Inhibition of neutrophil elastase attenuates airway hyperresponsiveness and inflammation in a mouse model of secondary allergen challenge: neutrophil elastase inhibition attenuates allergic airway responses

Respiratory Research

Arihiko Kanehiro²  Mitsune Tanimoto²  Erwin W Gelfand¹  Mikio Kataoka²  Yasushi Tanimoto²  Katsuichiro Ono²  Koichi Waseda²  Genyo Ikeda²  Yasuko Fuchimoto²  Nobuaki Miyahara²  Hikari Koga² 

[1] Department of Pediatrics, Division of Cell Biology, National Jewish Health, Denver, Colorado, USA;Department of Hematology, Oncology, Allergy and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan

关键词: Asthma;    Hyperresponsiveness;    Airway;    Elastase;    Neutrophil;   

Others  :  796551
DOI  :  10.1186/1465-9921-14-8

 received in 2012-10-30, accepted in 2013-01-17,  发布年份 2013

【 摘 要 】

Background

Chronic asthma is often associated with neutrophilic infiltration in the airways. Neutrophils contain elastase, a potent secretagogue in the airways, nonetheless the role for neutrophil elastase as well as neutrophilic inflammation in allergen-induced airway responses is not well defined. In this study, we have investigated the impact of neutrophil elastase inhibition on the development of allergic airway inflammation and airway hyperresponsiveness (AHR) in previously sensitized and challenged mice.

Methods

BALB/c mice were sensitized and challenged (primary) with ovalbumin (OVA). Six weeks later, a single OVA aerosol (secondary challenge) was delivered and airway inflammation and airway responses were monitored 6 and 48 hrs later. An inhibitor of neutrophil elastase was administered prior to secondary challenge.

Results

Mice developed a two-phase airway inflammatory response after secondary allergen challenge, one neutrophilic at 6 hr and the other eosinophilic, at 48 hr. PAR-2 expression in the lung tissues was enhanced following secondary challenge, and that PAR-2 intracellular expression on peribronchial lymph node (PBLN) T cells was also increased following allergen challenge of sensitized mice. Inhibition of neutrophil elastase significantly attenuated AHR, goblet cell metaplasia, and inflammatory cell accumulation in the airways following secondary OVA challenge. Levels of IL-4, IL-5 and IL-13, and eotaxin in BAL fluid 6 hr after secondary allergen challenge were significantly suppressed by the treatment. At 48 hr, treatment with the neutrophil elastase inhibitor significantly reduced the levels of IL-13 and TGF-β1 in the BAL fluid. In parallel, in vitro IL-13 production was significantly inhibited in spleen cells from sensitized mice.

Conclusion

These data indicate that neutrophil elastase plays an important role in the development of allergic airway inflammation and hyperresponsiveness, and would suggest that the neutrophil elastase inhibitor reduced AHR to inhaled methacholine indicating the potential for its use as a modulator of the immune/inflammatory response in both the neutrophil- and eosinophil-dominant phases of the response to secondary allergen challenge.

