Effect Of Dexamethasone And Cyclosporin On Human Basophils: A Flow-assisted In Vitro Assay

BackgroundAllergic disorders are common,of which the treatment remains a challenge. Basophils,as effector cells of allergy,become a promising target of drugs.In vitro mediator and/or cytokine release tests have been used in the study of basophil activation.However,these tests are time consuming with low sensitivity and specificity.Newly established flow-assisted basophil activation test becomes a better way to assess the function of basophils in vitro,this technique is convenient and fast with satisfactory sensitivity and specificity.ObjectivesTo establish an in vitro model of IgE-mediated basophil activation.In order to investigate the effect of dexamethasone,cyclosporin and chlorphenamine on basophils in vitro,we assessed activation marker on the surface of basophils stimulated by anti-IgE antibody.MethodsPeripheral venous blood was incubated with different concentrations of dexamethasone,cyclosporin and chlorphenamine for 1 hour at 37℃followed by stimulation with anti-IgE antibody.Negative controls and positive controls were incubated with PBS under the same condition.Cells activated with anti-IgE antibody were served as positive controls.After labeling with CD63 FITC/CD123 PE/HLA-DR PerCP monoclonal antibodies,the CD63 on the surface of basophils were assessed by flow cytometry.Results1.The CD63 positive basophils were 0.70±0.86%without stimulation,which increased to 50.58±9.04%when stimulated with anti-IgE antibody.The difference issignifigant(P＜0.01).2.CD63 positive basophils incubated by dexamethasone in varying concentrations decreased significantly compared with positive control(P＜0.01).The inhibiton effect is dose-dependent.3.CD63 positive basophils incubated by cyclosporin in different concentrations decreased significantly compared with positive control(P＜0.01 ).There is no signifigant difference among different concentrations(P＞0.05).4.The percentage of CD63 positive basophils incubated by chlorphenamine in the concentration of 10^-2M was smaller than positive control(P＜0.05).There is no inhibitory effect in the concentration of 10^-3M and 10^-4M(P＞0.05).ConclusionFlow-assisted basophil activation in vitro is a novel test to assess the effect of drugs on basophils,which can be applied in exploring mechanisms of drugs and evaluating the efficacy of allergy treatment.Anti-IgE antibody induced basophil activation can be inhibited by dexamethasone and cyclosporine.The inhibition of chlorphenamine on basophils is not as strong as dexamethasone.