Late-phase Airway Obstruction Research Articles

Bronchial Allergen Challenge of Patients with Atopic Asthma Triggers an Alarmin (IL-33, TSLP, and IL-25) Response in the Airways Epithelium and Submucosa

The alarmin cytokines IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) play a critical role in asthma pathogenesis by inducing mucosal Th2-type cytokine production. Although environmental exposure to aeroallergens has been proposed as an alarmin trigger in asthma, there has been no systematic parallel study of the effects of allergen exposure on the expression of these cytokines in the airways of human asthmatics. Using single and sequential double immunohistochemistry, we evaluated the numbers and phenotypes of IL-25-, IL-33-, and TSLP-immunoreactive cells in sections of bronchial biopsies from mild atopic asthmatics (n = 16) before and 24 h after allergen inhalational challenge. Allergen challenge highly increased expression of baseline immunoreactivity for IL-25, IL-33, and TSLP, both in the bronchial epithelium and submucosa (p < 0.001), to a degree that correlated with the extent of the late phase of airway obstruction. Aside from epithelial cells, the principal source of immunoreactivity for all three alarmins, TSLP, and IL-33 immunoreactivity colocalized principally with endothelial cells and mast cells, neutrophils, and fibroblasts, whereas IL-25 immunoreactivity colocalized principally with eosinophils as well as endothelial cells, mast cells, and fibroblasts. The data implicate that allergen challenge directly increases airway alarmin expression in atopic asthmatics to a degree correlating with increase late-phase airway obstruction, affirming these molecules as potential molecular targets for the inhibition of allergen-induced airway inflammation and obstruction.

IL-17A Promotes the Exacerbation of IL-33–Induced Airway Hyperresponsiveness by Enhancing Neutrophilic Inflammation via CXCR2 Signaling in Mice

Neutrophilic airway inflammation is a hallmark of patients with severe asthma. Although we have reported that both IL-33 and IL-17A contributed to IgE-mediated neutrophilic inflammation in mice, the relationship remains unclear. In this article, we examined how IL-17A modifies IL-33-induced neutrophilic inflammation and airway hyperresponsiveness (AHR). IL-33 was intratracheally administered to BALB/c mice on days 0-2; furthermore, on day 7, the effect of the combination of IL-33 and IL-17A was evaluated. Compared with IL-33 or IL-17A alone, the combination exacerbated neutrophilic inflammation and AHR, associated with more increased levels of lung glutamic acid-leucine-arginine(+) CXC chemokines, including CXCL1, CXCL2, and CXCL5, and infiltration by alveolar macrophages expressing CXCR2. Treatment with anti-CXCR2 mAb or depletion of alveolar macrophages repressed neutrophilic inflammation and AHR; in addition, depletion of neutrophils suppressed AHR. These findings prompted us to examine the role of CXCR2 in IgE-sensitized mice; a single treatment with anti-CXCR2 mAb in the seventh Ag challenge inhibited late-phase airway obstruction, AHR, and neutrophilic inflammation. In addition to inhibition, multiple treatments during the fourth to seventh challenge attenuated early-phase airway obstruction, eosinophilic inflammation, and goblet cell hyperplasia associated with the reduction of Th2 cytokine production, including IL-4, IL-5, and IL-13. Collectively, IL-33 cooperated with IL-17A to exacerbate AHR by enhancing neutrophilic inflammation via CXCR2 signaling; furthermore, CXCR2 signaling derived Th2 responses. We thus suggest the underlying mechanisms of IL-33 and IL-17A in allergic asthma and CXCR2 as potential therapeutic targets for the disease.

