囊性纤维化患者的诱导痰：炎症标志物的周内再现性

Induced sputum in cystic fibrosis: within-week reproducibility of inflammatory markers.

作者信息

Smountas Argyrios A, Lands Larry C, Mohammed Shawn R, Grey Vijaylaxmi

机构信息

Department of Respiratory Medicine, McGill University Medical Centre, Montreal Children's Hospital, Montreal, 2300 Tupper Street, Montreal, Quebec, Canada H3H 1P3.

出版信息

Clin Biochem. 2004 Nov;37(11):1031-6. doi: 10.1016/j.clinbiochem.2004.07.008.

DOI:10.1016/j.clinbiochem.2004.07.008

PMID:15498533

Abstract

UNLABELLED

Analysis of induced sputum has provided significant insight into the inflammatory response in chronic respiratory diseases such as asthma. The thick, tenacious nature of cystic fibrosis (CF) sputum presents certain challenges to such evaluation. We describe the development of a methodology to assess CF sputum, and the within-week reproducibility (to limit the possibility of any change in clinical status) of cellular and inflammatory markers.

METHODS

Seventeen young adults [9 males, 8 females, mean age 24 (5), median (quartile range) years, percentage of predicted FEV(1) = 64.0 (18.0%)] with CF underwent sputum inductions on the Monday and Thursday of the same week. Patients were pretreated with 400 microg salbutamol and subsequently inhaled 5% saline via a breath enhanced nebulizer. Every 3-min nebulization was interrupted to allow for expectoration of sputum into a polypropylene pot. Sputum samples were dispersed with a solution of dithiothreitol (DTT) and deoxyribonuclease to allow for analysis of total cell count (TCC) and percentage neutrophils (%Neut). Measurement of tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), and neutrophil elastase was performed on samples dispersed with DTT alone.

RESULTS

There were no significant differences between the measurements taken in the same week. Values for Day 1 versus Day 2 were as follows: TCC 20.8 (6.4) vs. 17.6 (2.5) x 10(6) cells/ml; %Neut: 94.1 (0.0) vs. 95.4 (0.5) %; TNF-alpha 7.5 (26.0) vs. 21.0 (44.0) pg/ml; IL-8 610.0 (422.0) vs. 524.0 (587.0) ng/ml and neutrophil elastase 110.0 (19.75) vs. 49.75 (60.75) microM. High intraclass correlation coefficients (ICC) for TCC, %Neut, TNF-alpha IL-8 and neutrophil elastase were found (ICC = 0.76, 0.82, 0.93, 0.82, 0.74, respectively).

CONCLUSIONS

The method developed here for the analysis of CF sputum shows good reproducibility and can be used to evaluate therapeutic interventions in patients with cystic fibrosis.

摘要

未标注

诱导痰分析为深入了解哮喘等慢性呼吸道疾病的炎症反应提供了重要见解。囊性纤维化（CF）痰液质地浓稠、黏性大，给此类评估带来了一定挑战。我们描述了一种评估CF痰液的方法的开发过程，以及细胞和炎症标志物在一周内的可重复性（以限制临床状态发生任何变化的可能性）。

方法

17名患有CF的年轻成年人[9名男性，8名女性，平均年龄24（5）岁，中位数（四分位间距），预测FEV（1）百分比 = 64.0（18.0%）]在同一周的周一和周四进行了痰液诱导。患者先用400微克沙丁胺醇预处理，随后通过呼吸增强雾化器吸入5%盐水。每隔3分钟中断雾化，以便将痰液咳出到聚丙烯罐中。痰液样本用二硫苏糖醇（DTT）和脱氧核糖核酸酶溶液分散，以分析总细胞计数（TCC）和中性粒细胞百分比（%Neut）。仅用DTT分散的样本进行肿瘤坏死因子-α（TNF-α）、白细胞介素-8（IL-8）和中性粒细胞弹性蛋白酶的测量。

结果

同一周内的测量结果无显著差异。第1天与第2天的值如下：TCC 20.8（6.4）对17.6（2.5）×10（6）个细胞/毫升；%Neut：94.1（0.0）对95.4（0.5）%；TNF-α 7.5（26.0）对21.0（44.0）皮克/毫升；IL-8 610.0（422.0）对524.0（587.0）纳克/毫升；中性粒细胞弹性蛋白酶110.0（19.75）对49.75（60.75）微摩尔。TCC、%Neut、TNF-α、IL-8和中性粒细胞弹性蛋白酶的组内相关系数（ICC）较高（分别为ICC = 0.76、0.82、0.93、0.82、0.74）。