We recently showed that serine proteases in German cockroach (GC) feces (frass) decreased experimental asthma through the activation of protease-activated receptor (PAR)-2. Since dendritic cells (DCs) play an important role in the initiation of asthma, we queried the role of GC frass proteases in modulating CCL20 (chemokine C-C motif ligand 20) and granulocyte macrophage colony-stimulating factor (GM-CSF) production, factors that regulate pulmonary DCs. A single exposure to GC frass resulted in a rapid, but transient, increase in GM-CSF and a steady increase in CCL20 in the airways of mice. Instillation of protease-depleted GC frass or instillation of GC frass in PAR-2-deficient mice significantly decreased chemokine release. A specific PAR-2-activating peptide was also sufficient to induce CCL20 production. To directly assess the role of the GC frass protease in chemokine release, we enriched the protease from GC frass and confirmed that the protease was sufficient to induce both GM-CSF and CCL20 production in vivo. Primary airway epithelial cells produced both GM-CSF and CCL20 in a protease- and PAR-2-dependent manner. Finally, we show a decreased percentage of myeloid DCs in the lung following allergen exposure in PAR-2-deficient mice compared to wild-type mice. However, there was no difference in GC frass uptake. Our data indicate that, through the activation of PAR-2, allergen-derived proteases are sufficient to induce CCL20 and GM-CSF production in the airways. This leads to increased recruitment and/or differentiation of myeloid DC populations in the lungs and likely plays an important role in the initiation of allergic airway responses.
© 2011 S. Karger AG, Basel
- Immune response
- Dendritic cell
- Asokananthan N, Graham PT, Stewart DJ, Bakker AJ, Eidne KA, Thompson PJ, Stewart GA: House dust mite allergens induce proinflammatory cytokines from respiratory epithelial cells: the cysteine protease allergen, Der p 1, activates protease-activated receptor (PAR)-2 and inactivates PAR-1. J Immunol 2002;169:4572–4578.
- Chiu LL, Perng DW, Yu CH, Su SN, Chow LP: Mold allergen, Pen c 13, induced IL-8 expression in human airway epithelial cells by activated protease-activated receptor 1 and 2. J Immunol 2007;178:5237–5244.
- Hong JH, Lee SI, Kim KE, Yong TS, Seo JT, Sohn MH, Shin DM: German cockroach extract activates protease-activated receptor 2 in human airway epithelial cells. J Allergy Clin Immunol 2004;113:315–319.
- Scarborough RM, Naughton MA, Teng W, Hung DT, Rose J, Vu TK, Wheaton VI, Turck CW, Coughlin SR: Tethered ligand agonist peptides. Structural requirements for thrombin receptor activation reveal mechanism of proteolytic unmasking of agonist function. J Biol Chem 1992;267:13146–13149.
- Colognato R, Slupsky JR, Jendrach M, Burysek L, Syrovets T, Simmet T: Differential expression and regulation of protease-activated receptors in human peripheral monocytes and monocyte-derived antigen-presenting cells. Blood 2003;102:2645–2652.
- Howells GL, Macey MG, Chinni C, Hou L, Fox MT, Harriott P, Stone SR: Proteinase-activated receptor-2: expression by human neutrophils. J Cell Sci 1997;110:881–887.
- D’Andrea MR, Rogahn CJ, Andrade-Gordon P: Localization of protease-activated receptors-1 and -2 in human mast cells: indications for an amplified mast cell degranuation cascade. Biotech Histochem 2000;75:85–90.
- Page K, Lierl K, Herman N, Wills-Karp M: Differences in susceptibility to German cockroach frass and its associated proteases in induced allergic inflammation in mice. Respir Res 2007;8:91.
- Page K, Ledford JR, Zhou P, Wills-Karp M: Mucosal sensitization to German cockroach involves protease-activated receptor-2. Respir Res 2010;11:62.
