Kinin Danger Signals Proteolytically Released by Gingipain Induce Fimbriae-Specific IFN-γ- and IL-17-Producing T Cells in Mice Infected This information is current as Intramucosally with Porphyromonas of September 26, 2021. gingivalis Ana Carolina Monteiro, Aline Scovino, Susane Raposo, Vinicius Mussa Gaze, Catia Cruz, Erik Svensjö, Marcelo Sampaio Narciso, Ana Paula Colombo, João B. Pesquero, Downloaded from Eduardo Feres-Filho, Ky-Anh Nguyen, Aneta Sroka, Jan Potempa and Julio Scharfstein J Immunol 2009; 183:3700-3711; Prepublished online 17 August 2009; doi: 10.4049/jimmunol.0900895 http://www.jimmunol.org/ http://www.jimmunol.org/content/183/6/3700 Supplementary http://www.jimmunol.org/content/suppl/2009/08/17/jimmunol.090089 Material 5.DC1 References This article cites 64 articles, 27 of which you can access for free at: by guest on September 26, 2021 http://www.jimmunol.org/content/183/6/3700.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2009 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Kinin Danger Signals Proteolytically Released by Gingipain Induce Fimbriae-Specific IFN-␥- and IL-17-Producing T Cells in Mice Infected Intramucosally with Porphyromonas gingivalis1 Ana Carolina Monteiro,* Aline Scovino,* Susane Raposo,*† Vinicius Mussa Gaze,*† Catia Cruz,* Erik Svensjo¨,* Marcelo Sampaio Narciso,‡ Ana Paula Colombo,§ Joa˜o B. Pesquero,¶ Eduardo Feres-Filho,† Ky-Anh Nguyen,ʈ# Aneta Sroka,** Jan Potempa,**†† and Julio Scharfstein2* Porphyromonas gingivalis, a Gram-negative bacterium that causes periodontitis, activates the kinin system via the cysteine pro- tease R-gingipain. Using a model of buccal infection based on P. gingivalis inoculation in the anterior mandibular vestibule, we Downloaded from studied whether kinins released by gingipain may link mucosal inflammation to T cell-dependent immunity through the activation of bradykinin B2 receptors (B2R). Our data show that P. gingivalis W83 (wild type), but not gingipain-deficient mutant or wild-type bacteria pretreated with gingipain inhibitors, elicited buccal edema and gingivitis in BALB/c or C57BL/6 mice. ؊/؊ ؊/؊ Studies in TLR2 ,B2R , and neutrophil-depleted C57BL/6 mice revealed that P. gingivalis induced edema through the sequential activation of TLR2/neutrophils, with the initial plasma leakage being amplified by gingipain-dependent release of vasoactive kinins from plasma-borne kininogens. We then used fimbriae (Fim) Ag as a readout to verify whether activation http://www.jimmunol.org/ 3 3 of the TLR2 PMN B2R axis (where PMN is polymorphonuclear neutrophil) at early stages of mucosal infection had impact ␥ on adaptive immunity. Analyzes of T cell recall responses indicated that gingipain drives B2R-dependent generation of IFN- - producing Fim T cells in submandibular draining lymph nodes of BALB/c and C57BL/6 mice, whereas IL-17-producing Fim T cells were generated only in BALB/c mice. In summary, our studies suggest that two virulence factors, LPS (an atypical TLR2 ligand) and gingipain, forge a trans-cellular cross-talk between TLR2 and B2R, thus forming an innate axis that guides the development of Fim-specific T cells in mice challenged intrabuccally by P. gingivalis. Ongoing research may clarify whether kinin-driven modulation of T cell responses may also influence the severity of chronic periodontitis. The Journal of Immunology, 2009, 183: 3700–3711. by guest on September 26, 2021 eriodontal disease is initiated by a chronic subgingival gingivalis, whereas the normal flora is dominated by facultative bacterial infection that destroys the underlying alveolar Gram-positive bacteria (1). Studies in laboratory animals infected P bone as well as the connective tissue that attaches the gin- with P. gingivalis (1) suggest that bacteria are required for the giva to the tooth root (1). During the progression of periodontitis, initiation of inflammation, whereas bone erosion is more likely composition of the subgingival biofilm is changed; there is an en- caused by aberrant immune responses to bacterial Ags persistent in richment of anaerobic Gram-negative bacteria belonging to the red the periodontal tissues. The latter hypothesis received indirect sup- complex group of periodontal pathogens, such as Porphyromonas port from pathogenesis studies in animal models of autoimmune arthritis, showing that activated T