%0 Journal Article
%A Wellach, Kathrin
%A Riemer, Angelika
%T Highly sensitive live-cell imaging-based cytotoxicity assay enables functional validation of rare epitope-specific CTLs.
%J Frontiers in immunology
%V 16
%@ 1664-3224
%C Lausanne
%I Frontiers Media
%M DKFZ-2025-01088
%P 1558620
%D 2025
%Z #EA:D410#LA:D410#
%X Many immunotherapeutic approaches aim to induce epitope-specific T-cell cytotoxicity. However, the identification-and especially the functional validation-of suitable epitopes by in vitro cytotoxicity assays can be challenging, particularly when the number of available epitope-specific cytotoxic T cells (CTLs) is limited. Here, we present a highly sensitive image-based cytotoxicity assay that allows the functional analysis of rare epitope-specific T cells. The live-cell imaging-based setup combines transient red labeling of target cells with a green caspase 3/7 probe, allowing reliable measurement of the fraction of apoptotic target cells. Time-course analysis enables the monitoring of subtle differences. This highly flexible assay can be applied to assess the killing of either target cells with endogenous epitope presentation or those artificially loaded with the epitope of interest. Analysis of assay sensitivity demonstrated that cytotoxicity mediated by as few as 0.1
%K Humans
%K T-Lymphocytes, Cytotoxic: immunology
%K T-Lymphocytes, Cytotoxic: metabolism
%K Epitopes, T-Lymphocyte: immunology
%K Cytotoxicity, Immunologic
%K Lymphocyte Activation Gene 3 Protein
%K Cytotoxicity Tests, Immunologic: methods
%K Cell Line, Tumor
%K Interferon-gamma: metabolism
%K Programmed Cell Death 1 Receptor: metabolism
%K Programmed Cell Death 1 Receptor: immunology
%K Apoptosis
%K Caspase 3: metabolism
%K T cells (Other)
%K cytotoxicity (Other)
%K epitopes (Other)
%K live-cell imaging (Other)
%K rare CTL populations (Other)
%K Epitopes, T-Lymphocyte (NLM Chemicals)
%K Lymphocyte Activation Gene 3 Protein (NLM Chemicals)
%K Interferon-gamma (NLM Chemicals)
%K Lag3 protein, human (NLM Chemicals)
%K Programmed Cell Death 1 Receptor (NLM Chemicals)
%K Caspase 3 (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:40406125
%2 pmc:PMC12095279
%R 10.3389/fimmu.2025.1558620
%U https://inrepo02.dkfz.de/record/301580