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ABSTRACT |
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ABSTRACT
INTRODUCTION
METHODS
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DISCUSSION
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In the guinea pig, interleukin-5 (IL-5) has been shown to induce airway hyperresponsiveness as well as eosinophilia, which are important symptoms in asthma. IL-5 seems to be a critical cytokine since it selectively affects eosinophil functions. The mechanism of action by which IL-5 leads to airway hyperresponsiveness may be important for our understanding of the pathogenesis of asthma. Neurogenic inflammation, which is mediated by nonadrenergic noncholinergic nerves (NANC), may play a role in the IL-5-induced effects in guinea pig airways. In this study, the role of neuropeptides in the IL-5- induced airway hyperresponsiveness and eosinophilia in the guinea pig was examined using selective neurokinin receptor antagonists. Intra-airway application of IL-5 (1 µg, twice) induces a selective eosinophil migration (control: 12 [8 -22] × 105 cells and IL-5: 90 [67-187] × 105 cells, p < 0.05) and activation (control: 6.3 ± 0.9 ng eosinophil peroxidase [EPO]/ml bronchoalveolar lavage [BAL] fluid and IL-5: 29.3 ± 4.9 ng EPO/ml BAL fluid, p < 0.05) and a pronounced airway hyperresponsiveness in vivo. The maximal responses to histamine are increased by 160 ± 16% (p < 0.05) after IL-5. Treatment of guinea pigs with either the nonselective neurokinin (NK)-receptor antagonist, FK224, or the selective NK2-receptor antagonist, SR48968, results in a complete inhibition of the in vivo hyperresponsiveness found after application of IL-5. Vice versa, intra-airway administration of substance P (10 µg, twice) results in an airway hyperresponsiveness (increased maximal response after substance P: 166 ± 15% [p < 0.05]) without inducing migration or activation of eosinophils. All examined NK- receptor antagonists do not influence the IL-5-induced eosinophil accumulation. In addition, no effect of the NK-receptor antagonists is observed on the IL-5-induced eosinophil activation, as determined by BAL fluid EPO levels. The release of NK2-receptor active tachykinins plays an important role in the development of IL-5-induced airway hyperresponsiveness. This feature appears to be a step following eosinophil infiltration and activation since there are no effects on eosinophil function by pretreatment of the used NK-receptor antagonists.
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INTRODUCTION |
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ABSTRACT
INTRODUCTION
METHODS
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Infiltration into the airways of inflammatory cells, particularly T lymphocytes and eosinophilic granulocytes, and pulmonary hyperresponsiveness to bronchoconstrictor mediators are prominent features of the pathogenesis of asthma (1). However, the exact mechanistic relationship between airway hyperresponsiveness and leukocyte infiltration is still unknown. Recent studies suggest a role for T helper 2 lymphocyte-derived cytokines such as interleukin-5 (IL-5) in the development of airway eosinophilia and hyperresponsiveness (2, 3). We and others have demonstrated that antibodies to IL-5 inhibited the airway hyperreactivity and bronchoalveolar eosinophilia in a guinea pig model for allergic asthma (4, 5). IL-5 is reported to be a selective activator of eosinophils, which promotes growth, differentiation, proliferation and survival of eosinophils (6, 7). In addition, IL-5 is an eosinophilic chemoattractant and primes eosinophils for enhanced chemotaxis and leukotriene production in response to several inflammatory mediators (8). Therefore, this cytokine may be responsible for the selective recruitment and activation of eosinophils at sites of allergic reactions.
Indeed, administration of IL-5 has been demonstrated to induce airway eosinophilia and hyperreactivity in guinea pigs and mice (4, 11). Very recently, in the guinea pig we have shown that for the development of IL-5-induced airway hyperresponsiveness in vivo, the very late activation antigen-4 (VLA-4) dependent infiltration of eosinophils into the bronchial tissue is essential (14). VLA-4 is an important and selective adhesion molecule in the infiltration of eosinophils, but not neutrophils, from the vasculature to the airway tissue. These data underline the putative role of eosinophils in the development of pulmonary hyperresponsiveness.
