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000287071 1001_ $$0P:(DE-He78)27b8b96bde8e36bbbc5e91485b08411a$$aMeyer, Marten$$b0$$eFirst author$$udkfz
000287071 245__ $$aMediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection.
000287071 260__ $$aLausanne$$bFrontiers Media$$c2023
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000287071 500__ $$a#EA:D121#EA:D120#LA:D120# / HI-TRON / Front Immunol. 2023; 14: 1294565.Published online 2024 Jan 4. doi: 10.3389/fimmu.2023.1294565 / in 2023 mitgezählt
000287071 520__ $$aPeptide-loaded MHC class I (pMHC-I) multimers have revolutionized our capabilities to monitor disease-associated T cell responses with high sensitivity and specificity. To improve the discovery of T cell receptors (TCR) targeting neoantigens of individual tumor patients with recombinant MHC molecules, we developed a peptide-loadable MHC class I platform termed MediMer. MediMers are based on soluble disulfide-stabilized β2-microglobulin/heavy chain ectodomain single-chain dimers (dsSCD) that can be easily produced in large quantities in eukaryotic cells and tailored to individual patients' HLA allotypes with only little hands-on time. Upon transient expression in CHO-S cells together with ER-targeted BirA biotin ligase, biotinylated dsSCD are purified from the cell supernatant and are ready to use. We show that CHO-produced dsSCD are free of endogenous peptide ligands. Empty dsSCD from more than 30 different HLA-A,B,C allotypes, that were produced and validated so far, can be loaded with synthetic peptides matching the known binding criteria of the respective allotypes, and stored at low temperature without loss of binding activity. We demonstrate the usability of peptide-loaded dsSCD multimers for the detection of human antigen-specific T cells with comparable sensitivities as multimers generated with peptide-tethered β2m-HLA heavy chain single-chain trimers (SCT) and wild-type peptide-MHC-I complexes prior formed in small-scale refolding reactions. Using allotype-specific, fluorophore-labeled competitor peptides, we present a novel dsSCD-based peptide binding assay capable of interrogating large libraries of in silico predicted neoepitope peptides by flow cytometry in a high-throughput and rapid format. We discovered rare T cell populations with specificity for tumor neoepitopes and epitopes from shared tumor-associated antigens in peripheral blood of a melanoma patient including a so far unreported HLA-C*08:02-restricted NY-ESO-1-specific CD8+ T cell population. Two representative TCR of this T cell population, which could be of potential value for a broader spectrum of patients, were identified by dsSCD-guided single-cell sequencing and were validated by cognate pMHC-I multimer staining and functional responses to autologous peptide-pulsed antigen presenting cells. By deploying the technically accessible dsSCD MHC-I MediMer platform, we hope to significantly improve success rates for the discovery of personalized neoepitope-specific TCR in the future by being able to also cover rare HLA allotypes.
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000287071 650_7 $$2Other$$aT cell receptor discovery
000287071 650_7 $$2Other$$aT cells
000287071 650_7 $$2Other$$aneoepitope screening
000287071 650_7 $$2Other$$apeptide-MHC class I multimer
000287071 650_7 $$2Other$$apersonalized medicine
000287071 650_7 $$2Other$$atumor immunotherapy
000287071 650_7 $$2Other$$atumor neoantigen
000287071 7001_ $$0P:(DE-He78)0fb041569f2b7b4b3d9243cdcfe0c731$$aParpoulas, Christina$$b1$$eFirst author$$udkfz
000287071 7001_ $$0P:(DE-He78)62aef57e345d66b980567aaea5c45dde$$aBarthélémy, Titouan$$b2$$eFirst author
000287071 7001_ $$0P:(DE-He78)45aa4fa7b16296bd3454cc3b68c33fc1$$aBecker, Jonas P$$b3$$udkfz
000287071 7001_ $$0P:(DE-He78)8dde8d3920dc06c59314abdb4ac7d2e8$$aCharoentong, Pornpimol$$b4$$udkfz
000287071 7001_ $$0P:(DE-He78)558f7ddc9b9893d0f57eb74e243cc4fd$$aLyu, Yanhong$$b5$$udkfz
000287071 7001_ $$0P:(DE-He78)1da983ccc6dc19ba6486e74c3f1c2e53$$aBörsig, Selina$$b6$$udkfz
000287071 7001_ $$0P:(DE-He78)8a8d6fb28cd77106b895bffdaa346dcd$$aBulbuc, Nadja$$b7$$udkfz
000287071 7001_ $$0P:(DE-He78)44a33c775d0e27db79f8fd9e97a99e0a$$aTessmer, Claudia$$b8$$udkfz
000287071 7001_ $$0P:(DE-He78)76115030f757daacba6261a90aa8e536$$aWeinacht, Lisa$$b9$$udkfz
000287071 7001_ $$aIbberson, David$$b10
000287071 7001_ $$0P:(DE-He78)0a9ff2c74a3fef06f552be5bb95bfd2d$$aSchmidt, Patrick$$b11$$udkfz
000287071 7001_ $$0P:(DE-HGF)0$$aPipkorn, Rüdiger$$b12
000287071 7001_ $$0P:(DE-He78)23fb8cfffbf2aa8eee5d51af417ad944$$aEichmüller, Stefan B$$b13$$udkfz
000287071 7001_ $$aSteinberger, Peter$$b14
000287071 7001_ $$0P:(DE-He78)9c599f876c762bc78d289674c15ba4a5$$aLindner, Katharina$$b15$$udkfz
000287071 7001_ $$0P:(DE-He78)e9c55f46b4b06cf835834ee7e3e00db8$$aPoschke, Isabel$$b16$$udkfz
000287071 7001_ $$0P:(DE-He78)5ef8651b0f857b9c640aa5b1498c43b5$$aPlatten, Michael$$b17$$udkfz
000287071 7001_ $$0P:(DE-He78)f0144d171d26dbedb67c9db1df35629d$$aFröhling, Stefan$$b18$$udkfz
000287071 7001_ $$0P:(DE-He78)3743a1b712edca2ffa829b7096d7037e$$aRiemer, Angelika B$$b19$$udkfz
000287071 7001_ $$aHassel, Jessica C$$b20
000287071 7001_ $$0P:(DE-He78)72e6f8462f1b903eef99df71d9d8817b$$aRoberti, Maria Paula$$b21$$udkfz
000287071 7001_ $$0P:(DE-He78)ed0321409c9cde20b380ae663dbcefd1$$aJäger, Dirk$$b22$$udkfz
000287071 7001_ $$0P:(DE-He78)e00d6a13ce5ea2af7bde67bac1319dd3$$aZörnig, Inka$$b23$$udkfz
000287071 7001_ $$0P:(DE-He78)b2290261145f21c46f2d42783c69d104$$aMomburg, Frank$$b24$$eLast author$$udkfz
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