Elsevier

Cellular Immunology

Volume 219, Issue 2, October 2002, Pages 92-97
Cellular Immunology

Attenuation of airway hyperresponsiveness in a murine asthma model by neutralization of granulocyte–macrophage colony-stimulating factor (GM-CSF)

https://doi.org/10.1016/S0008-8749(02)00565-8Get rights and content

Abstract

Asthma is recognized as an inflammatory disease in which various cytokines are involved. Among these, granulocyte–macrophage colony-stimulating factor (GM-CSF) is known to play a critical role in the survival of eosinophils and in the activation of antigen-presenting cells (APC). We studied the effects of neutralization of GM-CSF in a murine model of asthma, to elucidate its role in enhanced airway responsiveness and in airway inflammation. A/J mice, which are genetically predisposed to acetylcholine hyperresponsiveness, were immunized with ovalbumin (OA) and alum. Thereafter, the mice were subjected to a two-week regimen of OA inhalation, during which either goat anti-mouse polyclonal GM-CSF antibody or isotype control goat IgG was administered intranasally. Pulmonary function was then analyzed using whole body plethysmography before and after acetylcholine (Ach) inhalation. Here we show that OA inhalation following OA immunization increased airway responsiveness to acetylcholine and induced GM-CSF as well as IL-4 and IL-5 mRNA expression in the lung. The administration of GM-CSF-neutralizing antibody during OA inhalation significantly reduced this increased airway hyperresponsiveness and also inhibited airway inflammation. Thus, endogenous GM-CSF plays an important role in the process of airway inflammation and airway hyperresponsiveness after antigen-specific immunity has been established.

Introduction

Various cytokines, especially the Th2 cytokines IL-4, IL-13, and IL-5, have been reported to play causative roles in allergic inflammation and airway hyperresponsiveness in asthma [1], [2], [3], [4], [5]. Granulocyte–macrophage colony-stimulating factor (GM-CSF), which is produced by both Th2 and Th1 cells, is also a key cytokine for eosinophil survival, as is IL-5. GM-CSF is responsible for promoting the differentiation of eosinophils from promyelocytes [6], [7]. It also activates eosinophils and prolongs eosinophil survival in the peripheral tissues [8], [9]. In addition, GM-CSF is also a key cytokine for growth and differentiation of antigen-presenting cells (APC) [10], [11]. It has been shown that hyperexpression of GM-CSF promotes sensitization to allergens, resulting in augmentation of airway inflammation [12], [13]. Because GM-CSF is produced by macrophages, eosinophils and epithelial cells at the site of asthma [14], [15], endogenous production of GM-CSF may also play an important role in the pathogenesis of asthma.

We previously report that GM-CSF plays a central role in airway hyperresponsiveness caused by diesel exhaust particulates (DEP), a significant air pollutant [16]. Intranasal administration of antibody raised against GM-CSF, but not against IL-4, abolished DEP-evoked increases in airway responsiveness and mucus-secreting cell hyperplasia. In the present study, we aimed to clarify the effects of endogenous GM-CSF in airway hyperresponsiveness induced in an allergen-dependent manner. We found that both airway hyperresponsiveness and mucus-producing cell hyperplasia are significantly inhibited by a GM-CSF-specific neutralizing antibody administered during aeroallergen exposure. Our study indicates that GM-CSF plays a critical role in the development of airway hyperresponsiveness and therefore may represent a therapeutic target.

Section snippets

Reagents

Goat anti-mouse GM-CSF-neutralizing antibodies were purchased from R&D Systems (Minneapolis, MN); This antibody (0.1μg/ml) neutralizes 0.8 ng/ml mGM-CSF. The antibody dose was 50μg/mouse. Goat IgG was from Chemicon International (Temecula, CA), pentobarbital sodium from Pitman More (Mundelein, IL), and pancuronium bromide from Organ (Teknika, The Netherlands).

Sensitization

Male A/J mice (5 weeks old), which have native airway hyperresponsiveness to acetylcholine [17], [18], were purchased from SLC (Sizuoka,

Bronchial hyperresponsiveness induced by OA inhalation

First, we determined whether OA inhalation did affect airway hyperresponsiveness in A/J mice, which carry a genetic background conferring hypersensitivity to acetylcholine [17], [18]. As shown in Fig. 1, there is a significant increase in AHR in the OA-immunized/non-challenged mice (▴) compared to the non-immunized control mice (•) after inhalation of 5.0 mg/ml Ach (p<0.05). There was, however, a striking increase in AHR in the OA inhalation group (□) compared with the other two groups after

Discussion

In the study presented here, we have shown in a mouse model of asthma that allergic inflammation and airway responsiveness to acetylcholine were abolished by neutralization of endogenous GM-CSF during aeroallergen exposure. It has been established that GM-CSF possesses adjuvant effects when administered during immunization, because it is critical for the maturation of dendritic cells [12], [13], [22], [23]. Because antigen presentation is not only required during immunization but also for

Acknowledgments

We wish to thank Miss Yukiko Matsuo and Miss Maya Kuwabara for their expert technical assistance.

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