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Dependent Cathepsin S Expression Γ IFN IFN Regulatory Factor-1 Regulates IFN-γ -Dependent Cathepsin S Expression Karin Storm van's Gravesande, Matthew D. Layne, Qiang Ye, Louis Le, Rebecca M. Baron, Mark A. Perrella, Laura This information is current as Santambrogio, Eric S. Silverman and Richard J. Riese of September 26, 2021. J Immunol 2002; 168:4488-4494; ; doi: 10.4049/jimmunol.168.9.4488 http://www.jimmunol.org/content/168/9/4488 Downloaded from References This article cites 39 articles, 13 of which you can access for free at: http://www.jimmunol.org/content/168/9/4488.full#ref-list-1 http://www.jimmunol.org/ 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 by guest on September 26, 2021 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 © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. IFN Regulatory Factor-1 Regulates IFN-␥-Dependent Cathepsin S Expression1 Karin Storm van’s Gravesande,* Matthew D. Layne,* Qiang Ye,* Louis Le,* Rebecca M. Baron,* Mark A. Perrella,* Laura Santambrogio,† Eric S. Silverman,* and Richard J. Riese2* Cathepsin S is a cysteine protease with potent endoproteolytic activity and a broad pH profile. Cathepsin S activity is essential for complete processing of the MHC class II-associated invariant chain within B cells and dendritic cells, and may also be important in extracellular matrix degradation in atherosclerosis and emphysema. Unique among cysteine proteases, cathepsin S activity is up-regulated by IFN-␥. Given its importance, we sought to elucidate the pathway by which IFN-␥ increases cathepsin S expression. Our data demonstrate that the cathepsin S promoter contains an IFN-stimulated response element (ISRE) that is critical for IFN-␥- Downloaded from induced gene transcription in a cell line derived from type II alveolar epithelial (A549) cells. IFN response factor (IRF)-2 derived from A549 nuclear extracts associates with the ISRE oligonucleotide in gel shift assays, but is quickly replaced by IRF-1 following stimulation with IFN-␥. The time course of IRF-1/ISRE complex formation correlates with increased levels of IRF-1 protein and cathepsin S mRNA. Overexpression of IRF-1, but not IRF-2, markedly augments cathepsin S promoter activity in A549 cells. Furthermore, overexpression of IRF-1 increases endogenous cathepsin S mRNA levels in 293T epithelial cells. Finally, freshly isolated bone marrow cells from IRF-1؊/؊ mice fail to up-regulate cathepsin S activity in response to IFN-␥. Thus, IRF-1 is the critical transcriptional mediator of http://www.jimmunol.org/ IFN-␥-dependent cathepsin S activation. These data elucidate a new pathway by which IRF-1 may affect MHC class II processing and presentation. The Journal of Immunology, 2002, 168: 4488–4494. athepsin S is a member of the papain-like family of cys- liberation of the Ii remnant from the class II-binding groove and teine proteases that reside within the endosomal compart- class II-peptide complex formation. The importance of cathepsin S ments and mediate proteolysis of endocytosed proteins in regulating MHC class II-restricted Ag processing and presen- C Ϫ/Ϫ and polypeptides (1–3). Among this family, cathepsin S has unique tation is well illustrated by the phenotype of cathepsin S mice features pertinent to its biological activity. First, it exhibits a broad (6, 7). Inhibition of cathepsin S activity in B cells and dendritic pH profile and maintains a significant endoproteolytic activity at cells results in accumulation of class II-Ii complexes, attenuation by guest on September 26, 2021 neutral pH, suggesting that this enzyme may have important ex- of class II-peptide complex formation, and inability of these cells tracellular biological activity (1). Second, its constitutive expres- to present certain antigenic determinants. Dendritic cells derived sion is restricted primarily to bone marrow-derived APCs (4). from cathepsin SϪ/Ϫ mice have a marked defect of MHC class II Third, its mRNA expression, protein, and activity are up-regulated endosomal trafficking (9, 10). Finally, in human dendritic cells, by IFN-␥ (3) in several cell types (1, 2). proinflammatory cytokines give rise to a surge of MHC class II- Cathepsin S plays an essential role in regulation of MHC class peptide complex formation that is dependent on cathepsin S ac- II maturation and trafficking within APCs by mediating processing tivity (11). Thus, regulation of cathepsin S expression and activity 3 of class II-associated invariant chain (Ii) and exogenous Ags (4– has important consequences in control of MHC class II function 8). In B cells and dendritic cells, cathepsin S uniquely mediates the and subsequent CD4ϩ T cell stimulation. final cleavage of Ii to generate class II-associated