MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection
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 molecule...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1294565/full |
| _version_ | 1797366570149740544 |
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| author | Marten Meyer Marten Meyer Marten Meyer Christina Parpoulas Titouan Barthélémy Jonas P. Becker Jonas P. Becker Pornpimol Charoentong Pornpimol Charoentong Pornpimol Charoentong Yanhong Lyu Selina Börsig Selina Börsig Nadja Bulbuc Claudia Tessmer Claudia Tessmer Lisa Weinacht David Ibberson Patrick Schmidt Patrick Schmidt Rüdiger Pipkorn Stefan B. Eichmüller Peter Steinberger Katharina Lindner Katharina Lindner Isabel Poschke Isabel Poschke Michael Platten Michael Platten Michael Platten Michael Platten Michael Platten Michael Platten Stefan Fröhling Stefan Fröhling Stefan Fröhling Angelika B. Riemer Angelika B. Riemer Jessica C. Hassel Maria Paula Roberti Maria Paula Roberti Dirk Jäger Dirk Jäger Inka Zörnig Inka Zörnig Frank Momburg Frank Momburg |
| author_facet | Marten Meyer Marten Meyer Marten Meyer Christina Parpoulas Titouan Barthélémy Jonas P. Becker Jonas P. Becker Pornpimol Charoentong Pornpimol Charoentong Pornpimol Charoentong Yanhong Lyu Selina Börsig Selina Börsig Nadja Bulbuc Claudia Tessmer Claudia Tessmer Lisa Weinacht David Ibberson Patrick Schmidt Patrick Schmidt Rüdiger Pipkorn Stefan B. Eichmüller Peter Steinberger Katharina Lindner Katharina Lindner Isabel Poschke Isabel Poschke Michael Platten Michael Platten Michael Platten Michael Platten Michael Platten Michael Platten Stefan Fröhling Stefan Fröhling Stefan Fröhling Angelika B. Riemer Angelika B. Riemer Jessica C. Hassel Maria Paula Roberti Maria Paula Roberti Dirk Jäger Dirk Jäger Inka Zörnig Inka Zörnig Frank Momburg Frank Momburg |
| author_sort | Marten Meyer |
| collection | DOAJ |
| description | 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 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. |
| first_indexed | 2024-03-08T17:05:39Z |
| format | Article |
| id | doaj.art-2562f0907ae7489481331f6d349419cb |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-08T17:05:39Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-2562f0907ae7489481331f6d349419cb2024-01-04T04:57:02ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-01-011410.3389/fimmu.2023.12945651294565MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detectionMarten Meyer0Marten Meyer1Marten Meyer2Christina Parpoulas3Titouan Barthélémy4Jonas P. Becker5Jonas P. Becker6Pornpimol Charoentong7Pornpimol Charoentong8Pornpimol Charoentong9Yanhong Lyu10Selina Börsig11Selina Börsig12Nadja Bulbuc13Claudia Tessmer14Claudia Tessmer15Lisa Weinacht16David Ibberson17Patrick Schmidt18Patrick Schmidt19Rüdiger Pipkorn20Stefan B. Eichmüller21Peter Steinberger22Katharina Lindner23Katharina Lindner24Isabel Poschke25Isabel Poschke26Michael Platten27Michael Platten28Michael Platten29Michael Platten30Michael Platten31Michael Platten32Stefan Fröhling33Stefan Fröhling34Stefan Fröhling35Angelika B. Riemer36Angelika B. Riemer37Jessica C. Hassel38Maria Paula Roberti39Maria Paula Roberti40Dirk Jäger41Dirk Jäger42Inka Zörnig43Inka Zörnig44Frank Momburg45Frank Momburg46Antigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyDivision of Immunotherapy and Immunoprevention, DKFZ, Heidelberg, GermanyGerman Center for Infection Research (DZIF) Partner Site Heidelberg, Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyCenter for Quantitative Analysis of Molecular and Cellular Biosystems (Bioquant), Heidelberg University, Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyDeep Sequencing Core Facility, Heidelberg University, Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyGMP and T Cell Therapy, DKFZ, Heidelberg, GermanyGMP and T Cell Therapy, DKFZ, Heidelberg, GermanyGMP and T Cell Therapy, DKFZ, Heidelberg, GermanyDivision