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MHC multimer technology has been developed for the detection, isolation, and characterization of T cells in infection, autoimmunity, and cancer. Here, we present a simple, fast, flexible, and efficient method to generate many different MHC class I (MHC I) multimers in parallel using temperature-mediated peptide exchange. We designed conditional peptides for HLA-A*02:01 and H-2Kb that form stable peptide\u2013MHC I complexes at low temperatures, but dissociate when exposed to a defined elevated temperature. The resulting conditional MHC I complexes, either alone or prepared as ready-to-use multimers, can swiftly be loaded with peptides of choice without additional handling and within a short time frame. We demonstrate the ease and flexibility of this approach by monitoring the antiviral immune constitution in an allogeneic stem cell transplant recipient and by analyzing CD8+ T cell responses to viral epitopes in mice infected with lymphocytic choriomeningitis virus or cytomegalovirus.<\/jats:p>","DOI":"10.1084\/jem.20180156","type":"journal-article","created":{"date-parts":[[2018,4,17]],"date-time":"2018-04-17T12:45:10Z","timestamp":1523969110000},"page":"1493-1504","update-policy":"https:\/\/doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":54,"title":["A flexible MHC class I multimer loading system for large-scale detection of antigen-specific T cells"],"prefix":"10.1084","volume":"215","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9742-929X","authenticated-orcid":false,"given":"Jolien J.","family":"Luimstra","sequence":"first","affiliation":[{"name":"Oncode Institute and Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands 1"},{"name":"Department of Cell Biology II, Netherlands Cancer Institute, Amsterdam, Netherlands 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2896-7275","authenticated-orcid":false,"given":"Malgorzata A.","family":"Garstka","sequence":"additional","affiliation":[{"name":"Core Research Lab, the Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, China 2"},{"name":"Department of Cell Biology II, Netherlands Cancer Institute, Amsterdam, Netherlands 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5101-2154","authenticated-orcid":true,"given":"Marthe C.J.","family":"Roex","sequence":"additional","affiliation":[{"name":"Department of Hematology, Leiden University Medical Center, Leiden, Netherlands 6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anke","family":"Redeker","sequence":"additional","affiliation":[{"name":"Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, Netherlands 4"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George M.C.","family":"Janssen","sequence":"additional","affiliation":[{"name":"Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, Netherlands 5"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter A.","family":"van Veelen","sequence":"additional","affiliation":[{"name":"Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, Netherlands 5"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5058-4110","authenticated-orcid":true,"given":"Ramon","family":"Arens","sequence":"additional","affiliation":[{"name":"Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, Netherlands 4"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.H. 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