Integrin α5β1-fimbriae binding and actin rearrangement are essential for Porphyromonas gingivalis invasion of osteoblasts and subsequent activation of the JNK pathway

BMC Microbiology

Gena Tribble¹  Todd Rigney¹  Jun Ju²  Wenjian Zhang² 

[1] Department of Periodontics and Dental Hygiene, University of Texas School of Dentistry at Houston, 7500 Cambridge Street, Houston, TX, 77054, USA;Department of Diagnostic and Biomedical Sciences, University of Texas School of Dentistry at Houston, 7500 Cambridge Street, Suite 5366, Houston, TX, 77054, USA

关键词: Apoptosis;    Signaling;    Cytoskeleton;    Integrins;    Porphyromonas gingivalis;    Osteoblasts;   

Others  :  1144609
DOI  :  10.1186/1471-2180-13-5

 received in 2012-10-23, accepted in 2013-01-08,  发布年份 2013

【 摘 要 】

Background

Chronic periodontitis is an infectious disease of the periodontium, which includes the gingival epithelium, periodontal ligament and alveolar bone. The signature clinical feature of periodontitis is resorption of alveolar bone and subsequent tooth loss. The Gram-negative oral anaerobe, Porphyromonas gingivalis, is strongly associated with periodontitis, and it has been shown previously that P. gingivalis is capable of invading osteoblasts in a dose- and time-dependent manner resulting in inhibition of osteoblast differentiation and mineralization in vitro. It is not yet clear which receptors and cytoskeletal components mediate the invasive process, nor how the signaling pathways and viability of osteoblasts are affected by bacterial internalization. This study aimed to investigate these issues using an in vitro model system involving the inoculation of P. gingivalis ATCC 33277 into primary osteoblast cultures.

Results

It was found that binding between P. gingivalis fimbriae and integrin α5β1 on osteoblasts, and subsequent peripheral condensation of actin, are essential for entry of P. gingivalis into osteoblasts. The JNK pathway was activated in invaded osteoblasts, and apoptosis was induced by repeated infections.

Conclusions

These observations indicate that P. gingivalis manipulates osteoblast function to promote its initial intracellular persistence by prolonging the host cell life span prior to its intercellular dissemination via host cell lysis. The identification of molecules critical to the interaction between P. gingivalis and osteoblasts will facilitate the development of new therapeutic strategies for the prevention of periodontal bone loss.

