Development of a highly sensitive platform for protein-protein interaction detection and regulation of T cell function

被引:1
|
作者
Hayashi, Hideki [1 ,2 ]
Mak, Tak Wah [3 ,4 ,5 ]
Tanaka, Yoshimasa
Kubo, Yoshinao [6 ]
Izumida, Mai [6 ]
Urae, Ryuji [7 ]
Matsuyama, Toshifumi [8 ]
机构
[1] Nagasaki Univ, Sch Med, Nagasaki 8528523, Japan
[2] Nagasaki Univ, Ctr Med Innovat, Nagasaki 8528588, Japan
[3] Univ Hlth Network, Princess Margaret Canc Ctr, Toronto, ON M5G 2M9, Canada
[4] Univ Hong Kong, Li Ka Shing Fac Med, Sch Clin Med, Dept Pathol, Hong Kong 999077, Peoples R China
[5] Hong Kong Sci Pk, Ctr Oncol & Immunol, Hong Kong 999077, Peoples R China
[6] Nagasaki Univ, Inst Trop Med, Dept Clin Med, Nagasaki 8528523, Japan
[7] Souseikai Clin Res Ctr, Fukuoka 8120025, Japan
[8] Nagasaki Univ, Grad Sch Sci, Dept Forens Pathol & Sci, Nagasaki 8528523, Japan
关键词
interferon signaling; interferon-; gamma; protein interaction; IL-2; T cell activation; REVEALS; SINGLE; TRANSCRIPTION; REPORTER; COMPLEX;
D O I
10.1073/pnas.2318190121
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We developed a highly sensitive assay for detecting protein-protein interaction using chimeric receptors comprising two molecules of interest in the extracellular domain edly more sensitive than the NanoBiT system, currently considered one of the best detection systems for protein interaction. Employing chimeric receptors with extracellular domains from the IFN gamma or IL- 2 receptor and the intracellular domains of IFNAR1/2, the IFNARRS system effectively identifies low IFN gamma or IL- 2 levels. Cells stably expressing these chimeric receptors responded to IFN gamma secreted by activated T cells following various stimuli, including a specific peptide- antigen. The activation signals were further enhanced by the expression of relevant genes, such as costimulators, via IFN- stimulated strated the capability to detect other cytokines by using the corresponding extracellular domains from these target cytokine receptors.
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页数:12
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