P.S. Fitch

Exhaled nitric oxide correlates with airway eosinophils in childhood asthma

Background: Exhaled nitric oxide has been proposed as a marker for airway inflammation in asthma. The aim of this study was to compare exhaled nitric oxide levels with inflammatory cells and mediators in bronchoalveolar lavage fluid from asthmatic and normal children. Methods: Children were recruited from elective surgical lists and a non-bronchoscopic bronchoalveolar lavage (BAL) was performed after induction of anaesthesia. Exhaled nitric oxide (parts per billion) was measured by two techniques: tidal breathing and restricted breath. Results: Median (interquartile range) exhaled nitric oxide measured by restricted breath was increased in asthmatics compared with normal children (24.3 (10.5–66.5) v 9.7 (6.5–16.5), difference between medians 14.6 (95% CI 5.1 to 29.9), p=0.001). In asthmatic children exhaled nitric oxide correlated significantly with percentage eosinophils (r=0.78, p<0.001 (tidal breathing) and r=0.78, p<0.001 (restricted breath)) and with eosinophilic cationic protein (r=0.53, p<0.01 (restricted breath)), but not with other inflammatory cells in the BAL fluid. The area under the receiver operator characteristic curves for the prediction of the presence of eosinophilic airways inflammation by exhaled nitric oxide (tidal and restricted) was 0.80 and 0.87, respectively. Conclusions: Exhaled nitric oxide correlates closely with percentage eosinophils in BAL fluid in asthmatic children and is therefore likely to be a useful non-invasive marker of airway inflammation.

Outgrown asthma does not mean no airways inflammation

Although some asthmatic children seem to recover from their asthma, 30–80% develop asthma again in later life. The underlying risk factors are unknown. The hypothesis for this study was that children with apparently outgrown asthma would have underlying airway inflammation. Nonbronchoscopic bronchoalveolar lavage was performed on normal children (n=35) and children who had wheezed previously (n=35). Eosinophils were raised in the lavage fluid of atopic children who had apparently outgrown asthma (median (interquartile range) 0.36 (0.05–0.74) compared to controls 0.10 (0–0.18), p=0.002). There was no relationship between length of remission and degree of airways eosinophilia. Thus, there is persistent airways inflammation in some children with outgrown asthma and this may be a risk factor for future relapse.

Inflammatory mediators in bronchoalveolar lavage samples from children with and without asthma

We investigated whether eosinophils and mast cells, found in the airways of children with wheeze, were activated during relatively asymptomatic periods.

Antioxidants and oxidative stress in BAL fluid of atopic asthmatic children

Earlier studies in adults have indicated that increased oxidative stress may occur in the blood and airways of asthmatic subjects. Therefore the aim of this study was to compare the concentrations of antioxidants and protein carbonyls in bronchoalveolar lavage fluid of clinically stable atopic asthmatic children (AA, n = 78) with our recently published reference intervals for nonasthmatic children (C, n = 124). Additionally, lipid peroxidation products (malondialdehyde) in bronchoalveolar lavage fluid and several antioxidants in plasma were determined. Bronchoalveolar lavage concentrations (median and interquartile range) of ascorbate [AA: 0.433 (0.294–0.678) versus C: 0.418 (0.253-0.646) μmol/L], urate [AA: 0.585 (0.412–0.996) versus C: 0.511 (0.372–0.687) μmol/L], α-tocopherol [AA: 0.025 (0.014–0.031) versus C: 0.017 (0.017–0.260) μmol/L], and oxidized proteins as reflected by protein carbonyls [AA: 1.222 (0.970–1.635) versus C: 1.243 (0.813–1.685) nmol/mg protein] were similar in both groups (p > 0.05 in all cases). The concentration of protein carbonyls correlated significantly with the number of eosinophils, mast cells, and macrophages in AA children only. Concentrations of oxidized proteins and lipid peroxidation products (malondialdehyde) correlated significantly in AA children (r = 0.614, n = 11, p = 0.044). Serum concentrations of ascorbate, urate, retinol, α-tocopherol, β-carotene, and lycopene were similar in both groups whereas α-carotene was significantly reduced in asthmatics. Overall, increased bronchoalveolar lavage eosinophils indicate ongoing airway inflammation, which may increase oxidatively modified proteins as reflected by increased protein carbonyl concentrations.

