[en] BACKGROUND: The aim of this study was to evaluate the contribution of bronchial epithelium to airway inflammation, with focus on mRNA and protein expression of cytokines of innate immunity IL-6, IL-10 and TNF-alpha, in horses with Recurrent Airway Obstruction (RAO) during exacerbation and in remission. RESULTS: Despite marked clinical and physiologic alterations between exacerbation and after remission in the RAO horses no differences were detected in either cytokine mRNA or protein levels. Moreover, the expression of investigated cytokines in RAO horses on pasture did not differ from controls.In comparing real-time PCR analysis to results of immunohistochemistry only IL-10 mRNA and protein levels in RAO horses on pasture were significantly correlated (rs = 0.893, p = 0.007). Curiously, in controls examined on pasture the TNF-alpha protein level was positively correlated to IL-10 mRNA expression (rs = 0.967, p = 0.007) and negatively correlated to IL-6 mRNA expression (rs = -0.971, p = 0.001). CONCLUSION: Given the complementary relationship of assessing cytokines directly by immunohistochemistry, or indirectly by PCR to mRNA, the lack of significant changes in either mRNA or protein levels of IL-6, IL-10 or TNF-alpha mRNA in RAO horses in exacerbation suggests that these particular cytokines in bronchial tissue may not play a substantive role in the active inflammation of this disease. To support this contention further studies examining time dependency of expression of IL-6, IL-10 or TNF-alpha are needed, as is expansion of the range of cytokines to include other key regulators of airway inflammation.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Riihimäki, Miia; Swedish University of Agricultural Sciences - SLU > Clinical sciences, Equine internal medicine
Raine, Amanda; Biomedical Centre Uppsala, Sweden > Cell and Molecular biology
Pourazar, Jamshid; University Hospital, Umea, Sweden > Respiratory Medicine and Allergy
Sandström, Thomas; University Hospital, Umea, Sweden > Respiratory Medicine and Allergy
Art, Tatiana ; Université de Liège - ULiège > Faculty of Veterinary Medicine, Equine Sports Medicine Centre
Lekeux, Pierre ; Université de Liège - ULiège > Faculty of Veterinary Medicine, Equine Sports Medicine Centre
Couëtil, Laurent; Purdue University, West Lafayette, IN, USA > Veterinary clinical sciences, School of veterinary medicine
Pringle, John; Swedish University of Agricultural Sciences - SLU > Clinical sciences, Equine internal medicine
Language :
English
Title :
Epithelial expression of mRNA and protein for IL-6, IL-10 and TNF-alpha in endobronchial biopsies in horses with recurrent airway obstruction.
Knight DA, Holgate ST: The airway epithelium: structural and functional properties in health and disease. Respirology 2003, 8(4):432-446.
Takizawa H: Bronchial epithelial cells in allergic reactions. Curr Drug Targets Inflamm Allergy 2005, 4(3):305-311.
Takizawa H: Airway epithelial cells as regulators of airway inflammation (Review). Int J Mol Med 1998, 1(2):367-378.
Holgate ST, Lackie P, Wilson S, Roche W, Davies D: Bronchial epithelium as a key regulator of airway allergen sensitization and remodeling in asthma. Am J Respir Crit Care Med 2000, 162 (3 Pt 2):S113-7.
Holgate ST: Inflammatory and structural changes in the airways of patients with asthma. Respir Med 2000, 94 Suppl D:S3-6.
Pepe C, Foley S, Shannon J, Lemiere C, Olivenstein R, Ernst P, Ludwig MS, Martin JG, Hamid Q: Differences in airway remodeling between subjects with severe and moderate asthma. J Allergy Clin Immunol 2005, 116(3):544-549.
Brown JL, Behndig AF, Sekerel BE, Pourazar J, Blomberg A, Kelly FJ, Sandstrom T, Frew AJ, Wilson SJ: Lower airways inflammation in allergic rhinitics: a comparison with asthmatics and normal controls. Clin Exp Allergy 2007, 37(5):688-695.
Pourazar J, Mudway IS, Samet JM, Helleday R, Blomberg A, Wilson SJ, Frew AJ, Kelly FJ, Sandstrom T: Diesel exhaust activates redoxsensitive transcription factors and kinases in human airways. Am J Physiol Lung Cell Mol Physiol 2005, 289(5):L724-30.
Wallin A, Sue-Chu M, Bjermer L, Ward J, Sandstrom T, Lindberg A, Lundback B, Djukanovic R, Holgate S, Wilson S: Effect of inhaled fluticasone with and without salmeterol on airway inflammation in asthma. J Allergy Clin Immunol 2003, 112(1):72-78.
Jeffery P, Holgate S, Wenzel S: Methods for the assessment of endobronchial biopsies in clinical research: application to studies of pathogenesis and the effects of treatment. Am J Respir Crit Care Med 2003, 168 (6 Pt 2):S1-17.
Howarth PH, Bradding P, Montefort S, Peroni D, Djukanovic R, Carroll MP, Holgate ST: Mucosal inflammation and asthma. Am J Respir Crit Care Med 1994, 150 (5 Pt 2):S18-22.
Ainsworth DM, Grunig G, Matychak MB, Young J, Wagner B, Erb HN, Antczak DF: Recurrent airway obstruction (RAO) in horses is characterized by IFN-gamma and IL-8 production in bronchoalveolar lavage cells. Veterinary immunology and immunopathology 2003, 96 (1-2):83-91.
