%0 Journal Article
%A Lindner, Katharina
%A Stange, Saskia
%A Hlawatsch, Julian
%A Hamberger, Moritz
%A Boschert, Tamara
%A Tan, Chin Leng
%A Hülsmeyer, Ingrid
%A Sachse, Samira Moreen
%A Jünger, Simone
%A Ernst, Alexander
%A Glatki, Olga
%A Vinogradova, Sofiya
%A Fleischhacker, Lena
%A Eichmüller, Stefan
%A Fröhling, Stefan
%A Hassel, Jessica C
%A Zörnig, Inka
%A Lyu, Yanhong
%A Huang, Zhiquin
%A Jäger, Dirk
%A Momburg, Frank
%A Meyer, Marten
%A Bunse, Lukas
%A Green, Edward
%A Platten, Michael
%A Poschke, Isabel
%T ESPEC-SUIT: a versatile and robust platform to identify and track antigen-specific T cell receptors in patients with cancer.
%J Journal for ImmunoTherapy of Cancer
%V 13
%N 9
%@ 2051-1426
%C London
%I BioMed Central
%M DKFZ-2025-01944
%P e012216
%D 2025
%Z #EA:D170#LA:D170#
%X Methods to identify and characterize antigen-reactive T cell receptors (TCRs) represent important tools to understand and exploit T cell responses in patients with cancer and beyond. Current methods are hampered by the rarity of individual T cell clones and limited applicability to monitor both major histocompatibility complex (MHC) class I-restricted and MHC class II-restricted responses, hence insufficiently reflecting the entire antigen-reactive repertoire of a patient. To obtain broad and deep insight into polyclonal, antigen-specific TCR repertoires, we developed the 'epitope-specific expansion culture with subsequent identification of TCRs' (ESPEC-SUIT) assay to identify and track antigen-specific TCRs.In vitro stimulation of peripheral blood mononuclear cells with (vaccine-targeted) neoantigens was verified in cytokine secretion assays and read-out by TCRβ repertoire sequencing (TCRseq). Candidate antigen-reactive clonotypes were defined by specific expansion in cultures stimulated with relevant antigen, followed by TCR cloning and validation in co-cultures of TCR transgenic effector cells and peptide-presenting targets. Using TCRseq information, candidate and validated clonotypes were traced and characterized in bulk and single-cell repertoire sequencing data of longitudinally collected blood samples and tumor tissue.In a cohort of 32 patients with cancer, we demonstrate that ESPEC-SUIT supports strong, robust and reproducible expansion of CD4+ and CD8+ T cells in response to various antigens. TCRseq revealed highly polyclonal neoepitope-specific T cell responses, which can be further characterized with respect to cross-reactivity, affinity or human leukocyte antigen (HLA) restriction. In a subcohort of 10 patients, we selected 341 ESPEC-SUIT-derived TCRs for cloning and in vitro functional validation from >2000 candidates and confirmed antigen-reactivity for >75
%K Humans
%K Neoplasms: immunology
%K Receptors, Antigen, T-Cell: immunology
%K Receptors, Antigen, T-Cell: metabolism
%K Receptors, Antigen, T-Cell: genetics
%K Antigens, Neoplasm: immunology
%K Female
%K T cell receptor - TCR (Other)
%K immunotherapy (Other)
%K intratumoral (Other)
%K solid tumor (Other)
%K vaccine (Other)
%K Receptors, Antigen, T-Cell (NLM Chemicals)
%K Antigens, Neoplasm (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:40983341
%R 10.1136/jitc-2025-012216
%U https://inrepo02.dkfz.de/record/304838