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American Journal of Respiratory and Critical Care Medicine Vol 166. pp. 301-306, (2002)
© 2002 American Thoracic Society

Original Article

Increased Exhaled Cysteinyl-Leukotrienes and 8-Isoprostane in Aspirin-induced Asthma

Adam Antczak, Paolo Montuschi, Sergei Kharitonov, Pawel Gorski and Peter J. Barnes

Department of Thoracic Medicine, National Heart and Lung Institute, Imperial College, London, United Kingdom; and Department of Pneumology and Allergology, Medical University of Lodz, Lodz, Poland

Correspondence and requests for reprints should be addressed to Adam Antczak, M.D., Department of Pneumology and Allergology, Medical University of Lodz, Kopcinskiego Str. 22, 50-153 Lodz, Poland. E-mail: magant{at}kki.net.pl


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
The pathogenesis of aspirin-induced asthma (AIA) has not yet been clearly elucidated, although eicosanoid metabolites appear to play an important role. We hypothesized that levels of eicosanoids in exhaled air condensate are abnormal in patients with AIA and that they change in patients receiving steroid therapy. We measured cysteinyl-leukotrienes (cys-LTs), prostaglandin E2 (PGE2), and leukotriene B4 (LTB4), and also 8-isoprostane as a marker of oxidative stress, by enzyme immunoassay in exhaled breath condensate from patients with AIA (17 steroid naive; mean age, 41 ± 23 years; FEV1, 63%pred), 26 patients with aspirin-tolerant asthma (ATA) (11 steroid naive; mean age, 47 ± 18 years; FEV1, 69%pred), and 16 healthy subjects (mean age, 45 ± 17 years; FEV1, 93%pred). Cys-LTs were significantly higher in steroid-naive patients with AIA compared with steroid-naive patients with ATA and healthy subjects (152.3 ± 30.4 and 36.6 ± 7.1 versus 19.4 ± 2.8 pg/ml; p < 0.05 and p < 0.05, respectively). Steroid-naive patients with AIA also had higher levels of 8-isoprostane than normal subjects (131.8 ± 31.0 versus 21.9 ± 4.5 pg/ml; p < 0.05). There were significantly lower levels of both cys-LTs and 8-isoprostanes in steroid-treated patients with AIA. There was no difference in either the PGE2 or LTB4 level between the patient groups. This is the first study to show that cys-LTs and 8-isoprostanes are elevated in expired breath condensate of steroid-naive patients with AIA, and that cys-LTs are decreased in steroid-treated patients. Exhaled PGE2 levels are not reduced, so that it is unlikely that a deficiency of PGE2 is an important mechanism, whereas exhaled LTB4 levels are unchanged, indicating an abnormality beyond 5-lipoxygenase.

Key Words: aspirin-induced asthma • cysteinyl-leukotrienes • exhaled breath condensate • 8-isoprostane • prostaglandin E2