Roles of basophils and mast cells infiltrating the lung by multiple antigen challenges in asthmatic responses of mice

Mast cell hyperplasia has been observed in the lungs of mice with experimental asthma, but few reports have studied basophils. Here, we attempted to discriminate and quantify mast cells and basophils in the lungs in a murine asthma model, determine if both cells were increased by multiple antigen challenges and assess the roles of those cells in asthmatic responses. Sensitized Balb/c mice were intratracheally challenged with ovalbumin four times. Mast cells and basophils in enzymatically digested lung tissue were detected by flow cytometry. An anti-FcεRI monoclonal antibody, MAR-1, was i.p. administered during the multiple challenges. The numbers of both mast cells (IgE(+) C-kit(+) ) and basophils (IgE(+) C-kit(-) CD49b(+) ) increased in the lungs after three challenges. Treatment with MAR-1 completely abolished the increases; however, a late-phase increase in specific airway resistance (sRaw), and airway eosinophilia and neutrophilia were not affected by the treatment, although the early-phase increase in sRaw was suppressed. MAR-1 reduced antigen-induced airway IL-4 production. Basophils infiltrating the lung clearly produced IL-4 after antigen stimulation in vitro; however, histamine and murine mast cell protease 1 were not increased in the serum after the challenge, indicating that mast cell activation was not evoked. Both mast cells and basophils infiltrated the lungs by multiple intratracheal antigen challenges in sensitized mice. Neither mast cells nor basophils were involved in late-phase airway obstruction, although early-phase obstruction was mediated by basophils. Targeting basophils in asthma therapy may be useful for an early asthmatic response.

Cold air‐induced late‐phase bronchoconstriction in horses

Inspired air is warmed to body temperature and fully humidified by the upper airway mucosa under normal resting conditions. This conditioning process may not be completed by the upper airways during conditions of increased minute ventilation or when the inspired air is unusually cold, resulting in cooling and desiccation of lower respiratory surfaces. Excess heat and water loss from intrapulmonary airways is believed to be the provocative stimulus for exercise-induced bronchoconstriction (occurring immediately after exercise) and associated late phase airway obstruction (occurring a few hours after exercise). Exercise while breathing cold air results in airway obstruction in horses. Eight healthy horses performed a 15 min submaximal exercise challenge in a random crossover design. Independent variable was inspired air temperature during the challenge (25 or -5 degrees C). The dependent variables were total respiratory impedance, resistance, and reactance at 5, 24 and 48 h post exercise challenge, expressed as a percentage of the prechallenge baseline. No significant effect of inspired air temperature was found on any respiratory mechanical parameter 5 h after exercise challenge. However, cold inspired air was associated with higher respiratory impedance and resistance 48 h after the exercise challenges. These findings support the hypothesis that submaximal exercise while breathing subfreezing air can adversely affect respiratory mechanical properties in normal horses. However, the timecourse for development of abnormal respiratory mechanical properties is longer than that reported in other mammals. Exercise in cold weather may be a common cause of lower airway disease in horses.

Eicosanoids modulate hyperpnea-induced late phase airway obstruction and hyperreactivity in dogs

A canine model of exercise-induced asthma was used to test the hypothesis that the development of a late phase response to hyperventilation depends on the acute production of pro-inflammatory mediators. Peripheral airway resistance, reactivity to hypocapnia and aerosol histamine, and bronchoalveolar lavage fluid (BALF) cell and eicosanoid content were measured in dogs ∼5 h after dry air challenge (DAC). DAC resulted in late phase obstruction, hyperreactivity to histamine, and neutrophilic inflammation. Both cyclooxygenase and lipoxygenase inhibitors administered in separate experiments attenuated the late phase airway obstruction and hyperreactivity to histamine. Neither drug affected the late phase inflammation nor the concentrations of eicosanoids in the BALF obtained 5 h after DAC. This study confirms that hyperventilation of peripheral airways with unconditioned air causes late phase neutrophilia, airway obstruction, and hyperreactivity. The late phase changes in airway mechanics are related to the hyperventilation-induced release of both prostaglandins and leukotrienes, and appear to be independent of the late phase infiltration of inflammatory cells.

Heparin inhibits hyperventilation-induced late-phase hyperreactivity in dogs.

