Interrogating ligand-receptor interactions using highly sensitive cellular biosensors

被引:4
作者
Funk M.A. [1 ]
Leitner J. [1 ]
Gerner M.C. [2 ]
Hammerler J.M. [2 ]
Salzer B. [3 ,4 ]
Lehner M. [3 ,4 ]
Battin C. [1 ]
Gumpelmair S. [1 ]
Stiasny K. [5 ]
Grabmeier-Pfistershammer K. [6 ]
Steinberger P. [1 ]
机构
[1] Center for Pathophysiology, Infectiology and Immunology, Institute of Immunology, Division for Immune Receptors and T cell activation, Medical University of Vienna, Vienna
[2] Division of Biomedical Science, University of Applied Sciences FH Campus Wien, Vienna
[3] St. Anna Children’s Cancer Research Institute, Vienna
[4] Christian Doppler Laboratory for Next Generation CAR T Cells, Vienna
[5] Center for Virology, Medical University of Vienna, Vienna
[6] Department of Dermatology, Medical University of Vienna, Vienna
基金
美国国家卫生研究院; 奥地利科学基金会;
关键词
D O I
10.1038/s41467-023-43589-1
中图分类号
学科分类号
摘要
Interactions of membrane-resident proteins are important targets for therapeutic interventions but most methods to study them are either costly, laborious or fail to reflect the physiologic interaction of membrane resident proteins in trans. Here we describe highly sensitive cellular biosensors as a tool to study receptor-ligand pairs. They consist of fluorescent reporter cells that express chimeric receptors harboring ectodomains of cell surface molecules and intracellular signaling domains. We show that a broad range of molecules can be integrated into this platform and we demonstrate its applicability to highly relevant research areas, including the characterization of immune checkpoints and the probing of cells for the presence of receptors or ligands. The platform is suitable to evaluate the interactions of viral proteins with host receptors and to test for neutralization capability of drugs or biological samples. Our results indicate that cellular biosensors have broad utility as a tool to study protein-interactions. © 2023, The Author(s).
引用
收藏
相关论文
共 81 条
[11]  
Vazquez-Lombardi R., Et al., High-throughput T cell receptor engineering by functional screening identifies candidates with enhanced potency and specificity ll High-throughput T cell receptor engineering by functional screening identifies candidates with enhanced potency and specificity, Immunity, 55, pp. 1953-1965.e11, (2022)
[12]  
Joglekar A.V., Et al., T cell antigen discovery via signaling and antigen-presenting bifunctional receptors, Nat. Methods 2019 162, 16, pp. 191-198, (2019)
[13]  
Irving B.A., Weiss A., Surface chimeric receptors as tools in study of lymphocyte activation, Methods Enzymol., 327, pp. 210-228, (2000)
[14]  
Eshhar Z., Waks T., Gross G., Schindler D.G., Specific activation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the gamma or zeta subunits of the immunoglobulin and T-cell receptors, Proc. Natl Acad. Sci., 90, pp. 720-724, (1993)
[15]  
June C.H., Sadelain M., Chimeric Antigen Receptor Therapy, N. Engl. J. Med., 379, pp. 64-73, (2018)
[16]  
Roberts M.R., Et al., Targeting of Human Immunodeficiency Virus-Infected Cells by CD8+ T Lymphocytes Armed With Universal T-Cell Receptors, Blood, 84, pp. 2878-2889, (1994)
[17]  
Ellebrecht C.T., Et al., Reengineering chimeric antigen receptor T cells for targeted therapy of autoimmune disease, Science, 353, pp. 179-184, (2016)
[18]  
Lee J., Et al., Antigen-specific B cell depletion for precision therapy of mucosal pemphigus vulgaris, J. Clin. Invest., 130, pp. 6317-6324, (2020)
[19]  
Jutz S., Et al., A cellular platform for the evaluation of immune checkpoint molecules, Oncotarget, 8, pp. 64892-64906, (2017)
[20]  
Jutz S., Et al., Assessment of costimulation and coinhibition in a triple parameter T cell reporter line: Simultaneous measurement of NF-κB, NFAT and AP-1, J. Immunol. Methods, 430, pp. 10-20, (2016)