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Aspirin-induced asthma (AIA) is a distinct clinical syndrome observed in 5–15% of adult patients with asthma, and almost 2.5 times more often in women (1, 2). In patients with AIA, ingestion of aspirin and other nonsteroidal antiinflammatory drugs leads to an acute asthma attack, often accompanied by rhinorrhea, conjunctival irritation, and flushing of the head and neck (3). All the drugs precipitating asthmatic attacks are inhibitors of cyclooxygenase (4). From the clinical point of view, AIA is usually more severe, has a later onset than allergic asthma, and is associated with nasal polyposis or sinusitis (3). Persistent inflammation in the airways of all subjects with asthma is characterized by eosinophilic infiltration, desquamation of epithelium, and increased thickness of the reticular layer of the basement membrane. Bronchial biopsy specimens from patients with AIA show a 4-fold increase in eosinophils compared with those who tolerate aspirin (ATA) (5). Because the frequency of atopy is similar in patients with AIA and in patients with ATA (6), the increased influx of eosinophils into the airways of patients with AIA is likely to result from a nonatopic mechanism. Chronic viral infection and/or autoimmunization are putative causes of AIA (7). The presence of human leukocyte antigen DPB1 (HLA-DPB1) association suggests that immune recognition of an unknown antigen may also be involved in the etiology of AIA (8). Activated eosinophils and mast cells are capable of producing several inflammatory mediators, including cysteinyl-leukotrienes (cys-LTs) (LTC4, LTD4, LTE4), which are potent bronchoconstrictors and are likely to play an important role in AIA. LTC4 synthase, the enzyme controlling cys-LT synthesis, is expressed in blood eosinophils of patients with asthma (9). Urinary excretion of LTE4 is increased by 2–10 times in patients with AIA, compared with patients with ATA, providing evidence of constitutive chronic activation of the 5-lipoxygenase/LTC4 synthase pathway (10). After aspirin challenge, a marked increase in production of cys-LTs is observed in the bronchoalveolar lavage fluid (BALF) and/or urine of patients with AIA, but not in those with ATA (10, 11). 5'-Lipoxygenase inhibitors and cys-LT receptor antagonists markedly attenuate aspirin-induced respiratory reactions (12–15). The overexpression of LTC4 synthase in bronchial biopsy specimens of patients with AIA is correlated with aspirin sensitivity and is fivefold higher than in subjects with ATA (5). Moreover, baseline concentrations of cys-LTs measured in BALF of patients with AIA correlate positively with counts of inflammatory cells (predominantly eosinophils) immunoreactive for LTC4 synthase in the bronchial mucosa (5). Because immunohistochemical differences between patients with AIA and those with ATA are several-fold different for LTC4 synthase and interleukin-5–positive cells, a polymorphism directed to the regulation of LTC4 expression was proposed as a predisposing factor in AIA (5). The LTC4 synthase gene is located on the long arm of chromosome 5 and is adjacent to a region of crucial candidate genes for asthma (cluster of cytokine genes involved in the allergic phenotype) and this region of chromosome 5 has been implicated in asthma by linkage analysis (16). In fact, a genetic predisposition to cys-LT pathway upregulation can be related to overactive expression of the LTC4 synthase allele. Sanak has shown (9) that the gene encoding LTC4 synthase exists as two common alleles, distinguished by an adenine-to-cytosine conversion at a site 444 base pairs upstream of the translation start site. This single-nucleotide polymorphism affects binding of transcription factors and influences the transcription rate, predisposing to AIA. Moreover, the –444C allelic frequency is significantly higher in patients with AIA as compared with matched patients with ATA and healthy control subjects. Patients with AIA also have upregulated LTC4 synthase messenger RNA expression in peripheral blood eosinophils (9).

Szczeklik and coworkers have also shown diminished PGE2 levels in BALF obtained from patients with AIA, which were further decreased after administration of aspirin (17). PGE2 relaxes airway smooth muscles and exerts potent antiinflammatory activity (18). Moreover, inhaled PGE2 can inhibit aspirin-induced bronchoconstriction and urinary LTE4 excretion (19). It is postulated that the failure in a PGE2-braking mechanism with increased sensitivity to inhibition by nonsteroidal antiinflammatory drugs is responsible for overproduction of cys-LTs in patients with AIA.

Several studies have highlighted differences between the effects of corticosteroids on eicosanoid production (especially LTs) in vivo and in vitro. Baseline excretion of cys-LTs is not suppressed by steroids (20). Inhaled fluticasone propionate, which effectively prevents asthma attacks, has no effect on increased urinary LTE4 excretion after allergen challenge (21). On the other hand, steroids are potent inhibitors of cys-LT production by peripheral blood mixed leukocytes (22) and diminish the influx of eosinophil to the bronchial mucosa (23). On the basis of these data, it would be interesting to evaluate whether steroid treatment can diminish exhaled eicosanoids in AIA.

Cys-LTs and PGE2 are eicosanoids that have been implicated in the development of AIA. Leukotriene B4 (LTB4) plays an important role in neutrophil recruitment and activation at the site of inflammation (24). 8-Isoprostane is a biomarker of oxidative stress in breath condensate in inflammatory airway diseases such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis (25–27).

The aim of this study was to determine whether cys-LTs, PGE2, LTB4, and 8-isoprostane levels are increased in expired breath condensate in patients with AIA compared with patients with ATA and healthy subjects and whether there are any differences between steroid-naive and steroid-treated patients with AIA. Exhaled breath condensate has the advantage of being noninvasive and also directly samples mediators from the respiratory tract, thus giving a more direct approach to gaining insight about inflammatory mediators in asthma.