【 授权许可】

   
2013 Koga et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Lee NA, Gelfand EW, Lee JJ: Pulmonary T cells and eosinophils: coconspirators or independent triggers of allergic respiratory pathology? J Allergy Clin Immunol 2001, 107:945-957.
• [2]Larché M, Robinson DS, Kay AB: The role of T lymphocytes in the pathogenesis of asthma. J Allergy Clin Immunol 2003, 111:450-463.
• [3]Simpson JL, Scott R, Boyle MJ, Gibson PG: Inflammatory subtypes in asthma: assessment and identification using induced sputum. Respirology 2006, 11:54-61.
• [4]Wenzel SE, Szefler SJ, Leung DY, Sloan SI, Rex MD, Martin RJ: Bronchoscopic evaluation of severe asthma. Persistent inflammation associated with high dose glucocorticoids. Am J Respir Crit Care Med 1997, 156:737-743.
• [5]Foley SC, Hamid Q: Images in allergy and immunology: neutrophils in asthma. J Allergy Clin Immunol 2007, 119:1282-1286.
• [6]Fahy JV: Eosinophilic and neutrophilic inflammation in asthma: insights from clinical studies. Proc Am Thorac Soc 2009, 6:256-259.
• [7]Lamblin C, Gosset P, Tillie-Leblond I, Saulnier F, Marquette CH, Wallaert B, Tonnel AB: Bronchial neutrophilia in patients with noninfectious status asthmaticus. Am J Respir Crit Care Med 1998, 157:394-402.
• [8]Kanehiro A, Ikemura T, Makela MJ, Lahn M, Joetham A, Dakhama A, Gelfand EW: Inhibition of phosphodiesterase 4 attenuates airway hyperresponsiveness and airway inflammation in a model of secondary allergen challenge. Am J Respir Crit Care Med 2001, 163:173-184.
• [9]Takeda K, Miyahara N, Kodama T, Taube C, Balhorn A, Dakhama A, Kitamura K, Hirano A, Tanimoto M, Gelfand EW: S-carboxymethylcysteine normalises airway responses in sensitized and challenged mice. Eur Respir J 2005, 26:577-585.
• [10]Suzuki T, Wang W, Lin JT, Shirato K, Mitsuhashi H, Inoue H: Aerosolized human neutrophil elastase induces airway constriction and hyperresponsiveness with protection by intravenous pretreatment with half-length secretory leukoprotease inhibitor. Am J Respir Crit Care Med 1996, 153:1405-1411.
• [11]Heutinck KM, ten Berge IJ, Hack CE, Hamann J, Rowshani AT: Serine proteases of the human immune system in health and disease. Mol Immunol 2010, 47:1943-1955.
• [12]Monteseirín J, Bonilla I, Camacho MJ, Chacón P, Vega A, Chaparro A, Conde J, Sobrino F: Specific allergens enhance elastase release in stimulated neutrophils from asthmatic patients. Int Arch Allergy Immunol 2003, 131:174-181.
• [13]Nyenhuis SM, Schwantes EA, Evans MD, Mathur SK: Airway neutrophil inflammatory phenotype in older subjects with asthma. J Allergy Clin Immunol 2010, 125:1163-1165.
• [14]Baines KJ, Simpson JL, Wood LG, Scott RJ, Gibson PG: Systemic upregulation of neutrophil α-defensins and serine proteases in neutrophilic asthma. Thorax 2011, 66:942-947.
• [15]Wood LG, Baines KJ, Fu J, Scott HA, Gibson PG: The neutrophilic inflammatory phenotype is associated with systemic inflammation in asthma. Chest 2012, 142:86-93.
• [16]Agusti C, Takeyama K, Cardell LO, Ueki I, Lausier J, Lou YP, Nadel JA: Goblet cell degranulation after antigen challenge in sensitized guinea pigs: role of neutrophils. Am J Respir Crit Care Med 1998, 158:1253-1258.
• [17]Nadel JA, Takeyama K, Agustí C: Role of neutrophil elastase in hypersecretion in asthma. Eur Respir J 1999, 13:190-196.
• [18]Fujimoto K, Kubo K, Shinozaki S, Okada K, Matsuzawa Y, Kobayashi T, Sugane K: Neutrophil elastase inhibitor reduces asthmatic responses in allergic sheep. Respir Physiol 1995, 100:91-100.
