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Leukotriene B4 Levels in Sputum From Asthma Patients

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Leukotriene B4 Levels in Sputum From Asthma Patients

Andrew Higham et al. ERJ Open Res.

Abstract

Poor asthma control is associated with increased airway neutrophils. Leukotriene B4 (LTB4) is a potent neutrophil chemoattractant. We examined the levels of LTB4 levels in the sputum of asthma patients and the relationship with disease severity. 47 asthma patients (categorised according to Global Initiative for Asthma treatment stage) and 12 healthy controls provided sputum samples that were processed first with PBS to obtain supernatants and secondly with dithiothreitol (DTT) to obtain supernatants. LTB4 levels were determined by ELISA. LTB4 levels were significantly higher in step 1 (steroid naïve) and step 3 (inhaled corticosteroid (ICS) plus long acting β-agonist) patients than step 2 patients (ICS alone) (p=0.02 and p=0.01, respectively). There was very good correlation when comparing PBS processed to DTT processed supernatants. High LTB4 levels were found in the sputum of asthmatics at step 3 despite ICS use.

Conflict of interest statement

can be found alongside this article at openres.ersjournals.com

Figures

FIGURE 1
FIGURE 1
A comparison between leukotriene B4 (LTB4) levels in the sputum of healthy controls and asthmatics. LTB4 was measured in a) PBS processed sputum supernatants from 12 healthy controls and 47 asthmatics (step 1: n=17, step 2: n=14, step 3: n=16) and b) dithiothreitol (DTT) processed sputum supernatants from 12 healthy controls and 40 asthmatics (step 1: n=16, step 2: n=12, step 3: n=12) by ELISA. *: p<0.05, significant difference in LTB4 levels.
FIGURE 2
FIGURE 2
Univariate Spearman's analysis between leukotriene B4 (LTB4) levels measured in PBS and dithiothreitol (DTT) processed sputum. Analysis was performed in sputum supernatants isolated from a) all study subjects (12 healthy controls and 40 asthmatics: (step 1: n=16, step 2: n=12, step 3: n=12) or from b) step 1 only.
FIGURE 3
FIGURE 3
Leukotriene B4 (LTB4) levels in asthmatic patients when stratified according to the sputum cellularity. LTB4 levels were measured in a) PBS processed sputum supernatants and b) dithiothreitol (DTT) processed sputum supernatants from patients who were neutrophilic (neut) (PBS: n=15, DTT: n=11), eosinophilic (eos) (PBS: n=8, DTT: n=8), mixed (PBS: n=5, DTT: n=3) or paucigranulocytic (pauci) (PBS: n=13, DTT: n=12).

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