   METHODS
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Study Population
We included 31 patients with AIA (17 steroid-naive; mean age, 41 ± 23 years; 23 female; FEV1, 63%pred), 26 aspirin-tolerant patients with mild to moderate asthma (ATA) (11 steroid-naive; mean age, 47 ± 18 years; 15 female; FEV1, 69%pred), and 16 healthy subjects (mean age, 45 ± 17 years; FEV1, 93%pred; 7 female), who volunteered for this study (Table 1) . Normal subjects had no history of any respiratory symptoms or respiratory infection. Asthma was diagnosed by history of recurrent wheezing and chest tightness and a bronchodilator response of more than a 15% increase in FEV1 with inhaled albuterol (200 µg) and the presence of airway hyperreactivity after a histamine challenge test with PC20 of less than 8 mg ml-1 (28). Patients with ATA reported use of nonsteroidal antiinflammatory drugs with no adverse effects. The diagnosis of aspirin intolerance was confirmed by aspirin provocation test (29) performed during the 2 years preceding the study. Briefly, aspirin provocation was performed with a nebulizer and crystalline lysine aspirin (Aspirol; Bayer, Levercusen, Germany). The provocation was stopped when the FEV1 dropped by 20% from baseline values or when the maximal dose of aspirin (115.2 mg) had been reached.


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  		TABLE 1. Characteristics of study population

 
All patients were lifelong nonsmokers, and those with asthma were stable with no changes in asthma symptoms and medication for at least 1 month. Accepted treatment included short-acting inhaled ß2-agonists and steroids (inhaled budesonide, not more than 1,200 µg/day; or oral methyloprednisolone, not more than 8 mg/day). All subjects gave informed written consent, and the study was approved by the ethics committee of the Medical University of Lodz (Lodz, Poland).

Measurement of Exhaled Eicosanoids
The exhaled breath condensate was collected in a tube installed in a polystyrene foam container filled with dry ice. The collecting part of the tube (170 cm of a total of 230 cm) was covered with dry ice. The temperature in the tube vicinity, measured with a thermocouple, ranged from -43 to -35°C and allowed condensation of the exhaled air. Patients were asked to breathe out spontaneously for 10–15 minutes through a mouthpiece with a saliva trap connected to the tube. The respiratory rate ranged from 15 to 20 breaths/minute. Subjects wore a noseclip and rinsed their mouths with distilled water just before and after 7 minutes of cooling to reduce evaporation of eicosanoids from saliva and nasal spaces. At the end of collection, the tube was carefully removed from the container, and 1- to 2.5-ml aliquots of condensate were transferred to Eppendorf tubes. Samples were stored at -80°C for no longer than 4 weeks until measurements (30).

Measurement of Immunoreactive Leukotrienes
The cys-LT concentration in breath condensate was measured with a specific enzyme immunoassay (EIA) kit (Cayman Chemical, Ann Arbor, MI) as previously described (31). LTB4 in breath condensate was measured with an EIA kit (Cayman Chemical). The antiserum used in this assay has a cross-reactivity of 100% with LTB4, 39% with 6-trans-LTB4, and < 0.01% each with LTC4, LTE4, LTD4, and LTF4. The detection limit is 13 pg/ml for cys-LTs and 4.43 pg/ml for LTB4.

Measurement of Immunoreactive 8-Isoprostane
The 8-isoprostane concentration in breath condensate was measured with an EIA kit (Cayman Chemical) as previously described (25). The detection limit was 5 pg/ml.

Measurement of Immunoreactive PGE2
The PGE2 concentration in breath condensate was measured with a specific EIA kit (Cayman Chemical). The antiserum used in this assay has a cross-reactivity of 100% with PGE2, 43% with PGE3, 18.7% with PGE1, and 0.1% each with PGF2{alpha}, PGA1, and PGA2. The detection limit at 4° C is 15 pg/ml.

