TY - JOUR
T1 - Effect of sputum processing with dithiothreitol on the detection of inflammatory mediators in chronic bronchitis and bronchiectasis
AU - Woolhouse, Ian
AU - Bayley, Darren
AU - Stockley, Robert
PY - 2002/8/1
Y1 - 2002/8/1
N2 - BACKGROUND: Sputum analysis is used increasingly to assess airway inflammation in patients with chronic obstructive pulmonary disease, including those with chronic bronchitis and bronchiectasis. However, it is not known whether dithiothreitol (DTT), a reducing mucolytic agent regularly used to homogenise sputum, affects the detection of inflammatory mediators in the sputum soluble phase from such patients. METHODS: Thirty two spontaneous sputum samples were collected from 13 patients with chronic bronchitis and 17 with bronchiectasis. An aliquot from each sample was treated with either freshly prepared 0.1% DTT plus normal saline (NaCl) or NaCl alone, then ultracentrifuged to obtain the sputum sol phase. Interleukin (IL)-1beta, IL-6, IL-8, leukotriene B(4) (LTB(4)), secretory leukoprotease inhibitor (SLPI), alpha-1-antitrypsin (alpha(1)-AT), and tumour necrosis factor alpha (TNFalpha) were measured by ELISA, and neutrophil elastase (NE) and myeloperoxidase (MPO) by chromogenic substrate assay. The effect of DTT on the detection of assay standards was also determined. RESULTS: Median levels of IL-1beta, IL-6, IL-8, SLPI, and NE were similar in the DTT and NaCl treated samples. There was a significant reduction in median (IQR) levels of detectable TNFalpha (0.07 (0.00-0.47) pM v 0.90 (0.06-6.98) pM, p
AB - BACKGROUND: Sputum analysis is used increasingly to assess airway inflammation in patients with chronic obstructive pulmonary disease, including those with chronic bronchitis and bronchiectasis. However, it is not known whether dithiothreitol (DTT), a reducing mucolytic agent regularly used to homogenise sputum, affects the detection of inflammatory mediators in the sputum soluble phase from such patients. METHODS: Thirty two spontaneous sputum samples were collected from 13 patients with chronic bronchitis and 17 with bronchiectasis. An aliquot from each sample was treated with either freshly prepared 0.1% DTT plus normal saline (NaCl) or NaCl alone, then ultracentrifuged to obtain the sputum sol phase. Interleukin (IL)-1beta, IL-6, IL-8, leukotriene B(4) (LTB(4)), secretory leukoprotease inhibitor (SLPI), alpha-1-antitrypsin (alpha(1)-AT), and tumour necrosis factor alpha (TNFalpha) were measured by ELISA, and neutrophil elastase (NE) and myeloperoxidase (MPO) by chromogenic substrate assay. The effect of DTT on the detection of assay standards was also determined. RESULTS: Median levels of IL-1beta, IL-6, IL-8, SLPI, and NE were similar in the DTT and NaCl treated samples. There was a significant reduction in median (IQR) levels of detectable TNFalpha (0.07 (0.00-0.47) pM v 0.90 (0.06-6.98) pM, p
UR - http://www.scopus.com/inward/record.url?scp=0036341137&partnerID=8YFLogxK
U2 - 10.1136/thorax.57.8.667
DO - 10.1136/thorax.57.8.667
M3 - Article
C2 - 12149524
SN - 0040-6376
VL - 57
SP - 667
EP - 671
JO - Thorax
JF - Thorax
IS - 8
ER -