Effects of in vitro exposure to hay dust on expression of interleukin-17, -23, -8, and -1β and chemokine (C-X-C motif) ligand 2 by pulmonary mononuclear cells isolated from horses chronically affected with recurrent airway disease

Article · January 2008with56 Reads
DOI: 10.2460/ajvr.68.12.1361 · Source: PubMed
To examine effects of in vitro exposure to solutions of hay dust, lipopolysaccharide (LPS), or beta-glucan on cytokine expression in pulmonary mononuclear cells isolated from healthy horses and horses with recurrent airway obstruction (RAO). 8 RAO-affected and 7 control horses (experiment 1) and 6 of the RAO-affected and 5 of the control horses (experiment 2). Bronchoalveolar lavage cells were isolated from horses that had been stabled and fed dusty hay for 14 days. Pulmonary mononuclear cells were incubated for 24 (experiment 1) or 6 (experiment 2) hours with PBS solution or solutions of hay dust, beta-glucan, or LPS. Gene expression of interleukin (IL)-17, IL-23(p19 and p40 subunits), IL-8, IL-1beta, and chemokine (C-X-C motif) ligand 2 (CXCL2) was measured with a kinetic PCR assay. Treatment with the highest concentration of hay dust solution for 6 or 24 hours increased expression of IL-23(p19 and p40), IL-8, and IL-1beta in cells from both groups of horses and increased early expression of IL-17 and CXCL2 in RAO-affected horses. Lipopolysaccharide upregulated early expression of IL-23(p40) and IL-8 in cells from both groups of horses but only late expression of these cytokines in cells from RAO-affected horses. Treatment with beta-glucan failed to increase cytokine expression at 6 or 24 hours. Cells from RAO-affected horses were not more responsive to the ligands tested than were cells from control horses, which suggests a minimal role of mononuclear cells in propagation of airway neutrophilia in horses with chronic RAO.
    • similar role on neutrophils. We chose an equine model for our study due to the fact that horses are prone to developing a severe neutrophilic asthma-like condition associated with an upregulation of IL-17 within their airways [15, 25], in which the neutrophilic inflammation is poorly responsive to GCs [16, 26], as seen in some humans [27, 28]. In a response similar to that of human airway epithelial cells [12] , IL-17-induced an upregulation of IL-8 mRNA, a potent neutrophil chemokine [29], in neutrophils stimulated for 24 hours.
    [Show abstract] [Hide abstract] ABSTRACT: Glucocorticoids (GCs) are the most effective drugs for the treatment of human asthma. However, a subgroup of asthmatic patients with neutrophilic airway inflammation is insensitive to GCs. Interleukin-17 (IL-17), a cytokine upregulated in the airways of a subset of human asthmatic patients, contributes to the recruitment of neutrophils and induces a glucocorticoid resistance in human airway epithelial cells. We hypothesized that IL-17 similarly activates neutrophils and contributes to their persistence in the asthmatic airways in spite of glucocorticoid therapy.
    Full-text · Article · May 2016
    • IL-8 is the predominant chemokine for neutrophils and was shown to be increased in BALF within hours after changing roughage feed from grass silage to hay in RAO affected horses [60]. In chronic inflammation IL-17 [61] and IL-1í µí»½ and IL-23 also play a role [62]. While the percentage of neutrophils in BALF decreases to reference levels in phases of remission, which hinders cytological diagnosis of the disease, myeloperoxidase (MPO) concentration in BALF was significantly higher in severe equine asthma during either crisis or remission compared to control horses [63].
    [Show abstract] [Hide abstract] ABSTRACT: Equine inflammatory airway disease (IAD) and recurrent airway obstruction (RAO) represent a spectrum of chronic inflammatory disease of the airways in horses resembling human asthma in many aspects. Therefore, both are now described as severity grades of equine asthma. Increasing evidence in horses and humans suggests that local pulmonary inflammation is influenced by systemic inflammatory processes and the other way around. Inflammation, coagulation, and fibrinolysis as well as extracellular remodeling show close interactions. Cytology of bronchoalveolar lavage fluid and tracheal wash is commonly used to evaluate the severity of local inflammation in the lung. Other mediators of inflammation, like interleukins involved in the chemotaxis of neutrophils, have been studied. Chronic obstructive pneumopathies lead to remodeling of bronchial walls and lung parenchyma, ultimately causing fibrosis. Matrix metalloproteinases (MMPs) are discussed as the most important proteolytic enzymes during remodeling in human medicine and increasing evidence exists for the horse as well. A systemic involvement has been shown for severe equine asthma by increased acute phase proteins like serum amyloid A and haptoglobin in peripheral blood during exacerbation. Studies focusing on these and further possible inflammatory markers for chronic respiratory disease in the horse are discussed in this review of the literature.
