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

• 成果类型: Article
• 署名作者: Hayashi, Hideki; Mak, Tak Wah; Tanaka, Yoshimasa; Kubo, Yoshinao; Izumida, Mai; Urae, Ryuji; Matsuyama, Toshifumi
• 署名单位: Nagasaki University; Nagasaki University; University of Toronto; University Health Network Toronto; Princess Margaret Cancer Centre; University of Hong Kong; Nagasaki University; Nagasaki University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-13677
• DOI: 10.1073/pnas.2318190121
• 发表日期: 2024-08-13
• 关键词: reveals; single; transcription; reporter; complex

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.