In the airways it has been well recognized that nonadrenergic noncholinergic nerves (NANC) form part of the local nervous system. Neurogenic inflammation, which is mediated by NANC nerves, may play a role in asthma (15). It is now known that NANC neuropeptides, such as the tachykinins, substance P, or neurokinin A, have potent inflammatory effects and can affect airway function in a way which resembles features found in the pathogenesis of asthma. Several studies have reported that exposure of guinea pigs to an aerosol of either capsaicin, a substance releasing endogenous NANC neuropeptides, or substance P elicited airway hyperresponsiveness to bronchoconstrictor agents (16). In addition, in a guinea pig model for allergic asthma, depletion of NANC neuropeptides by chronic capsaicin pretreatment, resulted in a profound inhibition of antigen-induced airway hyperresponsiveness (19, 20). However, the antigen-induced eosinophilia was still evident (20). In addition, van Oosterhout and colleagues have shown in the guinea pig that after in vivo administration of either IL-5 or substance P, tracheal responses to histamine were significantly increased (13). Application of IL-5, but not of substance P, induced a significant increase in the number of eosinophils as well as a rise in eosinophil peroxidase activity (a marker for eosinophil activation) in bronchoalveolar lavage fluid of guinea pigs (13). These results suggest that IL-5 is important in the recruitment and activation of eosinophils, whereas NANC neuropeptides seem to be involved in the process by which eosinophils induce hyperreactivity.
To investigate the role of tachykinins in the development of changes in airway function, the effect of several neurokinin receptor antagonists was studied on IL-5-induced airway hyperresponsiveness and eosinophilia in the guinea pig.
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ABSTRACT
INTRODUCTION
METHODS
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DISCUSSION
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Animals
The animals used in this study were specified pathogen-free male Dunkin Hartley guinea pigs weighing 390 -550 g (Harlan Porcellus, Blackthorn, Bicester, Oxon, UK). Water and commercial chow were allowed ad libitum. The guinea pigs were free of respiratory infections as assessed by the health monitoring quality control report by Harlan Porcellus. The experiments were approved by the Animal Care Committee of the Utrecht University (Utrecht, The Netherlands).
Intra-airway Administration of IL-5 or Substance P
Guinea pigs received either 1 µg IL-5, 10 µg substance P, or vehicle consisting of saline with 0.1% bovine serum albumin (BSA) in a volume of 300 µl intranasally under short-lasting anesthesia (Ketalar®, 40 mg/kg intramuscularly, Parke-Davis, Hoofddorp, The Netherlands; and Rompun®, 5 mg/kg subcutaneously, Bayer BV, Mijdrecht, The Netherlands) twice on one day (9:00 A.M. and 4:00 P.M.).
Neurokinin Receptor Antagonist Treatment
The nonselective neurokinin receptor antagonist, FK224, and the neurokinin-1 receptor antagonist, FK888, were dissolved in 0.1% ethanol and injected intravascularly at a dose of 1 µmol/kg and 2 µmol/kg body weight, respectively, 30 min before each IL-5 or vehicle administration. The selective neurokinin-2 receptor antagonist, SR 48698, was dissolved in saline and injected intraperitoneally at a dose of 1 mg/kg body weight, 1 h before each IL-5 or vehicle administration. The dose and route of administration of neurokinin receptor antagonists were selected based on results showing the effects of FK224, FK888, or SR 48968 on tachykinin- or antigen-induced bronchoconstriction in guinea pigs (21). In addition, in a pilot experiment it was demonstrated that pretreatment with FK888 (1 µmol/kg body weight) resulted in a 61.9 ± 0.3% (mean ± STD, n = 2) of substance P-induced decrease in blood pressure.
Hirayama and colleagues (23) examined the non-specific actions of FK224 and FK888 on PAF-induced plasma exudation and bronchoconstriction in capsaicinized guinea pigs. They have demonstrated a complete lack of inhibitory effect of FK224 and FK888. Moreover, in the guinea pig, it is shown that SR48968 did not affect acetylcholine-induced bronchoconstriction or histamine-induced effects on airway resistance and blood pressure (21, 37). Thus it can be concluded that there are no nonspecific actions for the antagonists used in this study.
Airway Reactivity in vivo
Twenty-four hours after the first administration of IL-5 or vehicle, the
guinea pigs were prepared for measurement of lung resistance (RL).
