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Live Cell Labeling with Terpyridine Derivative Proligands to Measure Cytotoxicity Mediated by Immune Cells
Journal article   Open access   Peer reviewed

Live Cell Labeling with Terpyridine Derivative Proligands to Measure Cytotoxicity Mediated by Immune Cells

Yuki Sakai, Satoshi Mizuta, Asuka Kumagai, Mohammed S O Tagod, Hiroaki Senju, Tatsufumi Nakamura, Craig T Morita and Yoshimasa Tanaka
ChemMedChem, Vol.12(23), pp.2006-2013
12/07/2017
DOI: 10.1002/cmdc.201700626
PMCID: PMC5975248
PMID: 29110403
url
https://doi.org/10.1002/cmdc.201700626View
Published (Version of record) Open Access

Abstract

Immunotherapy using immune checkpoint inhibitors and CAR-T cells has revolutionized treatment for patients with malignant tumors. However, measuring tumor cell cytotoxicity mediated by immune effector cells in clinical laboratories has been difficult due to the requirement for radioactive substances. In this study, a series of novel terpyridine derivative proligands were synthesized, and a non-radioactive cellular cytotoxicity assay using the newly synthesized compounds was developed for use in preclinical and clinical studies for cancer immunotherapy. Once internalized into target cells, the compounds are hydrolyzed by esterases, resulting in the intracellular accumulation of the negatively charged terpyridine derivatives. When the labeled target cells are recognized and killed by immune effector cells, the integrity of the cell membrane is disrupted, and the terpyridine derivatives are released. Upon combining the culture supernatant with europium (Eu ), the cytotoxicity of immune effector cells for the target cells can be quantitatively determined by measuring the intensity of the Eu /ligand-derived time-resolved fluorescence. Thus, the assay developed in this study would facilitate the development of novel cancer immunotherapies.
Cell Survival - drug effects Pyridines - chemistry U937 Cells Humans K562 Cells Pyridines - chemical synthesis Killer Cells, Natural - immunology Ligands Molecular Structure Pyridines - pharmacology Cell Survival - immunology

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