    Full-text · Article · Jan 2016
    • Other studies, however, suggest that a T H 1 response and cytokines (e.g. IL-8, IL-17) are responsible for neutrophil recruitment in RAO[10][11][12][13][14][15][16]. A study performed with horses affected by summer pasture-associated obstructive pulmonary disease (SPAOPD) revealed that the expression * Correspondence: couetill@purdue.edu 2 Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, 625 Harrison Street, West Lafayette, Indiana, 47907, USA Full list of author information is available at the end of the article of T H 1 and T H 2 cytokines varies throughout the year[17].
    [Show abstract] [Hide abstract] ABSTRACT: Recurrent airway obstruction (RAO) is a severe chronic respiratory disease affecting horses worldwide, though mostly in the Northern hemisphere. Environmental as well as genetic factors strongly influence the course and prognosis of the disease. Research has been focused on characterization of immunologic factors contributing to inflammatory responses, on genetic linkage analysis, and, more recently, on proteomic analysis of airway secretions from affected horses. The goal of this study was to investigate the interactions between eight candidate genes previously identified in a genetic linkage study and proteins expressed in bronchoalveolar lavage fluid (BALF) collected from healthy and RAO-affected horses. The analysis was carried out with Ingenuity Pathway Analysis® bioinformatics software. The gene with the greatest number of indirect interactions with the set of proteins identified is Interleukin 4 Receptor (IL-4R), whose protein has also been detected in BALF. Interleukin 21 receptor and chemokine (C-C motif) ligand 24 also showed a large number of interactions with the group of detected proteins. Protein products of other genes like that of SOCS5, revealed direct interactions with the IL-4R protein. The interacting proteins NOD2, RPS6KA5 and FOXP3 found in several pathways are reported regulators of the NFκB pathway. The pathways generated with IL-4R highlight possible important intracellular signaling cascades implicating, for instance, NFκB. Furthermore, the proposed interaction between SOCS5 and IL-4R could explain how different genes can lead to identical clinical RAO phenotypes, as observed in two Swiss Warmblood half sibling families because these proteins interact upstream of an important cascade where they may act as a functional unit.
    Full-text · Article · Aug 2011
    • Further, Laan et al. (2005) reported an increased production of IL-1 and TNFprotein by alveolar macrophages after exposure to HDS or AF. Conversely, Ainsworth et al. (2007) found in vitro exposure to hay dust did not result in increased expression of IL-1 mRNA by pulmonary mononuclear cells isolated from horses with RAO compared to control horses. In other studies, treatment of RAO-affected horses with clenbuterol or fluticasone proprionate has been shown to result in reduced expression of IL-1 and/or TNF-mRNA in BALF (Laan et al., 2006b; DeLuca et al., 2008), consistent with a role of these cytokines in airway inflammation.
    [Show abstract] [Hide abstract] ABSTRACT: Inflammatory airway disease (IAD) is a common disorder of performance horses and is associated with poor performance and accumulation of mucus and inflammatory cells in lower airway secretions. Horses with IAD frequently have increased relative counts of neutrophils in bronchoalveolar lavage fluid (BALF); less commonly relative counts of eosinophils and/or mast cells may be increased. The aetiopathogenesis of IAD is unknown and may involve innate and/or acquired immune responses to various factors including respirable dust constituents, micro-organisms, noxious gases and unconditioned air. The molecular pathways and role of the immune system in the pathogenesis of IAD remain poorly defined and it is unknown whether polarised T cell responses occur in the disease, as have been reported to occur in equine recurrent airway obstruction and asthma in humans. Elucidating cytokine responses that develop in horses with IAD may allow a greater understanding of the possible aetiopathological pathway(s) involved and could contribute to development of novel treatments. We compared the mRNA expression of tumour necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-8, IL-13, IL-17 and IL-23 in cell pellets extracted from BALF of horses with IAD (n=21) and horses free of respiratory tract disease (n=17). Horses with IAD had significantly increased levels of TNF-α, IL-1β and IL-23 mRNA; no significant differences in the other cytokine mRNAs were detected. The results of this study indicate that IAD of horses is associated with increased mRNA expression of pro-inflammatory cytokines in BALF cells, which may reflect stimulation of the innate immune responses to inhaled antigens. There was no evidence of a polarised T-cell cytokine response suggesting hypersensitivity responses may not be involved in the aetiopathogenesis of IAD.
