Journal article
Determination of human γδ T cell–mediated cytotoxicity using a non-radioactive assay system
Journal of immunological methods, Vol.466(C), pp.32-40
03/2019
DOI: 10.1016/j.jim.2019.01.003
PMCID: PMC6817948
PMID: 30654042
Abstract
The adoptive transfer of immune effector cells, such as CD8+ killer αβ T cells, γδ T cells, NK (natural killer) cells, and genetically–modified T cells, has been receiving increasing attention. It is essential to determine cellular cytotoxicity so as to monitor the function and quality of ex vivo–expanded immune effector cells before infusion. The most common method is the [51Cr]–sodium chromate release assay. It is, however, preferable to avoid the use of radioactive materials in clinical laboratories. In order to establish a non-radioactive alternative to the standard radioactive assay, we previously synthesized a chelate–forming prodrug (BM-HT) and demonstrated that a combination of BM-HT and europium (Eu3+) was useful to determine NK cell–mediated cytotoxicity. In the present study, we examined whether or not this improved assay system could be used to determine the cellular cytotoxicity exhibited by Vγ2Vδ2+ γδ T cells. In addition, we compared Eu3+ and terbium (Tb3+) in the measurement of cellular cytotoxicity. Our assay system using BM-HT could be used successfully for the analysis of both γδ T cell receptor (TCR)– and CD16–mediated cytotoxicity. When the intensity of fluorescence was compared between Eu3+ and Tb3+, Tb3+ chelate was more sensitive than Eu3+ chelate, suggesting that the detection system using Tb3+ is superior to Eu3+ when tumor cells are not efficiently labeled with BM-HT. The method established herein is expected to promote the development of novel adoptive cell therapies for cancer.
Details
- Title: Subtitle
- Determination of human γδ T cell–mediated cytotoxicity using a non-radioactive assay system
- Creators
- Mohammed S.O Tagod - Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, JapanSatoshi Mizuta - Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, JapanYuki Sakai - Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, JapanMasashi Iwasaki - Center for Innovation in Immunoregulative Technology and Therapeutics, Graduate School of Medicine, Kyoto University, Yoshidakonoe-Cho, Sakyo-Ku, Kyoto 606-8501, JapanKengo Shiraishi - Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, JapanHiroaki Senju - Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, JapanHiroshi Mukae - Department of Respiratory Medicine, Nagasaki University School of Medicine, 1-7-1 Sakamoto, Nagasaki 852-8501, JapanCraig T Morita - Department of Internal Medicine and the Interdisciplinary Graduate Program in Immunology, Iowa City Veterans Affairs Health Care System, University of Iowa Carver College of Medicine, Iowa City, IA 52246, USAYoshimasa Tanaka - Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan
- Resource Type
- Journal article
- Publication Details
- Journal of immunological methods, Vol.466(C), pp.32-40
- DOI
- 10.1016/j.jim.2019.01.003
- PMID
- 30654042
- PMCID
- PMC6817948
- NLM abbreviation
- J Immunol Methods
- ISSN
- 0022-1759
- eISSN
- 1872-7905
- Publisher
- Elsevier B.V
- Grant note
- DOI: 10.13039/100000054, name: National Cancer Institute, award: CA097274, P30CA086862; DOI: 10.13039/501100001700, name: Ministry of Education, Culture, Sports, Science and Technology, award: 16K08844; DOI: 10.13039/100000738, name: U.S. Department of Veterans Affairs; DOI: 10.13039/100006379, name: Office of Research and Development; DOI: 10.13039/100007217, name: Health Services Research and Development; DOI: 10.13039/100007496, name: Biomedical Laboratory Research and Development, VA Office of Research and Development, award: 1 I01 BX000972-01A1; DOI: 10.13039/100009619, name: Japan Agency for Medical Research and Development, award: A48
- Language
- English
- Date published
- 03/2019
- Academic Unit
- Immunology; Internal Medicine
- Record Identifier
- 9984094370602771
Metrics
17 Record Views