• [19]Fuchimoto Y, Kanehiro A, Miyahara N, Koga H, Ikeda G, Waseda K, Tanimoto Y, Ueha S, Kataoka M, Gelfand EW, Tanimoto M: Requirement for CCR5 in the development of allergen-induced airway hyperresponsiveness and inflammation. Am J Respir Cell Mol Biol 2011, 45:1248-1255.
• [20]Yashiro M, Tsukahara H, Matsukawa A, Yamada M, Fujii Y, Nagaoka Y, Tsuge M, Yamashita N, Ito T, Yamada M, Masutani H, Yodoi J, Morishima T: Redox-active protein thioredoxin-1 administration ameliorates influenza a virus (H1N1)-induced acute lung injury in mice. Crit Care Med 2013, 41:166-176.
• [21]Ito W, Kanehiro A, Matsumoto K, Hirano A, Ono K, Maruyama H, Kataoka M, Nakamura T, Gelfand EW, Tanimoto M: Hepatocyte growth factor attenuates airway hyperresponsiveness, inflammation, and remodeling. Am J Respir Cell Mol Biol 2005, 32:268-280.
• [22]Miyahara N, Takeda K, Miyahara S, Matsubara S, Koya T, Matsubara S, Joetham A, Krishnan E, Dakhama A, Haribabu B, Gelfand EW: Requirement for leukotriene B4 receptor 1 in allergen-induced airway hyperresponsiveness. Am J Respir Crit Care Med 2005, 172:161-167.
• [23]Miyahara N, Takeda K, Kodama T, Joetham A, Taube C, Park JW, Miyahara S, Balhorn A, Dakhama A, Gelfand EW: Contribution of antigen-primed CD8+ T cells to the development of airway hyperresponsiveness and inflammation is associated with IL-13. J Immunol 2004, 172:2549-2558.
• [24]Ito W, Tanimoto M, Ono K, Mizuno S, Yoshida A, Koga H, Fuchimoto Y, Kondo N, Tanimoto Y, Kiura K, Matsumoto K, Kataoka M, Nakamura T, Gelfand EW, Kanehiro A: Growth factors temporally associate with airway responsiveness and inflammation in allergen-exposed mice. Int Arch Allergy Immunol 2007, 145:324-339.
• [25]Uehara A, Muramoto K, Takada H, Sugawara S: Neutrophil serine proteinases activate human nonepithelial cells to produce inflammatory cytokines through protease-activated receptor 2. J Immunol 2003, 170:5690-5696.
• [26]Schmidlin F, Amadesi S, Vidil R, Trevisani M, Martinet N, Caughey G, Tognetto M, Cavallesco G, Mapp C, Geppetti P, Bunnett NW: Expression and function of proteinase-activated receptor 2 in human bronchial smooth muscle. Am J Respir Crit Care Med 2001, 164:1276-1281.
• [27]Cocks TM, Moffatt JD: Protease-activated receptor-2 (PAR-2) in the airways. Pulm Pharmacol Ther 2001, 14:183-191.
• [28]Bolton SJ, McNulty CA, Thomas RJ, Hewitt CR, Wardlaw AJ: Expression of and functional responses to protease-activated receptors on human eosinophils. J Leukoc Biol 2003, 74:60-68.
• [29]Rydell-Törmänen K, Johnson JR, Fattouh R, Jordana M, Erjefält JS: Induction of vascular remodeling in the lung by chronic house dust mite exposure. Am J Respir Cell Mol Biol 2008, 39:61-67.
• [30]Hammad H, Chieppa M, Perros F, Willart MA, Germain RN, Lambrecht BN: House dust mite allergen induces asthma via Toll-like receptor 4 triggering of airway structural cells. Nat Med 2009, 15:410-416.
• [31]Phipps S, Lam CE, Kaiko GE, Foo SY, Collison A, Mattes J, Barry J, Davidson S, Oreo K, Smith L, Mansell A, Matthaei KI, Foster PS: Toll/IL-1 signaling is critical for house dust mite-specific Th1 and Th2 responses. Am J Respir Crit Care Med 2009, 179:883-893.
• [32]Trompette A, Divanovic S, Visintin A, Blanchard C, Hegde RS, Madan R, Thorne PS, Wills-Karp M, Gioannini TL, Weiss JP, Karp CL: Allergenicity resulting from functional mimicry of a Toll-like receptor complex protein. Nature 2009, 457:585-588.
• [33]Ichikawa S, Takai T, Yashiki T, Takahashi S, Okumura K, Ogawa H, Kohda D, Hatanaka H: Lipopolysaccharide binding of the mite allergen Der f 2. Genes Cells 2009, 14:1055-1065.
