A versatile T cell-based assay to assess therapeutic antigen-specific PD-1-targeted approaches
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Maarten Versteven1,*, Johan M.J. Van den Bergh1,*, Katrijn Broos2,*, Fumihiro Fujiki3,*, Diana Campillo-Davo1, Hans De Reu1, Soyoko Morimoto4, Quentin Lecocq2, Marleen Keyaerts5,6, Zwi Berneman1,7,8, Haruo Sugiyama3, Viggo F.I. Van Tendeloo1, Karine Breckpot2 and Eva Lion1,8
1Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, Antwerp, Belgium
2Laboratory for Molecular and Cellular Therapy, Vrije Universiteit Brussel, Brussels, Belgium
3Department of Cancer Immunology, Osaka University Graduate School of Medicine, Osaka, Japan
4Department of Cancer Immunotherapy, Osaka University Graduate School of Medicine, Osaka, Japan
5In Vivo Cellular and Molecular Imaging Laboratory, Vrije Universiteit Brussel, Brussels, Belgium
6Nuclear Medicine Department, UZ Brussel, Brussels, Belgium
7Division of Hematology, University Hospital Antwerp, Antwerp, Belgium
8Center for Cell Therapy and Regenerative Medicine, University Hospital Antwerp, Antwerp, Belgium
*These authors contributed equally to this work
Maarten Versteven, email: [email protected]
Keywords: PD-1/PD-L1 immune checkpoint pathway; antigen-specific; bioassay; flow cytometry; immune checkpoint inhibition
Received: January 16, 2018 Accepted: May 24, 2018 Published: June 12, 2018
Blockade of programmed cell death protein 1 (PD-1) immune checkpoint receptor signaling is an established standard treatment for many types of cancer and indications are expanding. Successful clinical trials using monoclonal antibodies targeting PD-1 signaling have boosted preclinical research, encouraging development of novel therapeutics. Standardized assays to evaluate their bioactivity, however, remain restricted. The robust bioassays available all lack antigen-specificity. Here, we developed an antigen-specific, short-term and high-throughput T cell assay with versatile readout possibilities. A genetically modified T cell receptor (TCR)-deficient T cell line was stably transduced with PD-1. Transfection with messenger RNA encoding a TCR of interest and subsequent overnight stimulation with antigen-presenting cells, results in eGFP-positive and granzyme B-producing T cells for single cell or bulk analysis. Control antigen-presenting cells induced reproducible high antigen-specific eGFP and granzyme B expression. Upon PD-1 interaction, ligand-positive antigen-presenting immune or tumor cells elicited significantly lower eGFP and granzyme B expression, which could be restored by anti-PD-(L)1 blocking antibodies. This convenient cell-based assay shows a valuable tool for translational and clinical research on antigen-specific checkpoint-targeted therapy approaches.
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