A Sialidase‐Deficient Porphyromonas gingivalis Mutant Strain Induces Less Interleukin‐1&bgr; and Tumor Necrosis Factor‐&agr; in Epi4 Cells Than W83 Strain Through Regulation of c‐Jun N‐Terminal Kinase Pathway

BACKGROUND Porphyromonas gingivalis is one of the major periodontal pathogens. In a previous study, a mouse abscess model showed that sialidase deficiency of P. gingivalis weakened its virulence, but the mechanism behind this observation remains unknown. METHODS A sialidase-deficient mutant strain (△PG0352) and a complemented strain (com△PG0352) were constructed. Epi4 cells were stimulated by wild-type strain P. gingivalis W83, △PG0352, or com△PG0352. Real-time polymerase chain reaction was carried out to detect expression of virulent genes in P. gingivalis and interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α in epi4 cells. Activities of sialidase, gingipains, and lipopolysaccharide (LPS) were compared among the different P. gingivalis strains. Levels of IL-1β and TNF-α in the epi4 cells supernatant were detected by enzyme-linked immunosorbent assay and levels of p38, extracellular signal-regulated kinase, c-Jun N-terminal kinase (JNK), and phospho-c-Jun were detected by western blotting. RESULTS Compared with P. gingivalis W83 and com△PG0352, activities of Kgp and Rgp gingipains and amount of LPS decreased in △PG0352, whereas there were no differences in LPS activity among these three strains. Level of phospho-JNK was lower in epi4 cells stimulated by △PG0352. △pG0352 induced less IL-1β and TNF-α and more IL-8 in epi4 cells; differences in IL-1β and TNF-α could not be detected after JNK blocking. CONCLUSION A sialidase-deficient P. gingivalis mutant strain induces less IL-1β and TNF-α in epi4 cells than W83 strain through regulation of JNK pathway.

[1]  K. Tabeta,et al.  Resveratrol suppresses the inflammatory responses of human gingival epithelial cells in a SIRT1 independent manner. , 2015, Journal of periodontal research.

[2]  Shu-Hui Zhang,et al.  Panax Notoginseng flower saponins (PNFS) inhibit LPS-stimulated NO overproduction and iNOS gene overexpression via the suppression of TLR4-mediated MAPK/NF-kappa B signaling pathways in RAW264.7 macrophages , 2015, Chinese Medicine.

[3]  B. Bueno-Silva,et al.  Mechanisms Involved in the Association between Periodontitis and Complications in Pregnancy , 2015, Front. Public Health.

[4]  D. Deng,et al.  The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro , 2015, Journal of oral microbiology.

[5]  Zhongchen Song,et al.  miRNA-146 negatively regulates the production of pro-inflammatory cytokines via NF-κB signalling in human gingival fibroblasts , 2014, Journal of Inflammation.

[6]  T. Thurnheer,et al.  Role of Porphyromonas gingivalis gingipains in multi-species biofilm formation , 2014, BMC Microbiology.

[7]  H. Fletcher,et al.  Involvement of PG2212 Zinc Finger Protein in the Regulation of Oxidative Stress Resistance in Porphyromonas gingivalis W83 , 2014, Journal of bacteriology.

[8]  T. Fujita,et al.  Amphotericin B down-regulates Aggregatibacter actinomycetemcomitans-induced production of IL-8 and IL-6 in human gingival epithelial cells. , 2014, Cellular immunology.

[9]  S. Wallet,et al.  Deletion of Lipoprotein PG0717 in Porphyromonas gingivalis W83 Reduces Gingipain Activity and Alters Trafficking in and Response by Host Cells , 2013, PloS one.

[10]  Chunhao Li,et al.  A surface‐exposed neuraminidase affects complement resistance and virulence of the oral spirochaete Treponema denticola , 2013, Molecular microbiology.

[11]  M. Ogrendik Rheumatoid arthritis is an autoimmune disease caused by periodontal pathogens , 2013, International journal of general medicine.

[12]  R. Darveau,et al.  Nuclear factor-κB and p38 mitogen-activated protein kinase signaling pathways are critically involved in Porphyromonas gingivalis lipopolysaccharide induction of lipopolysaccharide-binding protein expression in human oral keratinocytes. , 2013, Molecular oral microbiology.

[13]  Cun-Yu Wang,et al.  Baicalin Downregulates Porphyromonas gingivalis Lipopolysaccharide-Upregulated IL-6 and IL-8 Expression in Human Oral Keratinocytes by Negative Regulation of TLR Signaling , 2012, PloS one.

[14]  N. Bostanci,et al.  Doxycycline inhibits TREM-1 induction by Porphyromonas gingivalis. , 2012, FEMS immunology and medical microbiology.

[15]  N. Bostanci,et al.  Porphyromonas gingivalis: an invasive and evasive opportunistic oral pathogen. , 2012, FEMS microbiology letters.

[16]  J. Vaqué,et al.  Interleukin-21 expression and its association with proinflammatory cytokines in untreated chronic periodontitis patients. , 2012, Journal of periodontology.

[17]  Jun Liu,et al.  Abrogation of Neuraminidase Reduces Biofilm Formation, Capsule Biosynthesis, and Virulence of Porphyromonas gingivalis , 2011, Infection and Immunity.

[18]  K. Tabeta,et al.  Effect of interleukin-17 on the expression of chemokines in gingival epithelial cells. , 2011, European journal of oral sciences.

[19]  H. Fletcher,et al.  Sialidase and Sialoglycoproteases Can Modulate Virulence in Porphyromonas gingivalis , 2011, Infection and Immunity.