    Full-text · Article · Mar 2011
    • Laan et al. (2006) found significant increase of IL-1β mRNA in equine pulmonary macrophages , but not constantly, at 6 and 24 hours when stimulated by Escherichia coli derived lypopolysaccharide or hay dust suspension. However in another study, it has been evidenced that IL-1β mRNA from pulmonary mononuclear cells isolated from RAO-affected horses exposed to dusty hay was not greater than in healthy horses (Ainsworth et al. 2007 ). On the other hand, equine bronchi passively sensitized with serum from animal suffering from RAO, showed a significant increase in IL-1β mRNA (Matera et al. 2005).
    [Show abstract] [Hide abstract] ABSTRACT: During an 18 day test, we measured the cytokine mRNA expression (Interleukin-1beta [IL-1beta], Interleukin-8 [IL-8], Interferon-gamma [IFN-gamma], Tumor Necrosis Factor-alpha [TNF-alpha]) of cells from bronchoalveolar lavage fluid [BALF] in five horses previously diagnosed with RAO, before and during challenge exposure, and after the desensitization phase which involved dexamethasone treatment and environmental modification. Simultaneously, the same cytokine mRNA expression of cells from BALF in four asymptomatic RAO-affected horses maintained outdoors was analyzed. An evident respiratory distress was observed in the challenge group within 3 days, with a significant overexpression of IL-8 and TNF-alpha mRNA on the ninth day. The pharmacological and environmental desensitization provided a down regulation of all the cytokines. No statistical modification characterized the cytokine kinetics of the asymptomatic horses maintained outdoors. A comparison for each time point of the cytokines between the exposed and unexposed horses showed no significant differences. The study suggested that a standardized exposure protocol and sampling time in experimental studies of RAO is mandatory for a correct comparison of the results obtained by different Authors. However, the absence of significant changes between the exposed and unexposed horses could depend on the lack of the sample uniformity since the evolution of the disease represents a continuum from a healthy to a pathological condition.
    Full-text · Article · Jan 2011
    • As mentioned above, IgE-mediated type I reactions and type III reactions have been suggested to be involved in the pathogenesis of heaves (Halliwell et al 1993, Schmallenbach et al 1998, Eder et al 2000, Franchini et al 2000, Eder et al 2001, Lavoie et al 2001, Curik et al 2003, Künzle et al 2007, Tahon et al 2009, Moran et al 2010 ). Moreover, recent findings suggest that in addition to the contribution of inflammatory cytokines, such as IL-1β, IL-8 and IL-17 (Cordeau et al 2004, Ainsworth et al 2007, Pietra et al 2007, Riihimäki et al 2008), helper T lymphocytes may contribute to the pathogenesis of heaves through their secretion of either Th1 or Th2 cytokines (Giguere et al 2002, Ainsworth et al 2003, Horohov et al 2005). Resident lung cells have also been implicated in the onset of the airway inflammatory response against A. fumigatus.
    [Show abstract] [Hide abstract] ABSTRACT: El objetivo de este estudio fue determinar si ratones expuestos a esporas de Aspergillus fumigatus desarrollan una inflamación alérgica de las vías respiratorias similar a la observada en equinos con Obstrucción Recurrente de las Vías Aéreas (ORVA). Para esto se utilizaron ratones Rockefeller (RK) de 2, 5, 6 y 8 meses respectivamente, los cuales se mantuvieron por 16 días en jaulas que contenían heno contaminado con A. fumigatus. Posterior a la exposición, los animales fueron sangrados y sacrificados para la obtención de suero sanguíneo, lavado broncoalveolar (LBA) y muestras de tejido pulmonar. Además, se midieron los niveles de anticuerpos específicos para A. fumigatus a través de un ELISA indirecto. Los resultados mostraron que la inhalación de esporas de A. fumigatus en ratones induce una respuesta alérgica inflamatoria en pulmones similar a lo observado en ORVA equino, con un aumento del porcentaje de neutrófilos en el LBA y cambios histológicos pulmonares característicos de una hipersensibilidad bronquial. Asimismo, se observó un aumento de los niveles de inmunoglubulinas IgE, IgG1 e IgG2a específicas para este hongo en fluidos bronquiales. Además, los animales remitieron el estrés respiratorio cuando fueron expuestos en un ambiente de remisión. Se puede concluir que la simple exposición de esporas de A. fumugatus produce una condición alérgica similar al ORVA en equinos. Por lo tanto, este modelo experimental puede ser de utilidad para el estudio de los mecanismos inmunológicos, y probar nuevas opciones terapéuticas para esta enfermedad respiratoria de los caballos.
    Full-text · Article · Dec 2010
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