Inhalation of heparin attenuates hyperventilation-induced bronchoconstriction in humans and dogs. The purpose of this study was to determine whether heparin inhibits the late-phase response to hyperventilation, which is characterized by increased peripheral airway resistance (RP), eicosanoid mediator production, neutrophilic/ eosinophilic inflammation, and airway hyperreactivity (AHR) at 5 h after dry air challenge (DAC). Fiberoptic bronchoscopy was used to record RP and airway reactivity (DeltaRP) to aerosol and intravenous histamine before and 5 h after DAC. Bronchoalveolar lavage fluid (BALF) cells and eicosanoid mediators were also measured approximately 5 h after DAC. DAC of vehicle-treated bronchi resulted in late-phase airway obstruction (approximately 120% increase over baseline RP), inflammation, increased BALF concentrations of leukotriene (LT) C(4), LTD(4), and LTE(4) and prostaglandin (PG)D(2), and AHR. Pretreatment with aerosolized heparin attenuated late-phase airway obstruction by approximately 50%, inhibited eosinophil infiltration, reduced BALF concentrations of LTC(4), LTD(4), and LTE(4) and PGD(2), and abolished AHR. We conclude that heparin inhibits hyperventilation-induced late-phase changes in peripheral airway function, and does so in part via the inhibition of eosinophil migration and eicosanoid mediator production and release.

Inhibition of mast cell tryptase by inhaled APC 366 attenuates allergen-induced late-phase airway obstruction in asthma

Background: APC 366, a selective inhibitor of mast cell tryptase, has been shown to inhibit antigen-induced early asthmatic response (EAR), late asthmatic response (LAR), and bronchial hyperresponsiveness (BHR) in a sheep model of allergic asthma. Objective: The purpose of this study was to investigate the effects of APC 366 on antigen-induced EAR, LAR, and BHR in mild atopic asthmatics not on any anti-inflammatory therapy. Methods: Sixteen mild atopic asthmatics, each with a demonstrable antigen-induced EAR, LAR, and BHR to histamine, were recruited into this randomized, double-blinded, crossover study. APC 366 (5 mg)/placebo was administered by aerosol inhalation 3 times per day on treatment days 1 through 4. Allergen challenge was carried out on day 4. Histamine challenge was performed the following morning, 1 hour after final dosing. Results: Subjects were shown to have a significantly smaller overall mean area under the curve for the LAR (P = .012) and mean maximum fall in FEV1 for the LAR (P = .007) after pretreatment with APC 366 in comparison with placebo. No significant effects on BHR were demonstrable. Although the EAR was reduced by 18% after treatment with APC 366 in comparison with placebo, this was not statistically significant. Conclusion: Short-term repeated administration of APC 366 significantly reduced the magnitude of antigen-induced LAR in atopic asthmatics, which supports the role of mast cell tryptase in the pathophysiology of the LAR. (J Allergy Clin Immunol 2001;107:1039-45.)

Hypertonic saline aerosol increases airway reactivity in the canine lung periphery.

Hyperventilation with dry air increases airway surface fluid (ASF) osmolality and causes acute mucosal injury, leukocyte infiltration, and delayed airway obstruction and hyperreactivity in canine peripheral airways. The purpose of this study was to determine whether ASF hypertonicity per se can account for these hyperventilation-associated effects. We first measured ASF osmolality before and after normal (NSC) and hypertonic (HSC) saline aerosol challenges to document the magnitude of hypertonicity produced by these stimuli. We then measured canine peripheral airway resistance and reactivity to hypocapnia and aerosolized histamine before and after NSC and HSC. Cells and eicosanoid mediators recovered in bronchoalveolar lavage fluid at 5 and 24 h after NSC and HSC were examined. We found that HSC but not NSC caused acute ASF hyperosmolality, increased mediator release, and delayed airway hyperreactivity in the absence of mucosal injury and leukocyte infiltration. These observations suggest that ASF hyperosmolality contributes to the development of the late-phase response to hyperventilation and further suggest that hyperventilation-induced mucosal injury independently initiates leukocyte infiltration and late-phase airway obstruction.