The animals were anesthetized with urethane (2.8 g/kg intraperitoneally) and were able to breathe spontaneously. To avoid an anesthesia-induced fall in body temperature the animals were placed in a
heated chamber at approximately 30° C. To determine airflow (V)
and tidal volume (VT) the trachea was cannulated and connected to a
Gould Godart pneumotachograph (Gould Godart, Bunnik, The Netherlands) with a Fleisch flow head (No. 000; Meijnhart, Bunnik, The Netherlands). A Validyne MP45-24 pressure transducer (Validyne Engineering Corp., Northridge, CA) measured the pressure difference between
the tracheal cannula and a cannula filled with saline inserted in the
esophagus, which presented the transpulmonary pressure (TPP). RL
was determined breath by breath by the method of Amdur and Mead
(24) using a computerized respiratory analyzer. Dividing 
TPP by V
at isovolume points (50%) yielded the RL. A small polyethylene catheter used for intravenous administration of histamine was placed in
the right jugular vein. A histamine dose-response (2 to 20 µg/kg)
curve was made. Injections were given at intervals of at least 5 min in
which RL returned to baseline. Responses are presented as increases
in RL above baseline.
Bronchoalveolar Lavage
Bronchoalveolar lavages (BAL) were performed in all guinea pigs
used. After measurement of lung resistance, the animals received a lethal dose of pentobarbital sodium (300 mg/kg intraperitoneally). Via
the tracheal cannula, the lungs were filled in situ with 5 to 10 ml NaCl-EDTA-buffer (0.15 M NaCl, 2.6 mM EDTA) using a syringe. Fluid
was withdrawn from the lungs and collected in a plastic tube on ice.
The first lavage (approximately 5 ml) recovered from each animal was
kept separate, and the lavages thereafter were pooled until 50 ml fluid
was obtained. The cells were sedimented by centrifugation at 400 g for
10 min at 4° C. The supernatant of the first lavage (cell-free BAL
fluid) was stored at 
70° C prior to analysis of BAL fluid eosinophil
peroxidase activity. The two cell pellets from the lavages were resuspended and pooled. The cells were washed twice with NaCl-EDTA-buffer. A sample of the cells were stained with Turk solution and
counted. All cell preparations were analyzed morphologically after
centrifugation on microscopic slides. Air-dried preparations were
fixed and stained with Diff-Quik (Merz & Dade A.G., Dudingen,
Switzerland). Differential counts were made under oil immersion microscopy.
BAL Fluid Eosinophil Peroxidase Activity
The eosinophil peroxidase (EPO) activity in the supernatant of the first lavage (cell-free BAL fluid) was measured according to the method of Strath and coworkers (25), which is based on the oxidation of O-phenylenediamine (OPD) by EPO in the presence of hydrogen peroxide (H2O2). The substrate solution consisted of 10 mM OPD in 0.05 M Tris-buffer (pH = 8) and 4 mM H2O2 (BDH, Poole, UK). Substrate solution (100 µl) was added to BAL cell supernatant samples (50 µl) in a 96-wells microplate and incubated at room temperature for 30 min before stopping the reaction by addition of 50 µl of 4 M sulfuric acid. The absorbance was then measured at 492 nm using a Titerek Multiskan (Flow Labs., Irvine, UK). Duplicate incubations were carried out in the absence and presence of the EPO inhibitor 3-amino-1,2,4-triazole (AMT, 2 mmol/l). Blanks were cell free BAL fluid samples (50 µl) incubated with Tris-HCl buffer. Serial dilutions of horseradish peroxidase (200 ng/ml) were used to quantitate the amount of peroxidase in the samples. Results are expressed as ng/ml peroxidase activity and were corrected for the activity of other peroxidases, which were not inhibitable by AMT.
Materials
Substance P, O-phenylenediamine, 3-amino-1,2,4-triazole, horseradish peroxidase, and bovine serum albumin grade V were obtained from Sigma Chemical Company (St. Louis, MO). The human recombinant IL-5 was purified by Dr. A. E. Proudfoot (Glaxo Institute for Molecular Biology, Geneva, [26]) and kindly donated by Dr. D. Fattah (Glaxo, Greenford, UK). The dipeptide neurokinin-1 receptor antagonist FK888, N2[(4R)-4-hydroxy-1-(1-methyl-1H-indol-3-yl)carbonyl-L-propyl]-N-methyl-N-phenylmethyl-3-(2-naphtyl)L-alaninamide, and the peptidergic neurokinin1/2 receptor antagonist, FK224, N-(N2-(N-(N-(N-(2,3-didehydro-N-methyl-N-(N-(3-(2-penthylphenyl)- propionyl)-L-threonyl)tyrosyl-L-leucynyl)-D-phenylalanyl)-L-allo-threonyl)-L-asparaginyl-L-serine-v-lactone, were gifts from Fujisawa Pharmaceuticals Co. Ltd. (Osaka, Japan). SR 48968, (S)-N-methyl-N(4-acetylamino-4-phenylpiperidino-2-(3,4-dichloophenyl)butyl)benzamide, was kindly donated by Sanofi Recherche (Montpellier, France).