of Immune Receptors and T Cell Activation, Center for Pathophysiology, Infectiology, Medical University of Vienna, Vienna, Austria0Clinical Cooperation Unit Neuroimmunology and Brain Tumor Immunology, DKFZ, Heidelberg, Germany1Immune Monitoring Unit, NCT Heidelberg and DKFZ, Heidelberg, Germany0Clinical Cooperation Unit Neuroimmunology and Brain Tumor Immunology, DKFZ, Heidelberg, Germany1Immune Monitoring Unit, NCT Heidelberg and DKFZ, Heidelberg, Germany0Clinical Cooperation Unit Neuroimmunology and Brain Tumor Immunology, DKFZ, Heidelberg, Germany1Immune Monitoring Unit, NCT Heidelberg and DKFZ, Heidelberg, Germany2German Cancer Consortium (DKTK), DKFZ, Core Center, Heidelberg, Germany3Department of Neurology, Medical Faculty Mannheim, Mannheim Center for Translational Neuroscience (MCTN), Heidelberg University, Mannheim, Germany4DKFZ Hector Cancer Institute at the University Medical Center, Mannheim, Germany5Helmholtz Institute for Translational Oncology, Mainz (HI-TRON Mainz), Mainz, Germany2German Cancer Consortium (DKTK), DKFZ, Core Center, Heidelberg, Germany6Division of Translational Medical Oncology, NCT Heidelberg and DKFZ, Heidelberg, Germany7Institute of Human Genetics, Heidelberg University, Heidelberg, GermanyDivision of Immunotherapy and Immunoprevention, DKFZ, Heidelberg, GermanyGerman Center for Infection Research (DZIF) Partner Site Heidelberg, Heidelberg, Germany8Section of DermatoOncology, Department of Dermatology and NCT, Heidelberg University Hospital, Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyClinical Cooperation Unit Applied Tumor Immunity, DKFZ, Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyAntigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, GermanyDepartment of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Heidelberg, GermanyPeptide-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.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1294565/fullT cellstumor immunotherapypeptide-MHC class I multimerneoepitope screeningT cell receptor discoverytumor neoantigen |
| spellingShingle | Marten Meyer Marten Meyer Marten Meyer Christina Parpoulas Titouan Barthélémy Jonas P. Becker Jonas P. Becker Pornpimol Charoentong Pornpimol Charoentong Pornpimol Charoentong Yanhong Lyu Selina Börsig Selina Börsig Nadja Bulbuc Claudia Tessmer Claudia Tessmer Lisa Weinacht David Ibberson Patrick Schmidt Patrick Schmidt Rüdiger Pipkorn Stefan B. Eichmüller Peter Steinberger Katharina Lindner Katharina Lindner Isabel Poschke Isabel Poschke Michael Platten Michael Platten Michael Platten Michael Platten Michael Platten Michael Platten Stefan Fröhling Stefan Fröhling Stefan Fröhling Angelika B. Riemer Angelika B. Riemer Jessica C. Hassel Maria Paula Roberti Maria Paula Roberti Dirk Jäger Dirk Jäger Inka Zörnig Inka Zörnig Frank Momburg Frank Momburg MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection Frontiers in Immunology T cells tumor immunotherapy peptide-MHC class I multimer neoepitope screening T cell receptor discovery tumor neoantigen |
| title | MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection |
| title_full | MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection |
| title_fullStr | MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection |
| title_full_unstemmed | MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection |
| title_short | MediMer: a versatile do-it-yourself peptide-receptive MHC class I multimer platform for tumor neoantigen-specific T cell detection |
| title_sort | medimer a versatile do it yourself peptide receptive mhc class i multimer platform for tumor neoantigen specific t cell detection |
| topic | T cells tumor immunotherapy peptide-MHC class I multimer neoepitope screening T cell receptor discovery tumor neoantigen |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1294565/full |
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