صورة
Image_2_MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection.jpeg
| العنوان: | Image_2_MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection.jpeg |
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| المؤلفون: | Marten Meyer, Christina Parpoulas, Titouan Barthélémy, Jonas P. Becker, Pornpimol Charoentong, Yanhong Lyu, Selina Börsig, Nadja Bulbuc, Claudia Tessmer, Lisa Weinacht, David Ibberson, Patrick Schmidt, Rüdiger Pipkorn, Stefan B. Eichmüller, Peter Steinberger, Katharina Lindner, Isabel Poschke, Michael Platten, Stefan Fröhling, Angelika B. Riemer, Jessica C. Hassel, Maria Paula Roberti, Dirk Jäger, Inka Zörnig, Frank Momburg |
| سنة النشر: | 2024 |
| مصطلحات موضوعية: | Immunology, Applied Immunology (incl. Antibody Engineering, Xenotransplantation and T-cell Therapies), Autoimmunity, Cellular Immunology, Humoural Immunology and Immunochemistry, Immunogenetics (incl. Genetic Immunology), Innate Immunity, Transplantation Immunology, Tumour Immunology, Immunology not elsewhere classified, Genetic Immunology, Animal Immunology, Veterinary Immunology, T cells, tumor immunotherapy, peptide-MHC class I multimer, neoepitope screening, T cell receptor discovery, tumor neoantigen, personalized medicine |
| الوصف: | Peptide-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 β 2 -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 β 2 m-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 ... |
| نوع الوثيقة: | still image |
| اللغة: | unknown |
| العلاقة: | https://figshare.com/articles/figure/Image_2_MediMer_a_versatile_do-it-yourself_peptide-receptive_MHC_class_I_multimer_platform_for_tumor_neoantigen-specific_T_cell_detection_jpeg/24940758Test |
| DOI: | 10.3389/fimmu.2023.1294565.s002 |
| الإتاحة: | https://doi.org/10.3389/fimmu.2023.1294565.s002Test https://figshare.com/articles/figure/Image_2_MediMer_a_versatile_do-it-yourself_peptide-receptive_MHC_class_I_multimer_platform_for_tumor_neoantigen-specific_T_cell_detection_jpeg/24940758Test |
| حقوق: | CC BY 4.0 |
| رقم الانضمام: | edsbas.D8E6180E |
| قاعدة البيانات: | BASE |
| DOI: | 10.3389/fimmu.2023.1294565.s002 |
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