Data Analysis
The cellular accumulation in BAL fluid was analyzed by using a distribution-free Kruskal-Wallis one-way analysis of variance test. The cell data are expressed as medians (minimum-maximum).
The EPO activity data and airway reactivity data are expressed as mean ± SEM. These data were initially analyzed using Bartlett's test for homogeneity of variances. For the raw EPO activity data the Bartlett's test indicated no heterogeneity of variance. Analysis of the raw airway resistance data with the Bartlett's test demonstrated a significant difference between group variances. However, the group variances of log-transformed data were homogenous. All subsequent analyses were done on raw EPO activity data and log-transformed airway resistance data using the parametric analysis of variance (ANOVA). Differences between groups were tested with a Bonferonni post-hoc test. A p value < 0.05 was considered to reflect a statistically significant difference. All analyses were performed by using SYSTAT (version 5.03, Wilkinson. Leland. SYSTAT: the system for statistics. Evanston, IL., Systat Inc., 1990).
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RESULTS |
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INTRODUCTION
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IL-5-induced Changes in Guinea Pig Airways
One day after intra-airway application of IL-5 (twice: 1 µg) bronchial hyperresponsiveness to histamine in vivo in the guinea pig was observed (Figure 1). In IL-5-treated animals a dose of 20 µg/kg histamine resulted in an increase in lung resistance of 160 ± 16% when compared with vehicle treatment (p < 0.05, n = 5, ANOVA). The change in bronchial reactivity was associated with a marked accumulation of eosinophils into the airways (Table 1). Furthermore, an enhanced EPO activity in cell free BAL fluid (control: 6.3 ± 0.9 ng/ml and IL-5: 29.3 ± 4.9 ng/ml, p < 0.05, n = 5, ANOVA) was observed after intra-airway administration of IL-5 indicating that the eosinophils were activated.
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Substance P-induced Changes in Guinea Pig Airways
One day after intra-airway administration of substance P (twice: 10 µg) the increases in airway resistance to increasing doses of histamine were significantly enhanced (Figure 1). The maximal increase in lung resistance to a dose of 20 µg/kg histamine potentiated with 166 ± 15% in substance P-treated animals when compared with vehicle-treated animals (p < 0.05, n = 5, ANOVA). The substance P-induced airway hyperresponsiveness was not accompanied by a rise in number of eosinophils in the BAL fluid nor by changes in EPO activity in cell free BAL fluid (Table 1 and Figure 2).
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Effects of Neurokinin Receptor Antagonists on IL-5-induced Changes in Guinea Pig Airways
The effects of the non-selective neurokinin (NK) receptor antagonist, FK224, the neurokinin-1 (NK1) receptor antagonist, FK888, and the neurokinin-2 (NK2) receptor antagonist, SR48968, were studied on the IL-5-induced bronchial hyperresponsiveness, eosinophil accumulation and activation in the guinea pig airways. Pretreatment of the guinea pigs with either FK224 (NK receptor antagonist) or SR 48968 (NK2 receptor antagonist) resulted in a complete inhibition of the in vivo bronchial hyperresponsiveness found 24 h after intra-airway administration of IL-5 (Figures 3B and 4). In contrast, the NK1 receptor antagonist, FK888, did not influence the IL-5-induced responses on airway resistance to histamine in the guinea pig (Figure 3C). The three examined NK receptor antagonists did not influence airway resistance to histamine in control animals. There were no statistically significant differences between all control groups.
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All examined NK receptor antagonists did not affect the IL-5-induced eosinophil accumulation into the BAL fluid (Figure 5). After pretreatment with FK224, FK888, or SR 48968, intra-airway administration of IL-5 resulted in a comparable and significant increase in the numbers of eosinophils in the BAL fluid when compared with vehicle pretreatment. In addition, no effects of the NK receptor antagonists were observed on the IL-5-induced increase in EPO activity in the cell free BAL fluid of the guinea pig airways (Figure 6).
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DISCUSSION |
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In this study, we have demonstrated that the neurokinin-2 receptor plays an important role in the development of IL-5- induced hyperresponsiveness but not in the eosinophil infiltration into the airways of guinea pigs.