and B cells up-regulate RANKL (receptor activator of NF-␬B ligand), thus being capable of driving *Carlos Chagas Filho Institute of Biophysics, †Faculty of Odontology, ‡Department the differentiation of bone-erosive osteoclasts (2, 3). Efforts to of Histology, Institute of Biomedical Sciences, and §Pablo de Go´es Institute of Mi- crobiology, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; ¶Department characterize osteoclastogenic T cell subsets have recently con- ʈ of Biophysics, Federal University of Sa˜o Paulo, Sa˜o Paulo, Brazil; Institute of Dental verged at TH17 (4), a proinflammatory helper T cell subset whose Research, Westmead Millenium Institute and #Faculty of Dentistry, University of differentiation depends on the cooperative action of IL-6, TGF-␤, Sydney, Sydney, Australia; **Faculty of Biochemistry, Biophysics, and Biotechnol- ogy, Department of Microbiology, Jagiellonian University, Krakow, Poland; and and IL-23 (5–9). Although the severity of chronic human peri- ††Department of Periodontics, University of Louisville School of Dentistry, Louis- odontal disease is positively correlated with IL-17 levels (10–12), ville, KY 40202 the hypothesis that osteoclastogenic human TH17 cells have a Received for publication March 20, 2009. Accepted for publication July 8, 2009. causal role in alveolar bone erosion remains to be confirmed. The costs of publication of this article were defrayed in part by the payment of page As a recently recognized a risk factor in the development of charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. atherosclerosis, both in humans and in animal models (13, 14), P. gingivalis relies on multiple virulence factors, such as fimbriae 1 This work was supported by grants from Instituto Nacional de Pesquisa em Biologia 3 Estrutural e Bio-Imagem do Conselho Nacional de Desenvolvimento Científico e (Fim), LPS, cysteine proteases, and hemagglutinin, to colonize Tecnolo´gico), Fundac¸a˜o de Amparo a Pesquisa do Estado do Rio de Janeiro/Programa Pensa Rio, and Coordenac¸a˜o de Aperfeic¸oamento de Pessoal de Nível Superior, National Institutes of Health Grants DE 09761 and 1642/B/P01/2008/35, and a 3 Abbreviations used in this paper: Fim, fimbriae; BK, bradykinin; B2R, BK B2 re- grant from the Department of Scientific Research, Polish Ministry of Science and ceptor; DC, dendritic cell; HCP, hamster cheek pouch; HK, human kininogen; LBK, Education. lysyl-BK; LN, lymph node; NOD, nucleotide-binding oligomerization domain; p.i., 2 Address correspondence and reprint requests to Dr. Julio Scharfstein, Universidade postinfection; PMN, polymorphonuclear neutrophil; Rgps, arginine-specific gingi- Federal do Rio de Janeiro, Centro de Ciencias de Saude, Ilha do Fundao, Bloco D, pain; TAFI, thrombin-activatable fibrinolysis inhibitor; WT, wild type. Sala D 007, Rio de Janeiro, Rio de Janeiro 21944-900, Brazil. E-mail address: [email protected] Copyright © 2009 by The American Association of Immunologists, Inc. 0022-1767/09/$2.00 www.jimmunol.org/cgi/doi/10.4049/jimmunol.0900895 The Journal of Immunology 3701 and survive in oral mucosal tissues (15). Molecular studies led to Analysis of host resistance mechanisms in mice acutely infected the characterization of two major TLR2 ligands of P. gingivalis:an with T. cruzi (i.p. route) demonstrated that splenic DCs activated atypical LPS (16) and Fim, a multiunit complex composed of po- via the kinin/B2R pathway play a critical role in the generation of lymerized fimbrillin (FimA) and accessory proteins (FimCDE) immunoprotective IFN-␥-producing T cells (47). (17). More recently, analysis of macrophage responses to P. gin- For this report, we studied the impact of kinin system activation givalis or to its purified wall components (Fim and LPS) revealed on T cell development in mice challenged by a low intrabuccal that NF␬B-responsive genes were activated via TLR2- or TLR7- dose of P. gingivalis. Analysis of the dynamics of the inflamma- MyD88-p38 MAPK pathways, respectively (18). In another in tory response evoked by this periodontal pathogen revealed that vitro study, it was shown that Fim provide adhesive filamentous plasma leakage, a vascular response orchestrated by TLR2/neutro- appendages for gingival epithelial cells without triggering TLR2- phil activation at very early stages of infection, is a limiting step mediated