Modulation of expression of innate immunity markers CXCL5/ENA-78 and CCL20/MIP3α by protease-activated receptors (PARs) in human gingival epithelial cells
Free accessOtherFirst published online April, 2010
Modulation of expression of innate immunity markers CXCL5/ENA-78 and CCL20/MIP3α by protease-activated receptors (PARs) in human gingival epithelial cells
Protease-activated receptors (PARs) are G-protein-coupled receptors with an active role in host defense. The two most highly expressed members of the PAR family in gingival epithelial cells (GECs) are PAR1 and PAR2. The major virulence factors of periodontal pathogen Porphyromonas gingivalis are its proteases which can activate PAR2. However, little is known about the function of PARs in GECs when they are activated by their endogenous agonist enzymes. The purpose of this study was to characterize how the expression of innate immune markers is modulated when PAR1 and PAR2 are activated by their agonist enzymes, thrombin and trypsin, respectively. Here, we report that activation of PAR1 and PAR2 induces cell proliferation at low concentration. Activation of PAR via proteolytic activity of thrombin and trypsin induces expression of CXCL5/ENA-78 and CCL20/MIP3α in a concentration-dependent manner. Induction of CXCL5 via PAR1 was inhibited in the presence of PAR1 cleavage blocking antibodies and by PAR1 siRNA. The induction of CXCL5 and CCL20 via PAR2 was inhibited by PAR2 siRNA. These findings indicate an active role in innate immune responses by PAR1 and PAR2 in GECs. Modulation of innate immunity by PARs may contribute to co-ordinated and balanced immunosurveillance in GECs.
Sugawara Y., Uehara A., Fujimoto Y. et al. Toll-like receptors, NOD1, and NOD2 in oral epithelial cells. J Dent Res2006; 85: 524-529.
2.
Vu TK, Hung DT, Wheaton VI, Coughlin SRMolecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell1991 ; 64: 1057-1068.
3.
Nakanishi-Matsui M., Zheng YW, Sulciner DJ, Weiss EJ, Ludeman MJ, Coughlin SRPAR3 is a cofactor for PAR4 activation by thrombin. Nature2000 ; 404: 609-613.
4.
Coughlin SRProtease-activated receptors in hemostasis, thrombosis and vascular biology . J Thromb Haemost2005; 3: 1800-1814.
Steinhoff M. , Buddenkotte J., Shpacovitch V. et al. Proteinase-activated receptors: transducers of proteinase-mediated signaling in inflammation and immune response. Endocr Rev2005; 26: 1-43.
MacNaughton WKEpithelial effects of proteinase-activated receptors in the gastrointestinal tract. Mem Inst Oswaldo Cruz2005; 100: 211-215.
9.
Lourbakos A. , Potempa J., Travis J. et al. Arginine-specific protease from Porphyromonas gingivalis activates protease-activated receptors on human oral epithelial cells and induces interleukin-6 secretion. Infect Immun2001; 69: 5121-5130.
Dommisch H. , Chung WO, Rohani MG et al. Protease-activated receptor 2 mediates human beta-defensin 2 and CC chemokine ligand 20 mRNA expression in response to proteases secreted by Porphyromonas gingivalis. Infect Immun2007; 75: 4326-4333.
12.
Uehara A., Sugawara S., Muramoto K., Takada H.Activation of human oral epithelial cells by neutrophil proteinase 3 through protease-activated receptor-2. J Immunol2002 ; 169: 4594-4603.
13.
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 Immunol2003; 170: 5690-5696.
14.
Holzhausen M., Spolidorio LC, Vergnolle N.Proteinase-activated receptor-2 (PAR2) agonist causes periodontitis in rats. J Dent Res2005 ; 84: 154-159.
15.
Fiorucci S. , Mencarelli A., Palazzetti B. et al. Proteinase-activated receptor 2 is an anti-inflammatory signal for colonic lamina propria lymphocytes in a mouse model of colitis. Proc Natl Acad Sci USA2001 ; 98: 13936-13941.
16.
Tatakis DNBlood coagulation factors in periodontal pathophysiology: a review with emphasis on the role of thrombin. Semin Thromb Hemost1992; 18: 28-33.
17.
Imamura T., Banbula A., Pereira PJ, Travis J., Potempa J.Activation of human prothrombin by arginine-specific cysteine proteinases (Gingipains R) from Porphyromonas gingivalis. J Biol Chem2001; 276: 18984-18991.
18.
Eley BM, Cox SWCathepsin B/L-, elastase-, tryptase-, trypsin- and dipeptidyl peptidase IV-like activities in gingival crevicular fluid: a comparison of levels before and after periodontal surgery in chronic periodontitis patients. J Periodontol1992; 63: 412-417.