The T helper 2 lymphocyte-derived cytokine IL-5 has been shown to induce bronchial hyperresponsiveness as well as airway eosinophilia, which are important symptoms in asthma (4, 11, 27). In asthmatic patients, IL-5 is produced locally in the airways and this phenomenon is related to the number of eosinophils in the airways (3, 28). Furthermore, in allergic asthma mRNA expression of T helper 2 cytokines, in particular IL-5, is associated with clinical measures of asthma severity, i.e., airflow restriction and hyperresponsiveness to bronchoconstrictor agents (28, 29). Although IL-5 probably acts in concert with other cytokines, IL-5 emerges as a critical cytokine in asthma. To understand the importance of IL-5 in the pathogenesis of asthma, it is very relevant to try to elucidate the possible mechanism of action of IL-5.
In the guinea pig, we have demonstrated that IL-5 induces selective migration and activation of eosinophils and a pronounced airway hyperresponsiveness to histamine in vivo. Similar results were also obtained by other investigators (11, 13, 30). This relatively simple model of IL-5-induced airway hyperresponsiveness enables us to investigate the correlation between eosinophil infiltration and the development of bronchial hyperresponsiveness. This is in contrast to animal models for asthma such as ovalbumin-sensitized and -challenged guinea pigs, where besides the eosinophil a whole range of inflammatory cells are activated (4).
In the airways, it has been well recognized that NANC nerves form part of the local nervous system. Neurogenic inflammation, which is mediated by NANC nerves, may play a role in asthma (15). It is now known that NANC neuropeptides, such as the tachykinins, substance P or neurokinin A, have potent inflammatory effects and can affect airway function in a way that resembles features found in the pathogenesis of asthma. In several animal species, the release of endogenous neuropeptides, elicited by inhalation of capsaicin aerosols, as well as inhaled or infused exogenous tachykinins are able to induce bronchoconstriction and airway hyperresponsiveness to contractile agents (16 -18). Immunohistochemical studies of neuronal substance P in airways of asthmatic subjects have yielded conflicting results. While in some studies in asthmatic patients, an increase in both number and length of tachykinin-immunoreactive nerve fibers was found in the airways when compared with nonasthmatic subjects (31, 32), others detected significantly less substance P-like immunoreactivity in lung tissue from asthmatic than from nonasthmatic patients (33, 34). However, this latter finding may reflect augmented substance P release. Moreover, in asthma patients substance P enhances maximal airway narrowing to methacholine in patients 24 h after inhalation (35).
In the guinea pig, several investigators have demonstrated that depletion of NANC neuropeptides by chronic capsaicin pretreatment prevents the development of allergen-induced hyperresponsiveness despite the presence of eosinophils (19, 20), while others have not (36, 37). In addition, Fischer and coworkers have shown that sensitization of guinea pigs to ovalbumin induced a 1 to 5-fold increase of neuropeptide concentration in lung tissue and also increased 2-fold tachykinin-immunoreactive neurons 24 h after the challenge (38). Van Oosterhout and colleagues reported that in the guinea pig after in vivo administration of either IL-5 or substance P, tracheal responses to histamine were significantly increased in vitro (13). These results are in accord with our findings with IL-5 or substance P on in vivo airway function. In contrast to the IL-5-induced hyperresponsiveness, the substance P-induced hyperresponsiveness was not accompanied by changes in the number of eosinophils in BAL fluid. In addition, substance P did not induce eosinophil degranulation since the EPO activity in cell free BAL fluid was not altered. Since the IL-5 and substance P induce airway hyperresponsiveness to a similar extent in vivo after intra-airway administration (this study) and since both responses in vitro were not additive (13), the mechanism of action of the two agents may be similar. All these data suggest that IL-5 is important in the recruitment and activation of eosinophils, whereas tachykinins seem to be involved more downstream in the sequence by which eosinophils induce hyperresponsiveness. In this study, the role of neuropeptides in the IL-5-induced airway hyperresponsiveness and eosinophilia was examined using selective neurokinin receptor antagonists. Three types of tachykinin receptors have been characterized namely NK1, NK2, and NK3 (15). These receptors are preferentially activated by substance P, neurokinin A, and neurokinin B, respectively. The tachykinin receptor antagonists studied were: NK1/NK2-receptor antagonist, FK224, and NK1-receptor antagonist, FK888, and NK2-receptor antagonist, SR48968. Our results demonstrate that in the guinea pig the NK2 receptor mediates the hyperresponsiveness found 24 h after intranasal IL-5 administration and that neither the NK1 nor the NK2 receptor is involved in the IL-5-induced eosinophil migration and activation. In addition, the tachykinin substance P was not able to induce eosinophilia in the guinea pig airways. Foulon and colleagues demonstrated that in the guinea pig, NKA and substance P produce bronchoconstriction through NK2 and NK1 receptor stimulation, respectively (39). A role for NK2 receptor in the development of bronchial hyperresponsiveness is further supported by studies in guinea pig and man showing that neurokinin A is far more potent than substance P in contracting airway smooth muscle (21, 40- 42). In addition, in lung samples containing membranous airways, NK2-receptor mRNA expression was increased fourfold in asthmatics compared with controls, whereas NK1 receptor mRNA levels were similar in the two groups (43), suggesting that the NK2 receptor could play an important role in the development of bronchial hyperresponsiveness.