Statistical Analysis
The nonparametric Kruskal–Wallis test for multiple comparisons was used to compare groups. Levels of measured mediators below the detection limit were arbitrarily assumed to be 0.5 of the detection limit. Linear regression analysis was used to assess the relationship between measured parameters. All data are expressed as means ± standard error of the mean and significance was defined as a p value < 0.05.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Cysteinyl-leukotrienes
There were higher concentrations of cys-LTs in patients with AIA compared with those with ATA, and in both asthmatic groups, levels were significantly higher than in normal subjects (108.4 ± 19.2 and 59.0 ± 7.7 versus 19.4 ± 2.7 pg/ml; p < 0.001 and p < 0.05, respectively) (Table 2) . Figure 1 shows mean levels of cys-LTs in patients with AIA and patients with ATA, divided into steroid-treated and steroid-naive subsets and healthy subjects. Steroid-naive patients with AIA had significantly higher levels of cys-LTs in expired breath condensate than did steroid-naive subjects with ATA. Moreover, both steroid-naive AIA and ATA groups had higher levels of cys-LTs than healthy subjects (152.3 ± 30.4 and 50.7 ± 15.7 versus 19.4 ± 2.8 pg/ml; p < 0.05 and p < 0.05, respectively). Steroid-treated patients with AIA had higher levels of cys-LTs than normal control subjects and significantly higher levels than ATA steroid-treated patients (55.2 ± 10.1 versus 19.4 ± 2.8 and 36.6 ± 7.1 pg/ml; p < 0.05 and p < 0.05, respectively). Steroid treatment is associated with lower levels of cys-LTs in patients with AIA but not in patients with ATA (55.2 ± 10.1 versus 152.3 ± 30.3 and 36.6 ± 7.1 versus 50.7 ± 15.7 pg/ml; p < 0.05 and p > 0.05, respectively).


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  		TABLE 2. Eicosanoids in expired breath condensate of patients with aspirin-intolerant asthma and patients with aspirin-tolerant asthma, calculated for whole groups*

 


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  		Figure 1. Cys-LTs in expired breath condensate of patients with asthma. AIA S = steroid-treated patients with AIA; AIA S-N = steroid-naive patients with AIA; ATA S = steroid-treated patients with ATA; ATA S-N = steroid-naive patients with ATA. Detection limit, 13 pg/ml.

 
Leukotriene B4
There were no differences between LTB4 concentrations in expired breath condensate from AIA and ATA groups compared with normal subjects (Figure 2) .



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  		Figure 2. LTB4 in expired breath condensate of patients with asthma. AIA S = steroid-treated patients with AIA; AIA S-N = steroid-naive patients with AIA; ATA S = steroid-treated patients with ATA; ATA S-N = steroid-naive patients with ATA. Detection limit, 4.43 pg/ml.

 
Prostaglandin E2
No differences were found between steroid-treated and steroid-naive AIA and ATA groups in terms of PGE2 levels in expired breath condensate (Table 2 and Figure 3) . However, steroid-naive patients with AIA had almost 50% higher concentrations of PGE2 than did healthy subjects and almost double that of matched ATA patients, but because of the high variability the difference did not achieve statistical significance.



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  		Figure 3. PGE2 levels in expired breath condensate of patients with asthma. AIA S = steroid-treated patients with AIA; AIA S-N = steroid-naive patients with AIA; ATA S = steroid-treated patients with ATA; ATA S-N = steroid-naive patients with ATA. Detection limit, 15 pg/ml.

 
8-Isoprostane
Although there is no difference in 8-isoprostane levels between patients with AIA and patients with ATA (Table 2), both AIA and ATA groups had higher 8-isoprostane concentrations in expired breath condensate compared with normal control subjects (90.0 ± 19.0 and 79.2 ± 19.5 compared with 21.9 ± 4.5 pg/ml; p < 0.05). When patients with asthma were divided into steroid-treated and steroid-naive subgroups, steroid-naive AIA and ATA groups had greater 8-isoprostane levels compared with healthy subjects (131.8 ± 31.0 and 77.3 ± 21.9 compared with 21.9 ± 4.5 pg/ml; p < 0.05 and p < 0.05, respectively) (Figure 4) . There was no significant difference in 8-isoprostane levels between steroid-naive AIA and ATA groups, but there was a difference in 8-isoprostane levels between steroid-treated and steroid-naive AIA groups (39.3 ± 7.4 versus 131.5 ± 31.0 pg/ml; p < 0.05). There was no difference between steroid-treated AIA and ATA groups, but levels of 8-isoprostane were higher in both groups compared with healthy control subjects (39.3 ± 7.4 and 80.6 ± 30.5 compared with 21.9 ± 4.5 pg/ml; p > 0.05 and p < 0.05, respectively).



