A rapid and sensitive assay based on particle analysis for cell degranulation detection in basophils and mast cells

Pharmacol Res. 2016 Sep:111:374-383. doi: 10.1016/j.phrs.2016.05.033. Epub 2016 Jun 21.

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.

Keywords: Basophils; Degranulation; Mast cells; Particles analysis; Sinomenine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basophil Degranulation Test / instrumentation
  • Basophil Degranulation Test / methods*
  • Basophils / drug effects
  • Basophils / immunology
  • Basophils / metabolism*
  • Biosensing Techniques*
  • Calcium / metabolism
  • Cell Degranulation* / drug effects
  • Cell Line, Tumor
  • Dynamic Light Scattering
  • Equipment Design
  • Exocytosis / drug effects
  • Female
  • Ionomycin / pharmacology
  • Mast Cells / drug effects
  • Mast Cells / immunology
  • Mast Cells / metabolism*
  • Morphinans / pharmacology
  • Predictive Value of Tests
  • Rats
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Time Factors
  • p-Methoxy-N-methylphenethylamine / pharmacology

Substances

  • Morphinans
  • p-Methoxy-N-methylphenethylamine
  • Ionomycin
  • sinomenine
  • Calcium