• [34]Bobic S, Seys S, De Vooght V, Callebaut I, Hox V, Dooms C, Vinckier S, Jonckx B, Saint-Remy JM, Stassen JM, Bullens DM, Ceuppens JL, Carmeliet P, Hellings PW: Placental growth factor contributes to bronchial neutrophilic inflammation and edema in allergic asthma. Am J Respir Cell Mol Biol 2012, 46:781-789.
• [35]Nabe T, Ikedo A, Hosokawa F, Kishima M, Fujii M, Mizutani N, Yoshino S, Ishihara K, Akiba S, Chaplin DD: Regulatory role of antigen-induced interleukin-10, produced by CD4+ T cells, in airway neutrophilia in a murine model for asthma. Eur J Pharmacol 2012, 677:154-162.
• [36]Nocker RE, Out TA, Weller FR, Mul EP, Jansen HM, van der Zee JS: Influx of neutrophils into the airway lumen at 4 h after segmental allergen challenge in asthma. Int Arch Allergy Immunol 1999, 119:45-53.
• [37]Tomkinson A, Cieslewicz G, Duez C, Larson KA, Lee JJ, Gelfand EW: Temporal association between airway hyperresponsiveness and airway eosinophilia in ovalbumin-sensitized mice. Am J Respir Crit Care Med 2001, 163:721-730.
• [38]Macfarlane SR, Seatter MJ, Kanke T, Hunter GD, Plevin R: Protease-activated receptors. Pharmacol Rev 2001, 53:245-282.
• [39]Coughlin SR: Thrombin signalling and protease-activated receptors. Nature 2000, 407:258-264.
• [40]Hou L, Howells GL, Kapas S, Macey MG: The protease-activated receptors and their cellular expression and function in blood-related cells. Br J Haematol 1998, 101:1-9.
• [41]Schmidlin F, Amadesi S, Dabbagh K, Lewis DE, Knott P, Bunnett NW, Gater PR, Geppetti P, Bertrand C, Stevens ME: Protease-activated receptor 2 mediates eosinophil infiltration and hyperreactivity in allergic inflammation of the airway. J Immunol 2002, 169:5315-5321.
• [42]Hiraguchi Y, Nagao M, Hosoki K, Tokuda R, Fujisawa T: Neutrophil proteases activate eosinophil function in vitro. Int Arch Allergy Immunol 2008, 146:16-21.
• [43]Kikuchi I, Kikuchi S, Kobayashi T, Hagiwara K, Sakamoto Y, Kanazawa M, Nagata M: Eosinophil trans-basement membrane migration induced by interleukin-8 and neutrophils. Am J Respir Cell Mol Biol 2006, 34:760-765.
• [44]Ohnishi H, Miyahara N, Gelfand EW: The role of leukotriene B4 in allergic diseases. Allergol Int 2008, 57:291-298.
• [45]Waseda K, Miyahara N, Kanehiro A, Ikeda G, Koga H, Fuchimoto Y, Kurimoto E, Tanimoto Y, Kataoka M, Tanimoto M, Gelfand EW: Blocking the leukotriene B4 receptor 1 inhibits late phase airway responses in established disease. Am J Respir Cell Mol Biol 2011, 45:851-857.
• [46]Rothenberg ME, Luster AD, Leder P: Murine eotaxin: an eosinophil chemoattractant inducible in endothelial cells and in interleukin 4-induced tumor suppression. Proc Natl Acad Sci USA 1995, 92:8960-8964.
• [47]Lee CG, Homer RJ, Zhu Z, Lanone S, Wang X, Koteliansky V, Shipley JM, Gotwals P, Noble P, Chen Q, Senior RM, Elias JA: Interleukin-13 induces tissue fibrosis by selectively stimulating and activating transforming growth factor beta (1). J Exp Med 2001, 194:809-821.
• [48]Doucet C, Brouty-Boyé D, Pottin-Clémenceau C, Canonica GW, Jasmin C, Azzarone B: Interleukin (IL) 4 and IL-13 act on human lung fibroblasts. Implication in asthma. J Clin Invest 1998, 101:2129-2139.
• [49]Minshall EM, Leung DY, Martin RJ, Song YL, Cameron L, Ernst P, Hamid Q: Eosinophil-associated TGF-beta1 mRNA expression and airways fibrosis in bronchial asthma. Am J Respir Cell Mol Biol 1997, 17:326-333.
• [50]Hirano A, Kanehiro A, Ono K, Ito W, Yoshida A, Okada C, Nakashima H, Tanimoto Y, Kataoka M, Gelfand EW, Tanimoto M: Pirfenidone modulates airway responsiveness, inflammation and remodeling after repeated challenge. Am J Respir Cell Mol Biol 2006, 35:366-377.