19.
Chung WO, Dale BAInnate immune response of oral and foreskin keratinocytes: utilization of different signaling pathways by various bacterial species. Infect Immun2004; 72: 352-358.
20.
Gentleman RC , Carey VJ, Bates DM et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol2004; 5: R80.
21.
Wu Z., Irizarry R., Gentleman RC, Murillo FM, Spencer F.A model based background adjustment for oligonucleotide expression arrays . J Am Stat Assoc2004; 99: 909-917.
22.
Al-Shahrour F., Minguez P., Tarraga J. et al. BABELOMICS: a systems biology perspective in the functional annotation of genome-scale experiments. Nucleic Acids Res2006; 34: W472-W476.
23.
Pfaffl MWA new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res2001; 29: e45.
24.
Brodersen DE , Nissen P.The social life of ribosomal proteins . FEBS J2005; 272: 2098-2108.
Hofman P., Butori C., Havet K. et al. Prognostic significance of cortactin levels in head and neck squamous cell carcinoma: comparison with epidermal growth factor receptor status. Br J Cancer2008; 98: 956-964.
27.
Kon S., Ikesue M., Kimura C. et al. Syndecan-4 protects against osteopontin-mediated acute hepatic injury by masking functional domains of osteopontin. J Exp Med2008; 205: 25-33.
28.
O’Brien PJ, Prevost N., Molino M. et al. Thrombin responses in human endothelial cells. Contributions from receptors other than PAR1 include the transactivation of PAR2 by thrombin-cleaved PAR1. J Biol Chem2000; 275: 13502-13509.
29.
Derian CK, Eckardt AJ, Andrade-Gordon P.Differential regulation of human keratinocyte growth and differentiation by a novel family of protease-activated receptors. Cell Growth Differ1997; 8: 743-749.
30.
Algermissen B. , Sitzmann J., Nurnberg W., Laubscher JC, Henz BM, Bauer F.Distribution and potential biologic function of the thrombin receptor PAR-1 on human keratinocytes . Arch Dermatol Res2000; 292: 488-495.
31.
Fager G.Thrombin and proliferation of vascular smooth muscle cells. Circ Res1995; 77: 645-650.
Ahmad R., Knafo L., Xu J., Sindhu ST, Menezes J., Ahmad A.Thrombin induces apoptosis in human tumor cells. Int J Cancer2000; 87: 707-715.
34.
Kaufmann K. , Thiel G.Epidermal growth factor and thrombin induced proliferation of immortalized human keratinocytes is coupled to the synthesis of Egr-1, a zinc finger transcriptional regulator. J Cell Biochem2002; 85: 381-391.
35.
Kim TH, Xiong H., Zhang Z., Ren B.beta-Catenin activates the growth factor endothelin-1 in colon cancer cells . Oncogene2005; 24: 597-604.
36.
Walz A., Burgener R., Car B., Baggiolini M., Kunkel SL, Strieter RMStructure and neutrophil-activating properties of a novel inflammatory peptide (ENA-78) with homology to interleukin 8. J Exp Med1991; 174: 1355-1362.
37.
Rubie C., Frick VO, Wagner M. et al. ELR+ CXC chemokine expression in benign and malignant colorectal conditions. BMC Cancer2008; 8: 178-188.
38.
Park JY, Park KH, Bang S. et al. CXCL5 overexpression is associated with late stage gastric cancer . J Cancer Res Clin Oncol2007; 133: 835-840.
39.
Starner TD, Barker CK, Jia HP, Kang Y., McCray Jr, PBCCL20 is an inducible product of human airway epithelia with innate immune properties. Am J Respir Cell Mol Biol2003 ; 29: 627-633.
40.
Yang D., Chen Q., Hoover DM et al. Many chemokines including CCL20/MIP-3alpha display antimicrobial activity. J Leukoc Biol2003; 74: 448-455.
41.
Yin L., Dale BAActivation of protective responses in oral epithelial cells by Fusobacterium nucleatum and human beta-defensin-2. J Med Microbiol2007; 56: 976-987.
42.
Greenbaum D. , Colangelo C., Williams K., Gerstein M.Comparing protein abundance and mRNA expression levels on a genomic scale. Genome Biol2003 ; 4: 117-124.
43.
McLaughlin JN, Patterson MM, Malik ABProtease-activated receptor-3 (PAR3) regulates PAR1 signaling by receptor dimerization. Proc Natl Acad Sci USA2007; 104: 5662-5667.
44.
Ostrowska E. , Reiser G.The protease-activated receptor-3 (PAR-3) can signal autonomously to induce interleukin-8 release. Cell Mol Life Sci2008; 65: 970-981.