Possible association between passive smoking and lower exhaled nitric oxide in asthmatic children

In adults, both active and passive smoking reduce levels of exhaled nitric oxide (eNO); however, to date, passive exposure to environmental tobacco smoke (ETS) has not been shown to affect eNO in children. The authors recruited 174 asthmatic children (96 male, 78 female) and 79 nonasthmatic controls (46 male, 33 female) from a group of children aged 5 to 14 yr who attended a childrens hospital for an outpatient visit or elective surgery. Each subjects exposure to ETS was ascertained by questionnaire, and their eNO levels were measured. Asthmatic children had higher eNO levels (ppb) than nonasthmatic children (p = 0.04), and asthmatic children exposed to ETS had significantly lower eNO levels than unexposed children (p = 0.005). Exposure to ETS did not alter eNO levels in nonasthmatic children (p = 0.4). Results of the study suggest that ETS exposure is associated with lower eNO levels among childhood asthmatics. Consequently, ETS exposure may need to be considered when physicians interpret eNO levels in asthmatic children. Further study of the effects of ETS on eNO levels is recommended.

The repeatability of nonbronchoscopic bronchoalveolar lavage differential cell counts

Airway inflammation in children can be assessed by nonbronchoscopic bronchoalveolar lavage (BAL). Little is known about the repeatability of cell counts in the BAL obtained. Children (n=43) attending for elective surgery were studied. Cell counts were obtained following a nonbronchoscopic lavage. Two samples were obtained with either: 1) the catheter wedged in the same position (n=21) or 2) the catheter reinserted and wedged again (n=22). Slides (n=30) from nonbronchoscopic lavage samples were selected at random and two independent observers counted 500 cells on each slide on two occasions. The repeatability of the lavage sampling and cell counting was assessed for different cell types. The inter- and intra-observer repeatability for the differential cell counting demonstrated that there was good repeatability for all cell types except lymphocytes (interobserver: Lins concordance coefficient 0.42; repeatability coefficient 0.66). Quantification of eosinophil (%) was highly repeatable using either method (Lins concordance coefficient 1) 0.99, 2) 0.95; repeatability coefficient 1) 0.58, 2) 1.36). Nonbronchoscopic lavage is a repeatable technique for the quantification of eosinophils. Variation in the sampling method can be reduced by taking two separate samples and averaging the differential cell counts. Furthermore, increasing the number of cells counted should ensure accurate quantification of lymphocytes.

Antioxidants and protein carbonyls in bronchoalveolar lavage fluid of children: normal data

Antioxidant-oxidant imbalances in bronchoalveolar lavage fluid (BAL) are thought to contribute to oxidative stress in respiratory disease. However, normal reference ranges for BAL antioxidants and oxidized proteins in children are not available. In this study, we recruited 124 children attending for elective surgery for a noninflammatory condition; 83 were nonasthmatic, nonatopic (N) and 41 were nonasthmatic, atopic (NA). A nonbronchoscopic lavage was performed and ascorbate, uric acid, α-tocopherol, and protein carbonyl (as a measure of oxidative damage) concentrations were determined in BAL fluid. The 95% reference range was 0.112–1.897 μmol/L for ascorbate, 0.149–2.163 μmol/L for urate, 0.0029–0.066 μmol/L for α-tocopherol, and 0.280–4.529 nmol/mg for protein carbonyls in BAL fluid. Age, gender, and exposure to environmental tobacco smoke did not affect the concentration of ascorbate, urate, α-tocopherol, or protein carbonyls. However, in multiple linear regression analyses, the type of home heating (glass-fronted fires or oil-fired central heating) was found to influence ascorbate and urate concentrations in the BAL fluid (β-coefficient for ascorbate: 0.445, p = 0.031; for urate: 0.114, p = 0.001). There was no significant difference between the N and NA group in BAL fluid concentrations of ascorbate, urate, or protein carbonyls. The α-tocopherol concentration was significantly increased in the NA group (p = 0.037). Uric acid and α-tocopherol concentrations in BAL fluid and serum were not correlated. Intriguingly, serum and BAL ascorbate concentrations were significantly correlated (r = 0.297, p = 0.018, n = 63), which may offer an explanation for why supplementing the diet with vitamin C can improve asthma symptoms. Further studies will investigate the role of BAL antioxidant concentrations in children with inflammatory respiratory diseases.

