Blockade of superoxide generation prevents high‐affinity immunoglobulin E receptor‐mediated release of allergic mediators by rat mast cell line and human basophils

Abstract

These results indicate that both RBL-2H3 cells and human basophils generate superoxide anion upon FcepsilonRI cross-linking either by antibody or by allergen challenge and that blockade of the generation prevents the release of allergic mediators. The findings strongly support the role of superoxide generation in the activation of mast cells and basophils under both physiological and pathological conditions. The findings suggest that drugs regulating the superoxide generation have potential therapeutic use for allergic disorders.

A Comparison of Reactive Oxygen Species Generation by Rat Peritoneal Macrophages and Mast Cells Using the Highly Sensitive Real-Time Chemiluminescent Probe Pholasin: Inhibition of Antigen-Induced Mast Cell Degranulation by Macrophage-Derived Hydrogen Peroxide

“…More recently, the mast cell line rat basophilic leukemia 2H3 (RBL-2H3) has been shown to release low-level ROS when stimulated with Ag or calcium ionophore (A23187) using the detection techniques of luminol and MCLA (38,39). Although this release was inhibitable by both DPI and SOD, no steady baseline readings were shown before stimulation, although appropriate controls were shown on stimulation.…”

FcεRI Signaling of Mast Cells Activates Intracellular Production of Hydrogen Peroxide: Role in the Regulation of Calcium Signals

“…LTC 4 release was determined as described previously (25,26). Briefly, cells were activated as described above, and the LTC 4 content in supernatants was determined by an LTC 4 ELISA kit (Cayman Chemical, Ann Arbor, MI) according to the manufacturer's protocol.…”

“…The RBL-2H3 cells obtained from the National Institute of Health Sciences (Japan Collection of Research Biosources; cell number JCRB0023) were grown in DMEM (Sigma-Aldrich) supplemented with 10% FCS (Life Technologies, Tokyo, Japan) in a 5% CO 2 -containing atmosphere as described previously (25,26). The RBL-2H3 cells were harvested by incubating them in HBSS-supplemented bicarbonate (pH7.4) containing 1 mM EDTA/0.25% trypsin for 5 min at 37°C.…”

“…The production of ROS is sensitive to diphenyleneiodonium (DPI), a broad-spectrum inhibitor of flavoprotein-containing oxidoreductases (25). Similar DPI-inhibitable ROS production occurs in human leukocytes under both physiological (upon Ag stimulation) and pathological (upon allergen challenge) conditions (26). Furthermore, blockade of ROS production by DPI or antioxidants such as (Ϫ)-epigallocatechin gallate suppresses IgE-mediated histamine release and leukotriene C 4 (LTC 4 ) (27).…”

“…We previously reported that the RBL-2H3 mast cell line produces ROS upon Fc⑀RI cross-linking and releases them into the extracellular spaces (25,26). The production of ROS is sensitive to diphenyleneiodonium (DPI), a broad-spectrum inhibitor of flavoprotein-containing oxidoreductases (25).…”