Rapid Reversibility of the Allergen-Induced Pulmonary Late-PhaseReaction by an Intravenous β2-Agonist

Peebles R Jr, Permutt S, Togias AJ Appl Physiol.84199815001505To determine the degree to which β2-adrenergic receptor agonists can reverse the allergen-induced late reduction in lung function.Seven allergic subjects with mild asthma.In phase I of the study, terbutaline was administered as an initial bolus followed by continuous intravenous infusion over 45 minutes to determine the maximal attainable bronchodilation at baseline. In phase II, the subjects underwent an allergen inhalation (ragweed) challenge until forced expiratory volume in 1 second (FEV1) declined by at least 20%. Seven hours later, subjects were randomized and crossed over to receive terbutaline or a vehicle intravenous infusion identical to the phase I infusion protocol. Forced expiratory spirometry was performed at baseline and every hour after allergen-inhalation challenge. Spirometry was performed before and every 5 minutes through terbutaline (phase II and I) and placebo infusion (phase II) as well.At baseline, there was no significant difference in FEV1 or forced vital capacity (FVC) between phase I and phase II. In phase I, terbutaline infusion led to significant improvement in FEV1compared with preinfusion value (P < .03). After ragweed bronchoprovocation FEV1 decreased <85% at any single timepoint between hour 4 and 7 postallergen inhalation. On the day placebo was infused 7 hours after allergen inhalation, FEV1 values did not change significantly compared with the preinfusion value (P > .2). In contrast, on the day terbutaline was infused, FEV1 was significantly higher than preinfusion (P < .02). The overall pattern of FVC was the same as for FEV1. The kinetic curves for bronchodilator response to terbutaline infusion in phase I and in phase II were superimposable, indicating that the terbutaline induced bronchodilation was driven by the same pharmacologic mechanism.The late reduction in lung function caused by allergen inhalation challenge in asthmatic subjects is rapidly and almost completely reversible by an intravenous β2-adrenoreceptor agonist.Most previous studies have demonstrated that inhaled β2-adrenoreceptor agonists are only partially effective in reversing allergen-inhaled late phase airway obstruction. Interestingly, the ability to reverse this type of airway obstruction appears to be significantly enhanced using an intravenous route for drug administration. These observations challenge the current dogma that the airway obstruction observed 4 to 8 hours after allergen challenge is mostly related to airway edema, inflammation, and mucus secretion as opposed to bronchial smooth muscle spasm. A third arm of the study in which equivalent doses of β-agonist were given by aerosol would have been helpful in determining what role drug delivery may have contributed to the observed results.

Combined antagonism of leukotrienes and histamine produces predominant inhibition of allergen-induced early and late phase airway obstruction in asthmatics.

We defined the contribution of histamine and leukotrienes to allergen-induced airway obstruction in asthmatics; 12 subjects with allergic asthma underwent identical allergen bronchoprovocations on four occasions. At a control session, all subjects displayed early (EAR) and late asthmatic (LAR) reactions. The mean (+/- SE) drop in FEV1 during EAR (0-2 h) and LAR (2-12 h) was 29 +/- 2% and 28 +/- 4%, respectively. Thereafter, the influence of 1 wk randomized pretreatment with the leukotriene receptor antagonist zafirlukast (Accolate) (80 mg twice daily), the antihistamine loratadine (10 mg twice daily), and the combination of both antagonists was assessed. Expressed as AUC FEV1 in percent of the control reaction, zafirlukast reduced the response during EAR and LAR by 62 +/- 11% and 55 +/- 12%, respectively (p < 0.05 versus control). Loratadine inhibited EAR and LAR by 25 +/- 14% and 40 +/- 16%, respectively (p < 0.05 versus control). Zafirlukast was significantly more effective than loratadine during EAR but not during LAR. The combination of zafirlukast and loratadine reduced the AUC FEV1 during EAR and LAR further, by 75 +/- 8% and 74 +/- 14%, respectively (p < 0.05 versus control). The combination was significantly (p < 0.05) more effective than either drug alone during the LAR. The findings indicate that leukotrienes and histamine together mediate the major part of both the EAR and the LAR following exposure of asthmatics to allergen. Combination of leukotriene antagonism and antihistamines may represent a new strategy for treatment of airway obstruction in asthma.

