The effects of anti-asthma drugs on mediator release from cultured human mast cells

Background A method for generating human mast cells in vitro was recently e stablished. Little is known about the pharmacological profiles of allergic mediator release from cultured mast cells. Objective The main objective was to investigate the nature of cultured mast cells from a pharmacological point of view. We examined the effect of anti -asthma drugs on the release of histamine, sulfidoleukotrienes (LTs) and pr ostaglandin D-2 (PGD(2)) from the cultured mast cells. Methods Using the method established by Saito et al. we cultured cord blood mononuclear cells in the presence of 80 ng/mL stem cell factor (SCF), 50 n g/mL interleukin-6 (IL-6) and 300 nmol/L prostaglandin E-2 (PGE(2)), and ob tained almost pure (>99%) mast cells. We sensitized cultured mast cells wit h immunoglobulin E (IgE)-rich serum, and then treated them with some anti-a sthma drugs before challenge with anti-human IgE. Released histamine, LTs a nd PGD were measured by high-performance liquid chromatography, commercial enzyme-linked immunosorbent assay (ELISA) and enzyme immunoassay (EIA) syst ems, respectively. Results The cultured mast cells released histamine, LTs and PGD(2) followin g immunological stimulation through IgE. The mast cell stabilizing agents d isodium cromoglycate (DSCG, 1 mmol/L) and azelastine (100 mu mol/L) signifi cantly inhibited the release of these three mediators. The beta-adrenocepto r agonists isoproterenol, salbutamol, and clenbuterol also inhibited all th ree mediators' release in a concentration-dependent manner. The nonselectiv e and selective phosphodiesterase (PDE) inhibitors theophylline, rolipram, and cilostazol had no significant effect on mediator release at clinically useful concentrations. BAY x 1005 (a 5-lipoxygenase-activating protein inhi bitor) inhibited the LTs release, whereas indomethacin (a cyclo-oxygenase I and II inhibitor) and NS-398 (a cyclooxygenase II inhibitor) inhibited PGD (2) release. Conclusions The present results indicate that cultured mast cells release h istamine, LTs and PGD(2) following IgE crosslinking. Anti-asthma drugs show ed a characteristic suppression of the release of each mediator. The suppre ssive actions of these drugs are similar to their pharmacological actions o n human lung mast cells. These results suggest that cultured mast cells are useful for the analysis of function and pharmacological profiles of lung m ast cells.