TY  - JOUR
AU  - Lindner, Katharina
AU  - Stange, Saskia
AU  - Hlawatsch, Julian
AU  - Hamberger, Moritz
AU  - Boschert, Tamara
AU  - Tan, Chin Leng
AU  - Hülsmeyer, Ingrid
AU  - Sachse, Samira Moreen
AU  - Jünger, Simone
AU  - Ernst, Alexander
AU  - Glatki, Olga
AU  - Vinogradova, Sofiya
AU  - Fleischhacker, Lena
AU  - Eichmüller, Stefan
AU  - Fröhling, Stefan
AU  - Hassel, Jessica C
AU  - Zörnig, Inka
AU  - Lyu, Yanhong
AU  - Huang, Zhiquin
AU  - Jäger, Dirk
AU  - Momburg, Frank
AU  - Meyer, Marten
AU  - Bunse, Lukas
AU  - Green, Edward
AU  - Platten, Michael
AU  - Poschke, Isabel
TI  - ESPEC-SUIT: a versatile and robust platform to identify and track antigen-specific T cell receptors in patients with cancer.
JO  - Journal for ImmunoTherapy of Cancer
VL  - 13
IS  - 9
SN  - 2051-1426
CY  - London
PB  - BioMed Central
M1  - DKFZ-2025-01944
SP  - e012216
PY  - 2025
N1  - #EA:D170#LA:D170#
AB  - 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
KW  - Humans
KW  - Neoplasms: immunology
KW  - Receptors, Antigen, T-Cell: immunology
KW  - Receptors, Antigen, T-Cell: metabolism
KW  - Receptors, Antigen, T-Cell: genetics
KW  - Antigens, Neoplasm: immunology
KW  - Female
KW  - T cell receptor - TCR (Other)
KW  - immunotherapy (Other)
KW  - intratumoral (Other)
KW  - solid tumor (Other)
KW  - vaccine (Other)
KW  - Receptors, Antigen, T-Cell (NLM Chemicals)
KW  - Antigens, Neoplasm (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:40983341
DO  - DOI:10.1136/jitc-2025-012216
UR  - https://inrepo02.dkfz.de/record/304838
ER  -