Temperature-based MHC class-I multimer peptide exchange for human HLA-A, B and C

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. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/630039v1_ufig1.gif" ALT="Figure 1"> View larger version (17K): org.highwire.dtl.DTLVardef@10e6564org.highwire.dtl.DTLVardef@229e51org.highwire.dtl.DTLVardef@c7f7b5org.highwire.dtl.DTLVardef@57b688_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIT cell immunity relies on antigen-specific CD8+ T cells recognizing peptide MHC-I complexes. C_LIO_LIEstablishing temperature-based peptide exchange across multiple HLA alleles, resulting in a robust, easy, and fast system to generate peptide MHC-I complexes. C_LIO_LITemperature-based MHC class-I multimer demonstrate applicability across major MHC-I gene families for monitoring CD8+ T cell responses. C_LIO_LIEasy high-throughput peptide exchange potential, enhancing clinical utility of MHC multimer technology. C_LI