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  		Figure 4. 8-Isoprostane in expired breath condensate of patients with asthma. AIA S = steroid-teated patients with AIA; AIA S-N = steroid-naive patients with AIA; ATA S = steroid-treated patients with ATA; ATA S-N = steroid-naive patients with ATA. Detection limit, 5 pg/ml.

 
Correlations
No correlation was found between any eicosanoid level and pulmonary function or age in any group of patients.


   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
In this study, we have measured cys-LTs, LTB4, PGE2, and 8-isoprostanes in expired breath condensate from patients with AIA and patients with ATA and compared them with healthy control subjects. We found that cys-LTs were significantly increased in patients with AIA compared with those with ATA and were higher in both asthmatic groups compared with healthy subjects. Increased levels of 8-isoprostanes in exhaled breath condensate were found both in patients with AIA and patients with ATA compared with normal subjects, whereas LTB4 and PGE2 levels were not increased in either group.

Expired breath condensate provides a new, noninvasive, and easily performed means of looking at the local inflammatory process in the airways, without the need to undertake invasive procedures such as bronchoscopy. It is well tolerated by patients and no adverse events have been reported. This technique has never been used to study subjects with AIA; however, a growing body of evidence suggests that it is a useful way to monitor markers of inflammation and oxidative stress in various respiratory tract diseases, such as asthma, chronic obstructive pulmonary disease, and cryptogenic fibrosing alveolitis (25–27).

An altered metabolism of arachidonic acid is believed to be the underlying mechanism of bronchoconstriction in patients with AIA (1). Increased cys-LT levels in exhaled breath condensate of patients with AIA and patients with ATA compared with healthy control subjects confirm previous measurements of high urinary cys-LTs reflecting increased leukotriene production (32). This is supported by studies of bronchial and nasal lavage fluids, which also show increased levels of cys-LTs (33). It is also consistent with the results of Schafer and coworkers (34), who showed that the profile of eicosanoid mediators is changed in AIA not only after, but also before, aspirin challenge. In comparing basal release of eicosanoids from peripheral blood cells before challenge, cys-LTs were found to be significantly elevated and PGE2 was reduced in patients with AIA, whereas patients with ATA had elevated basal cys-LTs but only slightly reduced basal PGE2 levels. After aspirin challenge, there was a marked increase in the already elevated cys-LTs in patients with AIA, but not in subjects with ATA (34). Our study now demonstrates local overproduction of cys-LTs in the airways in AIA and is consistent with a study showing that peripheral blood eosinophils obtained from all patients with asthma have increased expression of mRNA encoding LTC4 synthase, and that this mRNA is particularly increased in eosinophils obtained from patients with AIA (9). Eosinophils are increased to a greater extent in the bronchial mucosa in AIA compared with ATA, and this supports the view that overexpression of LTC4 synthase may play a pathogenic role in AIA. Interestingly, the levels of exhaled cys-LTs were lower in those AIA subjects who received inhaled steroid therapy. Plasma levels of LTE4 are reduced after oral prednisolone in acute exacerbations of chronic obstructive pulmonary disease (35). Oral prednisone reduces symptoms and inhaler use, but has no significant effect on BALF eicosanoid levels in volunteers with atopic asthma at baseline and after allergen challenge. It could, however, reduce the synthesis of eicosanoids in macrophage-rich BALF cells in vitro (36). In our study, there was a reduction in exhaled cys-LTs in steroid-treated compared with steroid-naive patients with AIA. This suggests that inhaled steroid therapy suppresses local production of cys-LTs in the airways, and thus exerts an antiinflammatory effect in AIA. Corticosteroids eliminate eicosanoid output from human airway epithelial cells in vitro. Inhaled steroids can also suppress the influx of eosinophils, and thus lead to a reduction in synthesis of leukotrienes in vitro (37).