Exhaled nitric oxide in relation to the clinical features of childhood asthma

Background. Exhaled nitric oxide (ENO) has been shown to be a noninvasive marker of eosinophilic inflammation in asthmatic children. Few studies have evaluated the relationship between ENO levels and the clinical features of children with asthma. The aim of this study was to examine children attending a routine asthma clinic and evaluate the relationship between ENO levels and clinical parameters including decision making. Methods. Asthmatic children (n = 133, aged 5 to 14 years) attending a hospital asthma clinic were studied. ENO levels were measured and compared between subgroups of subjects according to recent symptoms, asthma control and treatment, and the clinicians decision (blinded to ENO levels) regarding further management. Results. ENO levels (median [IQR] ppb) were significantly elevated in children who had recent symptoms compared to those without recent symptoms (14.6 [6.5 to 45.3] vs. 6.0 [3.2 to 17.4], difference between medians 8.6, 95% confidence interval [CI] (1.8 to 13.9, p = 0.004). ENO levels differed significantly between the controlled and uncontrolled subgroups (8.5 [4.2 to 26.4] vs. 26.4 [5.0 to 62.0], difference between medians 17.9, 95% CI 0.1 to 22.8, p = 0.03) and between the three treatment decision subgroups (up, down, or unchanged; p < 0.001). Conclusions. ENO levels are strongly related to the clinical features of childhood asthma and the clinical decision making process. To fully evaluate the role of ENO in the clinical management of asthma, this “proof of concept” study paves the way for prospective randomized trials of the inclusion of ENO levels in the decision making process in childhood asthma.

Serum eosinophil cationic protein (ECP): reference values in healthy nonatopic children

Background: Although serum ECP concentrations have been reported in normal children, there are currently no published upper cutoff reference limits for serum ECP in normal, nonatopic, nonasthmatic children aged 1–15 years.

Interleukin-4 and interleukin-4 soluble receptor α levels in bronchoalveolar lavage from children with asthma

BACKGROUND In asthma there is increased expression of the Th2-type cytokine interleukin-4 (IL-4). IL-4 is important in immunoglobulin isotype switching to immunoglobulin E and adhesion of eosinophils to endothelium. OBJECTIVE We hypothesized that levels of IL-4 in bronchoalveolar lavage (BAL) fluid would be increased in stable, atopic asthmatic children compared with controls and that levels of its physiologic inhibitor IL-4 soluble receptor alpha (IL-4sR alpha) would be correspondingly decreased. METHODS One hundred sixteen children attending a childrens hospital for elective surgery were recruited. A nonbronchoscopic BAL was performed, and IL-4 and IL-4sR alpha were measured in the BAL supernatants. RESULTS There was no significant difference in IL-4 concentrations between atopic asthmatic children, atopic normal controls, and nonatopic normal controls [0.13 pg/mL (0.13 to 0.87) vs 0.13 pg/mL (0.13 to 0.41) vs 0.13 pg/mL (0.13 to 0.5), P = 0.65]. IL-4sR alpha levels were significantly increased in asthmatic patients compared with atopic controls [6.4 pg/mL (5.0 to 25.5) vs 5.0 pg/mL (5.0 to 9.9), P = 0.018], but not when compared with the nonatopic controls [5.2 pg/mL (5.0 to 10.6), P = 0.19]. CONCLUSIONS Contrary to expectation, IL-4sR alpha levels are increased in BAL from stable asthmatic children compared with nonatopic controls, and we speculate that IL-4sR alpha is released by inflammatory cells in the airways to limit the proinflammatory effects of IL-4.