【 授权许可】

   
2013 Zhang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Lamont RJ, Jenkinson HF: Life below the gum line: pathogenic mechanisms of Porphyromonas gingivalis. Microbiol Mol Biol Rev 1998, 62(4):1244-1263.
• [2]Amornchat C, Rassameemasmaung S, Sripairojthikoon W, Swasdison S: Invasion of Porphyromonas gingivalis into human gingival fibroblasts in vitro. J Int Acad Periodontol 2003, 5(4):98-105.
• [3]Frank RM, Voegel JC: Bacterial bone resorption in advanced cases of human periodontitis. J Periodontal Res 1978, 13(3):251-261.
• [4]Lamont RJ, Chan A, Belton CM, Izutsu KT, Vasel D, Weinberg A: Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun 1995, 63(10):3878-3885.
• [5]Zhang W, Ju J, Rigney T, Tribble GD: Fimbriae of Porphyromonas gingivalis are important for initial invasion of osteoblasts, but not for inhibition of their differentiation and mineralization. J Periodontol 2011, 82(6):909-916.
• [6]Zhang W, Swearingen EB, Ju J, Rigney T, Tribble GD: Porphyromonas gingivalis invades osteoblasts and inhibits bone formation. Microbes Infect 2010, 12(11):838-845.
• [7]Ozeri V, Rosenshine I, Ben-Ze’Ev A, Bokoch GM, Jou TS, Hanski E: De novo formation of focal complex-like structures in host cells by invading Streptococci. Mol Microbiol 2001, 41(3):561-573.
• [8]Agerer F, Lux S, Michel A, Rohde M, Ohlsen K, Hauck CR: Cellular invasion by Staphylococcus aureus reveals a functional link between focal adhesion kinase and cortactin in integrin-mediated internalisation. J Cell Sci 2005, 118(Pt 10):2189-2200.
• [9]Plancon L, Du Merle L, Le Friec S, Gounon P, Jouve M, Guignot J, Servin A, Le Bouguenec C: Recognition of the cellular beta1-chain integrin by the bacterial AfaD invasin is implicated in the internalization of afa-expressing pathogenic Escherichia coli strains. Cell Microbiol 2003, 5(10):681-693.
• [10]Amano A: Molecular interaction of Porphyromonas gingivalis with host cells: implication for the microbial pathogenesis of periodontal disease. J Periodontol 2003, 74(1):90-96.
• [11]Tsuda K, Furuta N, Inaba H, Kawai S, Hanada K, Yoshimori T, Amano A: Functional analysis of alpha5beta1 integrin and lipid rafts in invasion of epithelial cells by Porphyromonas gingivalis using fluorescent beads coated with bacterial membrane vesicles. Cell Struct Funct 2008, 33(1):123-132.
• [12]Yilmaz O, Watanabe K, Lamont RJ: Involvement of integrins in fimbriae-mediated binding and invasion by Porphyromonas gingivalis. Cell Microbiol 2002, 4(5):305-314.
• [13]Schoenwaelder SM, Burridge K: Bidirectional signaling between the cytoskeleton and integrins. Curr Opin Cell Biol 1999, 11(2):274-286.
• [14]Young VB, Falkow S, Schoolnik GK: The invasin protein of Yersinia enterocolitica: internalization of invasin-bearing bacteria by eukaryotic cells is associated with reorganization of the cytoskeleton. J Cell Biol 1992, 116(1):197-207.
• [15]Yilmaz O, Young PA, Lamont RJ, Kenny GE: Gingival epithelial cell signalling and cytoskeletal responses to Porphyromonas gingivalis invasion. Microbiology 2003, 149(Pt 9):2417-2426.
• [16]Maniotis AJ, Chen CS, Ingber DE: Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure. Proc Natl Acad Sci U S A 1997, 94(3):849-854.
• [17]Rosengart MR, Arbabi S, Bauer GJ, Garcia I, Jelacic S, Maier RV: The actin cytoskeleton: an essential component for enhanced TNFalpha production by adherent monocytes. Shock (Augusta, Ga) 2002, 17(2):109-113.
• [18]Watanabe K, Yilmaz O, Nakhjiri SF, Belton CM, Lamont RJ: Association of mitogen-activated protein kinase pathways with gingival epithelial cell responses to Porphyromonas gingivalis infection. Infect Immun 2001, 69(11):6731-6737.
• [19]Mao S, Park Y, Hasegawa Y, Tribble GD, James CE, Handfield M, Stavropoulos MF, Yilmaz O, Lamont RJ: Intrinsic apoptotic pathways of gingival epithelial cells modulated by Porphyromonas gingivalis. Cell Microbiol 2007, 9(8):1997-2007.
• [20]Nakhjiri SF, Park Y, Yilmaz O, Chung WO, Watanabe K, El-Sabaeny A, Park K, Lamont RJ: Inhibition of epithelial cell apoptosis by Porphyromonas gingivalis. FEMS Microbiol Lett 2001, 200(2):145-149.
• [21]Urnowey S, Ansai T, Bitko V, Nakayama K, Takehara T, Barik S: Temporal activation of anti- and pro-apoptotic factors in human gingival fibroblasts infected with the periodontal pathogen, Porphyromonas gingivalis: potential role of bacterial proteases in host signalling. BMC Microbiol 2006, 6:26. BioMed Central Full Text
• [22]Yilmaz O, Jungas T, Verbeke P, Ojcius DM: Activation of the phosphatidylinositol 3-kinase/Akt pathway contributes to survival of primary epithelial cells infected with the periodontal pathogen Porphyromonas gingivalis. Infect Immun 2004, 72(7):3743-3751.
• [23]Wong GL, Cohn DV: Target cells in bone for parathormone and calcitonin are different: enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces. Proc Natl Acad Sci U S A 1975, 72(8):3167-3171.
• [24]Elkaim R, Obrecht-Pflumio S, Tenenbaum H: Paxillin phosphorylation and integrin expression in osteoblasts infected by Porphyromonas gingivalis. Arch Oral Biol 2006, 51(9):761-768.
• [25]Waterman-Storer CM: Microtubules and microscopes: how the development of light microscopic imaging technologies has contributed to discoveries about microtubule dynamics in living cells. Mol Biol Cell 1998, 9(12):3263-3271.
• [26]Andrian E, Grenier D, Rouabhia M: Porphyromonas gingivalis-epithelial cell interactions in periodontitis. J Dent Res 2006, 85(5):392-403.
• [27]Robinson MJ, Cobb MH: Mitogen-activated protein kinase pathways. Curr Opin Cell Biol 1997, 9(2):180-186.
• [28]Wang PL, Sato K, Oido M, Fujii T, Kowashi Y, Shinohara M, Ohura K, Tani H, Kuboki Y: Involvement of CD14 on human gingival fibroblasts in Porphyromonas gingivalis lipopolysaccharide-mediated interleukin-6 secretion. Arch Oral Biol 1998, 43(9):687-694.
• [29]Matsuguchi T, Chiba N, Bandow K, Kakimoto K, Masuda A, Ohnishi T: JNK activity is essential for Atf4 expression and late-stage osteoblast differentiation. J Bone Miner Res 2009, 24(3):398-410.
• [30]Mukai T, Otsuka F, Otani H, Yamashita M, Takasugi K, Inagaki K, Yamamura M, Makino H: TNF-alpha inhibits BMP-induced osteoblast differentiation through activating SAPK/JNK signaling. Biochem Biophys Res Commun 2007, 356(4):1004-1010.