Alizadeh Zahra, Mazinani Marzieh, Mortaz Esmaeil, Fazlollahi Mohammad Reza, Adcock Ian, Moein Mostafa
Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran.
Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran AND Department of Immunology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Iran J Allergy Asthma Immunol. 2018 Feb;17(1):1-8.
Different phenotypes of asthma from mild to severe are categorized based on diverse clinical features. A guideline for the recognition and treatment of asthma has been provided by Global Initiative for Asthma (GINA). To control symptoms and prevent asthma exacerbation in most patients combinational therapy with inhaled corticosteroids (ICS) and a long acting B2-adrenreceptor agonist (LABA) are recommended. Understanding asthma phenotypes would be helpful to improve asthma diagnosis and treatment. The aim of this study was to verify glucocorticoid receptor glcococorticoid receptor (GR) nuclear translocation in CD4 T cells treated with fluticasone furoate (FF), vilanterol (V) and FF/V combination in severe asthmatic patients compare to patients with moderate asthma and healthy controls using Immunocytochemistry (ICC). After taking blood and separating PBMCs from each subject, CD4 T cells were isolated from PBMCs using CD4+ T cell isolation kit. Isolated CD4 T cells were cultured in presence of FF, V and FF/V combination for 1 hour and after cytocentrifugation, cells were incubated with anti GR-antibody and subsequently stained with FITC bound secondary antibody and GR nuclear translocation was observed under microscope. The results showed significant increasing in GR nuclear translocation in treated CD4 T cells from patients with moderate asthma and controls compare to those severe asthmatic patients, along with treating cells with FF/V combination no significant GR nuclear translocation was observed compare to that of using mono treatment of cells with FF and V. Based on our findings, it can be concluded different mechanisms are responsible for severe asthma and moderate asthma.
根据不同的临床特征,哮喘从轻度到重度可分为不同的表型。全球哮喘防治创议(GINA)提供了哮喘识别和治疗指南。为控制大多数患者的症状并预防哮喘发作,推荐吸入性糖皮质激素(ICS)与长效β2肾上腺素受体激动剂(LABA)联合治疗。了解哮喘表型有助于改善哮喘的诊断和治疗。本研究的目的是,通过免疫细胞化学(ICC),比较重度哮喘患者与中度哮喘患者及健康对照,观察糠酸氟替卡松(FF)、维兰特罗(V)及FF/V联合治疗后CD4 T细胞中糖皮质激素受体(GR)的核转位情况。从每个受试者采集血液并分离外周血单核细胞(PBMC)后,使用CD4+ T细胞分离试剂盒从PBMC中分离出CD4 T细胞。将分离出的CD4 T细胞在FF、V及FF/V联合存在的情况下培养1小时,细胞离心涂片后,与抗GR抗体孵育,随后用异硫氰酸荧光素(FITC)标记的二抗染色,并在显微镜下观察GR核转位情况。结果显示,与重度哮喘患者相比,中度哮喘患者和对照组经治疗的CD4 T细胞中GR核转位显著增加,而且与FF和V单药治疗细胞相比,FF/V联合治疗细胞未观察到显著的GR核转位。根据我们的研究结果,可以得出结论,重度哮喘和中度哮喘的发病机制不同。