title: A rapid and sensitive assay based on particle analysis for cell degranulation detection in basophils and mast cells.
abstract: The degranulation of mast cells and basophils is often initiated by a number of pathophysiological responses, especially in allergic and inflammatory conditions. Efficient techniques and methods for determining the level of such degranulation are highly demanded for laboratory and clinical studies. In this work, a rapid and sensitive assay based on the particle analysis of granules in RBL-2H3 cells, a cell line widely used as a convenient model system to study the degranulation of mast cells and basophils, was developed to detect cell degranulation using a Nanosight NS300 in light scatter mode and dynamic light scattering (DLS) on a Malvern Zetasizer Nano-ZS instrument. Using this method, drug-induced mast cell degranulation and systemic anaphylaxis were efficiently determined both in cell culture medium and blood samples from animals in the current study. This promising method is expected to be widely used for screening anti-allergic and anti-inflammatory drugs both in vitro and in vivo models, as well as for determining the level of mast cell degranulation of the patients in the clinic.
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China.
authors.forename: Lufen
authors.initials: L
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Pi
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China.
authors.forename: Jianlin
authors.initials: J
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China; Department of Chemistry, Jinan University, Guangzhou, China.
authors.forename: Jiye
authors.initials: J
authors.lastname: Cai
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China. Electronic address: tli@must.edu.mo.
authors.forename: Ting
authors.initials: T
authors.lastname: Li
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China. Electronic address: lliu@must.edu.mo.
authors.forename: Liang
authors.initials: L
authors.lastname: Liu
mesh_terms: D000818:Animals; D015549:Basophil Degranulation Test / Q000295:instrumentation / Q000379:methods*; D001491:Basophils / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism; D015550:Cell Degranulation / Q000187:drug effects; D045744:Cell Line, Tumor; D000067493:Dynamic Light Scattering; D004867:Equipment Design; D005089:Exocytosis / Q000187:drug effects; D005260:Female; D015759:Ionomycin / Q000494:pharmacology; D008407:Mast Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D009019:Morphinans / Q000494:pharmacology; D011237:Predictive Value of Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013997:Time Factors; D003189:p-Methoxy-N-methylphenethylamine / Q000494:pharmacology
chemical_list: D009019:Morphinans; D003189:p-Methoxy-N-methylphenethylamine; D015759:Ionomycin; C009271:sinomenine; D002118:Calcium
keywords: Basophils; Degranulation; Mast cells; Particles analysis; Sinomenine