Barck Charlotte, Lundahl Joachim, Halldén Gunilla, Bylin Gunnar
Division of Respiratory Medicine and Allergology, Karolinska Institutet, Department of Medicine, Karolinska University Hospital Huddinge, Stockholm, Sweden.
Ann Allergy Asthma Immunol. 2005 Jul;95(1):86-92. doi: 10.1016/S1081-1206(10)61193-3.
Counting eosinophils in induced sputum seems to be a better way to survey disease activity than indirect clinical assessments of airway inflammation, such as pulmonary function measurement. The conventional analysis of induced sputum, however, is time-consuming and requires skilled personnel, which have restricted its use.
To determine whether measuring eosinophil cationic protein (ECP) levels in the entire sputum sample could replace the method of counting eosinophils and measuring ECP levels in the supernatant only to establish a clinically more applicable method of studying the intraindividual changes in eosinophilic activity in induced sputum.
In 13 patients with mild, nonsymptomatic allergic asthma, sputum was induced before and 24 hours after allergen challenge. The entire sputum sample was diluted with 0.1% dithiothreitol, incubated, and divided into 2 parts. One part was processed according to the conventional method, and released ECP levels in the supernatant were measured. The second part was treated with a lysing reagent. Cell debris was separated, and total (intracellular and extracellular) ECP levels in the solution were measured.
We found good correlation between total ECP levels in the entire sputum sample and released ECP levels in the supernatant before (r = 0.97) and 24 hours after (r = 0.99) allergen challenge (P < .01 for both). We also found a good correlation between the changes in total and released ECP levels (r = 0.99; P < .01).
Total ECP concentration seems to reflect the eosinophilic inflammatory changes in asthma and might be a useful tool in clinical practice.
相较于气道炎症的间接临床评估方法,如肺功能测量,诱导痰中嗜酸性粒细胞计数似乎是一种更好的疾病活动监测方式。然而,传统的诱导痰分析方法耗时且需要专业人员,这限制了其应用。
确定测量整个痰样本中嗜酸性粒细胞阳离子蛋白(ECP)水平是否可替代仅对上清液中的嗜酸性粒细胞进行计数并测量ECP水平的方法,以建立一种临床上更适用的研究诱导痰中嗜酸性粒细胞活性个体内变化的方法。
对13例轻度无症状过敏性哮喘患者,在变应原激发前及激发后24小时诱导痰液。将整个痰样本用0.1%二硫苏糖醇稀释、孵育,然后分为两部分。一部分按照传统方法处理,测量上清液中释放的ECP水平。第二部分用裂解试剂处理。分离细胞碎片,测量溶液中总的(细胞内和细胞外)ECP水平。
我们发现整个痰样本中总的ECP水平与变应原激发前(r = 0.97)及激发后24小时(r = 0.99)上清液中释放的ECP水平之间具有良好的相关性(两者P均 <.01)。我们还发现总的和释放的ECP水平变化之间具有良好的相关性(r = 0.99;P <.01)。
总的ECP浓度似乎可反映哮喘中的嗜酸性粒细胞炎症变化,可能是临床实践中的一种有用工具。