[20]  Andrew E. Parker,et al.  Targeting Toll-like receptors: emerging therapeutics? , 2010, Nature Reviews Drug Discovery.

[21]  C. Douglas,et al.  A Novel Sialic Acid Utilization and Uptake System in the Periodontal Pathogen Tannerella forsythia , 2010, Journal of bacteriology.

[22]  D. Kinane,et al.  The host cytokine response to Porphyromonas gingivalis is modified by gingipains. , 2009, Oral microbiology and immunology.

[23]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[24]  Y. Abiko,et al.  Role of the hemin-binding protein 35 (HBP35) of Porphyromonas gingivalis in coaggregation. , 2008, Microbial pathogenesis.

[25]  Maki Maeda,et al.  Role for Gingipains in Porphyromonas gingivalis Traffic to Phagolysosomes and Survival in Human Aortic Endothelial Cells , 2007, Infection and Immunity.

[26]  H. Fletcher,et al.  VimA is part of the maturation pathway for the major gingipains of Porphyromonas gingivalis W83. , 2006, Microbiology.

[27]  Liang Tong,et al.  Bacterial neuraminidase facilitates mucosal infection by participating in biofilm production. , 2006, The Journal of clinical investigation.

[28]  R. Lamont,et al.  LuxS Involvement in the Regulation of Genes Coding for Hemin and Iron Acquisition Systems in Porphyromonas gingivalis , 2006, Infection and Immunity.

[29]  J. P. Long,et al.  Immunization with Recombinant Streptococcus pneumoniae Neuraminidase NanA Protects Chinchillas against Nasopharyngeal Colonization , 2005, Infection and Immunity.

[30]  M. Quirynen,et al.  Differential induction of human beta-defensin expression by periodontal commensals and pathogens in periodontal pocket epithelial cells. , 2005, Journal of periodontology.

[31]  S. Holt,et al.  Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia: the "red complex", a prototype polybacterial pathogenic consortium in periodontitis. , 2005, Periodontology 2000.

[32]  S. Haake,et al.  Differential regulation of cytokine genes in gingival epithelial cells challenged by Fusobacterium nucleatum and Porphyromonas gingivalis. , 2004, Microbial pathogenesis.

[33]  S. Way,et al.  Porphyromonas gingivalis Lipopolysaccharide Contains Multiple Lipid A Species That Functionally Interact with Both Toll-Like Receptors 2 and 4 , 2004, Infection and Immunity.

[34]  G. Garlet,et al.  Matrix metalloproteinases, their physiological inhibitors and osteoclast factors are differentially regulated by the cytokine profile in human periodontal disease. , 2004, Journal of clinical periodontology.

[35]  Y. Shimabukuro,et al.  Human gingival epithelial cells produce chemotactic factors interleukin-8 and monocyte chemoattractant protein-1 after stimulation with Porphyromonas gingivalis via toll-like receptor 2. , 2004, Journal of periodontology.

[36]  P. Veith,et al.  Porphyromonas gingivalis gingipains: the molecular teeth of a microbial vampire. , 2003, Current protein & peptide science.

[37]  R. Genco,et al.  Periodontal disease and cardiovascular disease: epidemiology and possible mechanisms. , 2002, Journal of the American Dental Association.

[38]  Y. Shimabukuro,et al.  Activation of Adenosine-receptor-enhanced iNOS mRNA Expression by Gingival Epithelial Cells , 2002, Journal of dental research.

[39]  K. Watanabe,et al.  Association of Mitogen-Activated Protein Kinase Pathways with Gingival Epithelial Cell Responses to Porphyromonas gingivalis Infection , 2001, Infection and Immunity.

[40]  D. Beighton,et al.  Isolation and characterisation of sialidase from a strain of Streptococcus oralis. , 2000, Journal of medical microbiology.

[41]  E. Hounsell,et al.  Variable Carbohydrate Modifications to the Catalytic Chains of the RgpA and RgpB Proteases of Porphyromonas gingivalis W50 , 1999, Infection and Immunity.

[42]  Yixin Shi,et al.  Genetic Analyses of Proteolysis, Hemoglobin Binding, and Hemagglutination of Porphyromonas gingivalis , 1999, The Journal of Biological Chemistry.

[43]  W. Knowler,et al.  Severe Periodontitis and Risk for Poor Glycemic Control in Patients with Non-Insulin-Dependent Diabetes Mellitus. , 1996, Journal of periodontology.

[44]  M. Curtis,et al.  Development of a simple chemically defined medium for Porphyromonas gingivalis: requirement for α-ketoglutarate , 1996 .

[45]  S. Hillier,et al.  Sialidase (neuraminidase) activity among gram-negative anaerobic and capnophilic bacteria , 1990, Journal of clinical microbiology.

[46]  B. Moncla,et al.  Detection of sialidase (neuraminidase) activity in Actinomyces species by using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid in a filter paper spot test , 1989, Journal of clinical microbiology.

[47]  S. Gorr Antimicrobial peptides in periodontal innate defense. , 2012, Frontiers of oral biology.

[48]  M. Hackett,et al.  Rapid isolation method for lipopolysaccharide and lipid A from gram-negative bacteria. , 2000, The Analyst.

[49]  M. Curtis,et al.  Development of a simple chemically defined medium for Porphyromonas gingivalis: requirement for alpha-ketoglutarate. , 1996, FEMS microbiology letters.

[50]  S. Socransky,et al.  The bacterial etiology of destructive periodontal disease: current concepts. , 1992, Journal of periodontology.