Several explanations can be suggested to explain the link between the role of eosinophils and tachykinins in the development of IL-5-induced hyperresponsiveness in the guinea pig. First, IL-5 could induce bronchial hyperresponsiveness via a direct action on sensory nerve endings, completely independent of eosinophil migration/activation. Recently, however, we have demonstrated that for the development of IL-5-induced airway hyperresponsiveness in the guinea pig the VLA-4 dependent infiltration of eosinophils is essential (14).
Second, tachykinins may be involved in the activation of eosinophils that are already primed by IL-5. Indeed, Kroegel and colleagues have demonstrated that substance P can induce EPO release from isolated guinea pig eosinophils (44). This tachykinin-induced EPO release, however, was not mediated via a neurokinin receptor-dependent mechanism. For this reason, it is not surprising that in our study the selective neurokinin receptor antagonists did not have any effect on the eosinophil activation after IL-5 administration. In addition, in our study, intra-airway application of substance P did not result in eosinophil activation in vivo indicating a more down-stream role for tachykinins in the development of airway hyperresponsiveness.
Third, it could be possible that mediators from IL-5-activated eosinophils may induce the release of neuropeptides from sensory nerves in the airways. Subsequently, substance P or neurokinin A may induce airway hyperresponsiveness via the NK2 receptor on bronchial smooth muscle cells. Indeed, Garland and colleagues have demonstrated that mediator release from activated eosinophils can directly stimulate tachykinin release from sensory C-fiber neurons in cell culture (45). Released cationic granule proteins from eosinophils are likely mediators, since it has been shown that they can induce the release of neuropeptides in human bronchi, indicating the activation of NANC nerves (46). In addition, intratracheal instillation of cationic proteins induced airway hyperreactivity in rats (47). This change in airway function could be antagonized with a neurokinin receptor antagonists. Moreover, it has recently been shown that NK2-receptor antagonist, SR 48968, prevents ovalbumin-induced airway hyperreactivity in sensitized guinea pigs (21). In line with these data, it has been demonstrated that airway nerves are surrounded by and infiltrated with eosinophils after ovalbumin challenge (48). These studies together with our results are consistent with an effect of eosinophils on airway neural function.
In conclusion, IL-5 may be an inflammatory mediator primarily involved in the recruitment and activation of eosinophils and the tachykinins, substance P or neurokinin A, have a more downstream role in the sequence by which eosinophils induce airway hyperresponsiveness. Interestingly, in several animal models of asthma, such as ovalbumin-, ozone- and dinitrobenzene-induced hypersensitivity and respiratory viral infections, the development of airway hyperresponsiveness is also dependent on the release of sensory neuropeptides indicating that neuropeptides seem to be a common step in the pathway leading to airway hyperresponsiveness (20, 49). Selective neurokinin receptor antagonists or drugs that inhibit the release of neuropeptides could be expected to have a beneficial effect in the pathogenesis of asthma by reducing the neurogenic component of inflammation. However, the future therapeutic potential of such pharmacological agents remains to be investigated in the clinic.
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Footnotes |
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Correspondence and requests for reprints should be addressed to Aletta D. Kraneveld, Department of Pharmacology and Pathophysiology, Utrecht Institute of Pharmaceutical Sciences, Utrecht University, P.O. Box 80.082, 3508 TB Utrecht, The Netherlands. E-mail: A.D.Kraneveld{at}FAR.RUU.NL
(Received in original form August 26, 1996 and in revised form November 7, 1996).
Acknowledgments: This study was supported by a research grant (94.97) of the Dutch Asthma Foundation.
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References |
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