Ainsworth DM, Wagner B, Franchini M, Grunig G, Erb HN, Tan JY: Time-dependent alterations in gene expression of interleukin-8 in the bronchial epithelium of horses with recurrent airway obstruction. Am J Vet Res 2006, 67(4):669-677.
Davis MS, Malayer JR, Vandeventer L, Royer CM, McKenzie EC, Williamson KK: Cold weather exercise and airway cytokine expression. J Appl Physiol 2005, 98(6):2132-2136.
Davis MS, Williams CC, Meinkoth JH, Malayer JR, Royer CM, Williamson KK, McKenzie EC: Influx of neutrophils and persistence of cytokine expression in airways of horses after performing exercise while breathing cold air. Am J Vet Res 2007, 68(2):185-189.
Berndt A, Derksen FJ, Venta P, Ewart S, Yuzbasiyan-Gurkan V, Robinson NE: Elevated amount of toll-like receptor 4 mRNA in bronchial epithelial cells is associated with airway inflammation in horses with Recurrent Airway Obstruction. Am J Physiol Lung Cell Mol Physiol 2006.
Debrue M, Hamilton E, Joubert P, Lajoie-Kadoch S, Lavoie JP: Chronic exacerbation of equine heaves is associated with an increased expression of interleukin-17 mRNA in bronchoalveolar lavage cells. Veterinary immunology and immunopathology 2005, 105 (1-2):25-31.
Gerber V, Robinson NE, Venta RJ, Rawson J, Jefcoat AM, Hotchkiss JA: Mucin genes in horse airways: MUC5AC, but not MUC2, may play a role in recurrent airway obstruction. Equine Vet J 2003, 35(3):252- 257.
Laan TT, Bull S, Pirie R, Fink-Gremmels J: The role of alveolar macrophages in the pathogenesis of recurrent airway obstruction in horses. J Vet Intern Med 2006, 20(1):167-174.
Giguere S, Viel L, Lee E, MacKay RJ, Hernandez J, Franchini M: Cytokine induction in pulmonary airways of horses with heaves and effect of therapy with inhaled fluticasone propionate. Veterinary immunology and immunopathology 2002, 85 (3-4):147-158.
van den Hoven R, Duvigneau JC, Hartl RT, Gemeiner M: Clenbuterol affects the expression of messenger RNA for interleukin 10 in peripheral leukocytes from horses challenged intrabronchially with lipopolysaccharides. Vet Res Commun 2006, 30(8):921-928.
Wang Z, Malmberg P, Larsson P, Larsson BM, Larsson K: Time course of interleukin-6 and tumor necrosis factor-alpha increase in serum following inhalation of swine dust. Am J Respir Crit Care Med 1996, 153(1):147-152.
de Waal Malefyt R, Abrams J, Bennett B, Figdor CG, de Vries JE: Interleukin 10 (IL-10) inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes. J Exp Med 1991, 174(5):1209-1220.
Gamble E, Qiu Y, Wang D, Zhu J, Vignola AM, Kroegel C, Morell F, Hansel TT, Pavord ID, Rabe KF, Barnes NC, Jeffery PK: Variability of bronchial inflammation in chronic obstructive pulmonary disease: implications for study design. Eur Respir J 2006, 27(2):293-299.
Haley KJ, Sunday ME, Wiggs BR, Kozakewich HP, Reilly JJ, Mentzer SJ, Sugarbaker DJ, Doerschuk CM, Drazen JM: Inflammatory cell distribution within and along asthmatic airways. Am J Respir Crit Care Med 1998, 158(2):565-572.
Art T, Franck T, Lekeux P, de Moffarts B, Couetil L, Becker M, Kohnen S, Deby-Dupont G, Serteyn D: Myeloperoxidase concentration in bronchoalveolar lavage fluid from healthy horses and those with recurrent airway obstruction. Can J Vet Res 2006, 70(4):291-296.
Couetil LL, Art T, de Moffarts B, Becker M, Melotte D, Jaspar F, Bureau F, Lekeux P: DNA binding activity of transcription factors in bronchial cells of horses with recurrent airway obstruction. Veterinary immunology and immunopathology 2006, 113 (1-2):11-20.
Riihimaki M, Raine A, Art T, Lekeux P, Couetil L, Pringle J: Partial divergence of cytokine mRNA expression in bronchial tissues compared to bronchoalveolar lavage cells in horses with recurrent airway obstruction. Veterinary immunology and immunopathology 2007.
You L, Yin J: Patterns of regulation from mRNA and protein time series. Metab Eng 2000, 2(3):210-217.
Britten KM, Howarth PH, Roche WR: Immunohistochemistry on resin sections: a comparison of resin embedding techniques for small mucosal biopsies. Biotech Histochem 1993, 68(5):271-280.
Couetil LL, Art T, de Moffarts B, Becker M, Melotte D, Jaspar F, Bureau F, Lekeux P: Effect of beclomethasone dipropionate and dexamethasone isonicotinate on lung function, bronchoalveolar lavage fluid cytology, and transcription factor expression in airways of horses with recurrent airway obstruction. J Vet Intern Med 2006, 20(2):399-406.