invariant chain IFN-␥ is a potent regulator of cathepsin S expression in vascular peptide (CLIP). This cleavage is necessary to permit subsequent smooth muscle cells and in the lung parenchyma (12). Arterial smooth muscle cells stimulated with IFN-␥ exhibit enhanced ca- *Department of Medicine, Brigham and Women’s Hospital and Harvard Medical thepsin S activity and secretion, with a concomitant increase in School, Boston, MA 02115; and †Department of Cancer Immunology and AIDS, supernatant elastolytic activity, which is blocked with a selective Dana-Farber Cancer Institute, Boston, MA 02115 cathepsin S inhibitor (13). Furthermore, cathepsin S activity is in- Received for publication October 23, 2001. Accepted for publication February creased in walls of atheromas and aneurysms as compared with 28, 2002. normal arteries, suggesting that this enzyme may be playing a role The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance in breakdown of the extracellular matrix in atherosclerosis (14). with 18 U.S.C. Section 1734 solely to indicate this fact. Within the lung, transgenic mice generated to overexpress IFN-␥ 1 This work was supported by National Institutes of Health Grants 1UO1 HL65899 exhibit a chronic inflammatory cell infiltrate, increased cysteine and KO8 AI01555 (to R.J.R.). K.S.v.G. is the recipient of a grant from the Deutsche protease activity, and emphysematous-like changes in lung pathol- Forschungsgemeinschaft (STO 420/1-1). ogy and physiology (15, 16). Analysis of bronchoalveolar lavage 2 Address correspondence and reprint requests to Dr. Richard J. Riese, Division of Pulmonary and Critical Care Medicine, Brigham and Women’s Hospital, Tower 4B, fluid of these transgenic mice shows that cathepsin S retains the 75 Francis Street, Boston, MA 02115. E-mail address: [email protected] most activity of the cysteine proteases in the extracellular envi- 3 Abbreviations used in this paper: Ii, invariant chain; CIITA, class II transactivator; ronment, consistent with its broad pH profile. Thus, IFN-␥-induced CLIP, class II-associated invariant chain peptide; 125I-JPM565, 125I-labeled JPM565; activation of cathepsin S has important physiological relevance in ICSBP, IFN consensus sequence-binding protein; IRF, IFN regulatory factor; ISGF3␥, IFN-stimulated gene factor 3␥; ISRE, IFN-stimulated response element; several disease processes, and elucidation of this pathway may ␤-gal, ␤-galactosidase. have direct clinical significance. Copyright © 2002 by The American Association of Immunologists 0022-1767/02/$02.00 The Journal of Immunology 4489 IFN-␥ signaling involves ligand-induced oligomerization of Active site labeling of cysteine proteases and IFN-␥ receptor subunits, leading to phosphorylation and activation immunoprecipitation of Janus kinase 1 and 2. This in turn leads to activation of the Cells were lysed in 50 ␮l 1% Triton X-100, 50 mM sodium acetate (pH dormant Stat1 molecule. Stat1 homodimers translocate to the nu- 4.2), and 1 mM EDTA on ice for 30 min. Postnuclear extracts were nor- cleus, where they direct transcription of specific target genes (17), malized to total protein content and incubated with 125I-labeled JPM565 including such secondary transcriptional activators as the family of (125I-JPM565; gift of H. Ploegh, Boston, MA) for 1 h at 37°C. Labeled IFN response factors (IRF), class II transactivator (CIITA) pro- lysates were then either subjected to SDS-PAGE right away (bone marrow ␤ cells), or immunoprecipitation, as described below (A549 cells). moters III and IV, and CCAAT/enhancing-binding protein- (18– Immunoprecipitation was performed, as described previously (21). Fol- 20). These transcriptional regulators are intermediaries in a com- lowing active site labeling of A549 cell lysates with 125I-JPM, SDS was plicated network that produces alterations in gene expression and added to bring the final concentration to 1%. Samples were boiled for 3 cellular function. min, neutralized, and diluted 10-fold with 0.5% Nonidet P-40, 50 mM Tris (pH 7.4), and 5 mM MgCl2. For the immunoprecipitation, samples were This study seeks to identify the molecular pathway by which precleared initially with 5 ␮l normal rabbit serum (Sigma-Aldrich) and 100 IFN-␥ induces up-regulation of the cathepsin S gene. We have ␮l protein A-agarose (Santa Cruz Biotechnology, Santa Cruz, CA), fol- identified a functional IFN-stimulated response element (ISRE) in lowed by a second preclear with 100 ␮l protein A-agarose alone. Immu- the cathepsin S promoter Ϫ100 bp from the transcriptional start noprecipitation was performed by incubating each sample with 10 ␮l anti- ␮ site. We show that, in human bronchial epithelial A549 cells, cathepsin S serum (gift of H.
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