Inflammatory Cell Biology Group, Division of Infection, Inflammation, and Immunity, Sir Henry Wellcome Laboratories, South Block, Southampton General Hospital, Southampton, UK.
Am J Respir Crit Care Med. 2010 May 15;181(10):1049-60. doi: 10.1164/rccm.200906-0857OC. Epub 2010 Jan 28.
Much effort is being made to discover noninvasive biomarkers of chronic airway disease that might enable better management, predict prognosis, and provide new therapeutic targets.
To undertake a comprehensive, unbiased proteomic analysis of induced sputum and identify novel noninvasive biomarkers for chronic obstructive pulmonary disease (COPD).
Induced sputum was obtained from patients with COPD with a spectrum of disease severity and from control subjects. Two-dimensional gel electrophoresis and mass spectrometric identification of differentially expressed proteins were first applied to induced sputum from patients with GOLD stage 2 COPD and healthy smoker control subjects. Initial results thus obtained were validated by a combination of immunoassays (Western blotting and ELISA) applied to a large subject cohort. The biomarkers were localized to bronchial mucosa by immunohistochemistry.
Of 1,325 individual protein spots identified, 37 were quantitatively and 3 qualitatively different between the two groups (P < 0.05%). Forty protein spots were subjected to tandem mass spectrometry, which identified 15 separate protein species. Seven of these were further quantified in induced sputum from 97 individuals. Using this sequential approach, two of these potential biomarkers (apolipoprotein A1 and lipocalin-1) were found to be significantly reduced in patients with COPD when compared with healthy smokers. Their levels correlated with FEV(1)/FVC, indicating their relationship to disease severity.
A potential role for apolipoprotein A1 and lipocalin-1 in innate defense has been postulated previously; our discovery of their reduction in COPD indicates a deficient innate defense system in airway disease that could explain increased susceptibility to infectious exacerbations.
目前正在进行大量努力以发现慢性气道疾病的非侵入性生物标志物，这些标志物可能使疾病得到更好的管理，预测预后，并提供新的治疗靶点。
对诱导痰进行全面，无偏的蛋白质组学分析，并确定慢性阻塞性肺疾病（COPD）的新型非侵入性生物标志物。
从患有COPD的患者（具有不同严重程度的疾病）和对照者中获得诱导痰。首先将差异表达蛋白的二维凝胶电泳和质谱鉴定应用于 GOLD 分期 2 COPD 患者和健康吸烟者对照者的诱导痰中。通过Western 印迹和 ELISA 免疫测定组合应用于大型研究对象队列，对由此获得的初始结果进行验证。通过免疫组织化学将生物标志物定位到支气管黏膜。
在鉴定的 1325 个单个蛋白斑点中，有 37 个在两组之间在数量上和 3 个在质量上存在差异（P <0.05％）。将 40 个蛋白斑点进行串联质谱分析，鉴定出 15 种不同的蛋白种类。其中有 7 种在 97 例个体的诱导痰中进一步进行了定量。使用这种顺序方法，在 COPD 患者中发现其中两种潜在的生物标志物（载脂蛋白 A1 和脂联素-1）的水平明显低于健康吸烟者。它们的水平与 FEV1/FVC 相关，表明与疾病严重程度有关。
先前已经提出了载脂蛋白 A1 和脂联素-1在先天防御中的潜在作用；我们在 COPD 中发现其减少表明气道疾病中先天防御系统不足，这可能解释了对感染性加重的易感性增加。