- Jahnsen FL, Strickland DH, Thomas JA, Tobagus IT, Napoli S, Zosky GR, Turner DJ, Sly PD, Stumbles PA, Holt PG: Accelerated antigen sampling and transport by airway mucosal dendritic cells following inhalation of a bacterial stimulus. J Immunol 2006;177:5861–5867.
- Jahnsen FL, Moloney ED, Hogan T, Upham JW, Burke CM, Holt PG: Rapid dendritic cell recruitment to the bronchial mucosa of patients with atopic asthma in response to local allergen challenge. Thorax 2001;56:823–826.
- Weckmann M, Collison A, Simpson JL, Kopp MV, Wark PA, Smyth MJ, Yagita H, Matthaei KI, Hansbro N, Whitehead B, Gibson PG, Foster PS, Mattes J: Critical link between TRAIL and CCL20 for the activation of Th2 cells and the expression of allergic airway disease. Nature Med 2007;13:1308–1315.
- Dieu MC, Vanbervliet B, Vicari A, Bridon JM, Oldham E, Aït-Yahia S, Brière F, Zlotnik A, Lebecque S, Caux C: Selective recruitment of immature and mature dendritic cells by distinct chemokines expressed in different anatomic sites. J Exp Med 1998;188:373–386.
- Nathan AT, Peterson EA, Chakir J, Wills-Karp M: Innate immune responses of airway epithelium to house dust mite are mediated through β-glucan-dependent pathways. J Allergy Clin Immunol 2009;123:612–618.
- Esashi E, Wang YH, Perng O, Qin XF, Liu YJ, Watowich SS: The signal transducer STAT5 inhibits plasmacytoid dendritic cell development by suppressing transcription factor IRF8. Immunity 2008;28:509–520.
- de Heer HJ, Hammad H, Soullié T, Hijdra D, Vos N, Willart MA, Hoogsteden HC, Lambrecht BN: Essential role of lung plasmacytoid dendritic cells in preventing asthmatic reactions to harmless inhaled antigen. J Exp Med 2004;200:89–98.
- Cates EC, Fattouh R, Wattie J, Inman MD, Goncharova S, Coyle AJ, Gutierrez-Ramos JC, Jordana M: Intranasal exposure of mice to house dust mite elicits allergic airway inflammation via a GM-CSF-mediated mechanism. J Immunol 2004;173:6384–6392.
- Stämpfli MR, Wiley RE, Neigh GS, Gajewska BU, Lei XF, Snider DP, Xing Z, Jordana M: Gm-Csf transgene expression in the airway allows aerosolized ovalbumin to induce allergic sensitization in mice. J Clin Invest 1998;102:1704–1714.
- Chavira RCJ, Burnett TJ, Hageman JH: Assaying proteinases with azocoll. Anal Biochem 1984;136:446–450.
- Bhat RK, Page K, Tan A, Hershenson MB: German cockroach extract increases bronchial epithelial cell interleukin-8 expression. Clin Exp Allergy 2003;33:35–42.
- Hughes VS, Page K: German cockroach frass proteases cleave pro-matrix metalloproteinase-9. Exp Lung Res 2007;33:135–150.
- Walters DM, Breysse PN, Wills-Karp M: Ambient urban Baltimore particulate-induced airway hyperresponsiveness and inflammation in mice. Am J Respir Crit Care Med 2001;164:1438–1443.
- Page K, Hughes VS, Bennett GW, Wong HR: German cockroach proteases regulate matrix metalloproteinase-9 in human bronchial epithelial cells. Allergy 2006;61:988–995.
- Page K, Lierl KM, Hughes VS, Zhou P, Ledford JR, Wills-Karp M: TLR2-mediated activation of neutrophils in response to German cockroach frass. J Immunol 2008;180:6317–6324.
- Vermaelen KY, Carro-Muino I, Lambrecht BN, Pauwels RA: Specific migratory dendritic cells rapidly transport antigen from the airways to the thoracic lymph nodes. J Exp Med 2001;193:51–60.
