Chemical Biology, Oncode Institute, Leiden University Medical Center
Laboratory for Clinical Chemistry and Hematology, Meander Medical Centre
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Leiden University Medical Center
Affiliation as printed
Department of Hematology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Hematology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Dermatology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark
Department of Hematology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark
Department of Dermatology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Hematology, Leiden University Medical Center
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Leiden University Medical Center
Affiliation as printed
Department of Dermatology, Leiden University Medical Center
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