In contrast to the more elevated baseline cys-LTs, PGE2 levels in BALF have been reported to be lower in patients with AIA than in patients with ATA. After bronchoscopic instillation of L-lysine aspirin, there is a significant reduction of PGE2 to undetectable levels, an effect consistent with inhibition of cyclooxygenase enzyme by aspirin (17). Nasal polyp epithelial cells cultured from patients with AIA generate significantly less PGE2, both spontaneously and after stimulation with calcium ionophore, than do cells from patients with ATA (38). This is consistent with the results of Picado and coworkers, who found reduced expression of cyclooxygenase-2 mRNA in nasal polyps from patients with AIA (39). However, these results were not confirmed in the bronchial mucosa of subjects with AIA. It seems plausible that a deficiency in PGE2 production may play a role in AIA. Assuming that PGE2 possesses bronchoprotective activity, low levels may be unable to confront the bronchoconstrictive effects of cys-LTs. Moreover, PGE2 may also inhibit the release of LTB4 from neutrophils in a concentration-dependent manner—an effect of potential importance in difficult asthma (40). Our results show that local levels of PGE2 in the airways are not reduced in AIA compared with ATA and healthy subjects.

8-Isoprostane, a stable prostaglandin-like arachidonate product formed on membrane phospholipids by the action of reactive oxygen species, is postulated to be a reliable in vivo biomarker of lipid peroxidation (41, 42). 8-Isoprostane in breath condensate appears to reflect oxidative stress and is progressively increased with the severity of asthma (25). In this study, we observed higher concentrations of 8-isoprostane in all groups compared with studies done previously (25), but we used a different type of condenser and a lower temperature to obtain samples. In this study, we found increased levels of 8-isoprostane in expired breath condensate in steroid-naive patients with AIA compared with steroid-treated patients with AIA and healthy control subjects. There were no differences between ATA subgroups. Steroid-naive patients with ATA had almost three times higher levels of 8-isoprostane than healthy control subjects, but because of the large variability in the ATA group, there was no significant difference. Eosinophil numbers are greater in the bronchial mucosa of subjects with AIA compared with patients with ATA. Cells (including eosinophils) recovered from the BALF and blood of subjects with asthma generate greater amounts of oxidants than those of normal subjects (43). The oxidative stress produced by eosinophils may be important, as these cells possess a high capacity to generate reactive oxygen species (44). It is likely that the intensity of inflammation, as reflected by oxidative stress in the airways of patients with AIA, is greater than in subjects with ATA. Increased levels of 8-isoprostane in the breath condensate of patients with mild ATA appear to be independent of steroid treatment, and this is confirmed in our study (25). However, our results indicate that steroid treatment decreases levels of 8-isoprostane in exhaled breath condensate of subjects with AIA, suggesting that there is a steroid-dependent inhibitory mechanism in AIA that is not present in ATA. In vitro 8-isoprostane production is decreased by dexamethasone via cyclooxygenase-2 inhibition (45). Inhaled beclomethasone significantly prevents allergen-induced synthesis of 8-isoprostane in subjects with atopic asthma, as measured by 8-isoprostane content in both urine and BALF (41). Abrogation of oxidant stress and lipid peroxidation in the allergen-induced asthmatic reaction by steroids may be the result of a reduction in the number and activation of cells producing reactive oxygen species. Inhaled steroids are able to diminish the influx of eosinophils and release of inflammatory mediators (41). Interestingly, in patients with ATA, inhaled steroids are able to decrease oxidant stress as reflected by hydrogen peroxide in expired breath condensate (46).

We did not find any differences in LTB4 levels between AIA and ATA groups, irrespectively of steroid therapy. LTB4 is a potent chemoattractant and activator of neutrophils (47). There is no evidence that neutrophilic infiltration is increased in AIA, so it is not surprising that exhaled LTB4 levels are not elevated. The fact that LTB4 levels in exhaled condensate are not elevated, whereas cys-LTs are increased, also indicates that there is no increase in phospholipase A2 or 5-lipoxygenase enzyme activity in patients with AIA, but that the abnormality is downstream of these enzymes, lending further support to the view that LTC4 synthase activity is abnormal in AIA.

This is the first study to measure eicosanoids in expired breath condensate in AIA. We have shown that cys-LT and 8-isoprostane concentrations are elevated in expired air condensate of steroid-naive patients with AIA and patients with ATA, and that steroid treatment leads to a significant decrease in these compounds in patients with AIA but not ATA. By contrast, exhaled LTB4 and PGE2 levels are not reduced, and therefore, these are unlikely to be an important mechanism.


   Acknowledgments
 
A. A. is the recipient of a Research Fellowship from the Count Potocki Foundation (Poland); P. M. is the recipient of a Research Fellowship from the National Research Council of Italy.

Received in original form January 5, 2001; accepted in final form April 8, 2002


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

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