Experimental Antiasthmatic Activity of RWJ 22108: A Bronchoselective Calcium Entry Blocker

RWJ 22108 (N-benzyl-N-methylaminoethyl 9-(2-chloro-6-fluorophenyl)-2,3,4,5,6,9-hexahydro-7-methyl-1, 1-dioxothiacyclohepteno-[3,2-b]pyridine-8-carboxylate) is a new bronchoselective calcium entry blocker with potential use as an antiasthmatic agent. Previous studies have shown that RWJ 22108 is a potent calcium entry blocker in vitro and demonstrates tissue selectivity for airway smooth muscle over vascular smooth muscle. The current study demonstrates the in vivo activity of RWJ 22108 in several different models of airway obstruction and asthma. RWJ 22108 relaxes preconstricted airways in dogs with little effect on blood pressure when administered by aerosol. In addition, it inhibits airway obstruction induced by antigen, histamine and exogenous leukotriene D4 in guinea pigs. In a conscious sheep model of allergic asthma, aerosol RWJ 22108 inhibits antigen-induced early and late phase airway obstruction and also the cellular infiltration associated with late phase. Total leukotrienes production is decreased in the guinea pig model probably as a result of fewer inflammatory cells infiltrating the lungs as shown in the sheep model of late phase. These data suggest that RWJ 22108 may have pharmacological potential in the clinical management of asthma.

Antigen-induced late-phase airway obstruction in the guinea pig?

Hartley or Dunkin-Hartley guinea pigs (n = 136) were actively sensitized to ovalbumin or Ascaris suum protein by five different regimes. Specific airway resistance (sR(AW)) was measured in conscious animals by a plethysmographic procedure before, immediately following and at various intervals (up to 96 h) after aerosolized antigen or vehicle challenge. Each sensitization and challenge regime produced an immediate allergic response with positive responses (defined as a 2-fold increase in sR(AW)) in 78-100% of animals. Recovery from the immediate response followed by late-phase responses was observed in only two out of a group of four animals. The results failed to substantiate literature reports of a high incidence of late responses in the guinea pig at 4-8, 17-24 or 72 h after allergen challenge.

A PAF receptor antagonist inhibits acute airway inflammation and late‐phase responses but not chronic airway inflammation and hyperresponsiveness in a primate model of asthma

We have examined the effects of a PAF receptor antagonist, WEB 2170, on several indices of acute and chronic airway inflammation and associated changes in lung function in a primate model of allergic asthma. A single oral administration WEB 2170 provided dose related inhibition of the release of leukotriene C4 (LTC4) and prostaglandin D2 (PGD2) recovered and quantified in bronchoalveolar lavage (BAL) fluid obtained during the acute phase response to inhaled antigen. In addition, oral WEB 2170 treatment in dual responder primates blocked the acute influx of neutrophils into the airways as well as the associated late-phase airway obstruction occurring 6 h after antigen inhalation. In contrast, a multiple dosing regime with WEB 2170 (once a day for 7 consecutive days) failed to reduce the chronic airway inflammation (eosinophilic) and associated airway hyperresponsiveness to inhaled methacholine that is characteristic of dual responder monkeys. Thus, we conclude that the generation of PAF following antigen inhalation contributes to the development of lipid mediators, acute airway inflammation and associated late-phase airway obstruction in dual responder primates; however, PAF does not play a significant role in the maintenance of chronic airway inflammation and associated airway hyperresponsiveness in this primate model.

Endothelial leukocyte adhesion molecule-1 mediates antigen-induced acute airway inflammation and late-phase airway obstruction in monkeys.