- Page K, Hughes VS, Odoms KK, Dunsmore KE, Hershenson MB: German cockroach proteases regulate IL-8 expression via NF-IL6 in human bronchial epithelial cells. Am J Respir Cell Mol Biol 2005;32:225–231.
- Page K, Strunk VS, Hershenson MB: Cockroach proteases increase IL-8 expression in human bronchial epithelial cells via activation of protease-activated receptor (PAR)-2 and ERK. J Allergy Clin Immunol 2003;112:1112–1118.
- Pichavant M, Charbonnier AS, Taront S, Brichet A, Wallaert B, Pestel J, Tonnel AB, Gosset P: Asthmatic bronchial epithelium activated by the proteolytic allergen Der p 1 increases selective dendritic cell recruitment. J Allergy Clin Immunol 2005;115:771–778.
- Rohani MG, Beyer RP, Hacker BM, Dommisch H, Dale BA, Chung WO: Modulation of expression of innate immunity markers CXCL5/ENA-78 and CCL20/MIP3α by protease-activated receptors (PARs) in human gingival epithelial cells. Innate Immun 2010;16:104–114.
- Fields RC, Schoenecker JG, Hart JP, Hoffman MR, Pizzo SV, Lawson JH: Protease-activated receptor-2 signaling triggers dendritic cell development. Am J Path 2003;162:1817–1822.
- Ramelli G, Fuertes S, Busso N, Acha-Orbea H, So A: Protease-activated receptor 2 signalling promotes dendritic cell antigen transport and T-cell activation in vivo. Immunol 2010;129:20–27.
- King C, Brennan S, Thompson PJ, Stewart GA: Dust mite proteolytic allergens induce cytokine release from cultured airway epithelium. J Immunol 1998;161:3645–3651.
- Sun G, Stacey MA, Schmidt M, Mori L, Mattoli S: Interaction of mite allergens Der p 3 and Der p 9 with protease-activated receptor-2 expressed by lung epithelial cells. J Immunol 2001;167:1014–1021.
- Ogawa T, Takai T, Kato T, Kikuchi Y, Niyonsaba F, Ikeda S, Okumura K, Ogawa H: Upregulation of the release of granulocyte-macrophage colony-stimulating factor from keratinocytes stimulated with cysteine protease activity of recombinant major mite allergens, Der f 1 and Der p 1. Int Arch Allergy Immunol 2008;146:27–35.
- Lordan JL, Bucchieri F, Richter A, Konstantinidis A, J.W. H, Thornber M, Puddicombe SM, Buchanan D, Wilson SJ, Djukanovic R, Holgate ST, Davies DE: Cooperative effects of Th2 cytokines and allergen on normal and asthmatic bronchial epithelial cells. J Immunol 2002;169:407–414.
- Rudack C, Steinhoff M, Mooren F, Becker K, von Eiff C, Sachse F: PAR-2 activation regulastes IL-8 and GRO-α synthesis by NF-ĸB, but not RANTES, IL-6, eotaxin, or TARC expression in nasal epithelium. Clin Exp Allergy 2007;37:1009–1022.
Dr. Kristen Page
Division of Critical Care Medicine
Cincinnati Children’s Hospital Medical Center
3333 Burnet Ave, ML 7006, Cincinnati, OH 45229 (USA)
Tel. +1 513 636 3079, E-Mail firstname.lastname@example.org
Received: March 4, 2011
Accepted after revision: May 8, 2011
Published online: August 29, 2011
Number of Print Pages : 11
Number of Figures : 6, Number of Tables : 1, Number of References : 36
Journal of Innate Immunity
Vol. 4, No. 1, Year 2012 (Cover Date: December 2011)
Journal Editor: Herwald H. (Lund), Egesten A. (Lund)
ISSN: 1662-811X (Print), eISSN: 1662-8128 (Online)
For additional information: http://www.karger.com/JIN
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.