This study examines the role of endothelial leukocyte adhesion molecule-1 (ELAM-1) in the development of the acute airway inflammation (cell influx) and late-phase airway obstruction in a primate model of extrinsic asthma. In animals sensitive to antigen, a single inhalation exposure induced the rapid expression of ELAM-1 (6 h) exclusively on vascular endothelium that correlated with the influx of neutrophils into the lungs and the onset of late-phase airway obstruction. In contrast, basal levels of ICAM-1 was constitutively expressed on vascular endothelium and airway epithelium before antigen challenge. After the single antigen exposure, changes in ICAM-1 expression did not correlate with neutrophil influx or the change in airway caliber. This was confirmed by showing that pretreatment with a monoclonal antibody to ICAM-1 did not inhibit the acute influx of neutrophils associated with late-phase airway obstruction, whereas a monoclonal antibody to ELAM-1 blocked both the influx of neutrophils and the late-phase airway obstruction. This study demonstrates a functional role for ELAM-1 in the development of acute airway inflammation in vivo. We conclude that, in primates, the late-phase response is the result of an ELAM-1 dependent influx of neutrophils. Therefore, the regulation of ELAM-1 expression may provide a novel approach to controlling the acute inflammatory response, and thereby, affecting airway function associated with inflammatory disorders, including asthma.

Cyclosporine A inhibits allergen-induced late asthmatic response and increase of airway hyperresponsiveness in guinea pigs

Considerable interest has recently focused on the role of T-cell function in the pathogenesis of asthma. We have previously demonstrated that repeated inhaled antigen (ovalbumin; OA) exposure resulted in an appearance of late phase airway obstruction (LAR) in more than 50% and significant increase of airway hyperresponsiveness (AH) in guinea pig experimental models. We have studied the effect of cyclosporine A (CsA), a potent helper T-cell suppressant, on these models in vivo. Respiratory resistance (Rrs) of sensitized guinea pigs by repeated OA inhalation was measured by the oscillation method and AH estimated as an inhaled concentration of histamine, causing a 200% increase in the baseline Rrs (PC200 Hist). The magnitude of immediate OA (10 mg/ml/min) inhalation-induced bronchoconstriction was not significantly different in CsA-treated (25 mg/kg/day, 7-day oral administration) and non-treated groups. However, the development of LAR was markedly inhibited in CsA treatment groups (n = 5). Antigen-induced increase of AH at 24 hr and 5 days was also significantly inhibited by CsA pretreatment. We conclude that CsA is capable of inhibiting the development of LAR and increase of AH, and thus the regulation of T-cell function may contribute to the treatment of asthma.

Dry air-induced late phase responses in the canine lung periphery

Although controversial, late phase responses in asthmatic subjects have been reported several hours after exercise. We previously showed that exposure to dry air increases collateral system resistance (Rcs) in the canine lung periphery, and produces acute airway responses analogous to those that characterize human exercise-induced asthma. We used a dual wedged bronchoscope technique in anaesthetized male mongrel dogs to monitor Rcs in: 1) control segments continuously exposed to 200 ml.min-1 of 5% CO2 in air and 2) dry air challenged segments exposed to 2000 ml.min-1 5% CO2 for 5 min. We examined Rcs at 5 min and approximately 5 h post-challenge in an attempt to document late phase airway obstruction. Five min after dry air challenge Rcs initially increased 114 +/- SE 22%; contralateral control segments remained unchanged (n = 9). Five hour post-challenge, Rcs in dry air exposed segments was elevated 81 +/- 20% above pre-challenge baseline (p less than 0.01); contralateral control segments did not change significantly over the 5 h period. Cell profile analyses of lavage samples at 5 hours revealed that neutrophils and eosinophils were significantly increased (p less than 0.03) in dry air challenged segments when compared to controls. Leukotriene C4/D4 concentration in lavage was correlated (p less than 0.02) with neutrophil infiltration. Thus, we conclude that the canine lung periphery represents a reproducible model of a dry air-induced late phase reaction.