<i>Porphyromonas gingivalis</i> survives within KB cells and modulates inflammatory response

Eick, Sigrun; Reissmann, A.; Rödel, Jürgen; Schmidt, Karl‐Hermann; Pfister, W.

doi:10.1111/j.1399-302x.2006.00282.x

Cited by 33 publications

(28 citation statements)

References 42 publications

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“…1b,. This is in agreement with other published data that used KB cells (Dorn et al, 2000;Eick et al, 2006;Houalet-Jeanne et al, 2001), and similar levels of invasion have been reported using normal oral keratinocytes (Lamont et al, 1995). When the invasion levels of the recovered bacteria were analysed after a second round of invasion with fresh cells we observed an increase in the percentage of bacteria able to invade the OSCC cells for both strains of over threefold (Fig.…”

Section: Resultssupporting

confidence: 83%

“…The mechanism of epithelial cell invasion by P. gingivalis has not been fully elucidated but has been suggested to depend upon the interaction of bacterial fimbriae with cell-associated fibronectin and the fibronectin-binding integrin a 5 b 1 (Tsuda et al, 2008;Yilmaz et al, 2002;Yilmaz, 2008). However, there have also been reports that strains with identical fimbriation characteristics vary in their invasive ability (Dorn et al, 2000;Duncan et al, 1993;Hamada et al, 1994;Eick et al, 2006), indicating uncertainty about the mechanisms employed by all strains of P. gingivalis and that more than one mechanism may be operating.…”

Section: Introductionmentioning

confidence: 99%

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Identification of bistable populations of Porphyromonas gingivalis that differ in epithelial cell invasion

et al. 2010

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Bistable populations of bacteria give rise to two or more subtypes that exhibit different phenotypes. We have explored whether the periodontal pathogen Porphyromonas gingivalis exhibits bistable invasive phenotypes. Using a modified cell invasion assay, we show for the first time that there are two distinct subtypes within a population of P. gingivalis strains NCTC 11834 and W50 that display differences in their ability to invade oral epithelial cells. The highly invasive subtype invades cells at 10-30-fold higher levels than the poorly invasive subtype and remains highly invasive for approximately 12-16 generations. Analysis of the gingipain activity of these subtypes revealed that the highly invasive type had reduced cell-associated arginine-specific protease activity. The role of Arg-gingipain activity in invasion was verified by enhancement of invasion by rgpAB mutations and by inclusion of an Arg-gingipain inhibitor in invasion assays using wild-type bacteria. In addition, a population of DrgpAB bacteria did not contain a hyperinvasive subtype. Screening of the protease activity of wild-type populations of both strains identified high and low protease subtypes which also showed a corresponding reduction or enhancement, respectively, of invasive capabilities. Microarray analysis of these bistable populations revealed a putative signature set of genes that includes oxidative stress resistance and iron transport genes, and which might be critical to invasion of or survival within epithelial cells.

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Section: Resultssupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Identification of bistable populations of Porphyromonas gingivalis that differ in epithelial cell invasion

et al. 2010

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“…Virulence of P. gingivalis against oral epithelial cells has often been characterized in KB cells (Eick et al, 2006;Madianos et al, 1996;Pathirana et al, 2007b). Although KB cells were originally derived from an oral epidermal carcinoma, further analysis demonstrated HeLa markers in the chromosomes.…”

Section: Discussionmentioning

confidence: 99%

Cleavage of protease-activated receptors on an immortalized oral epithelial cell line by Porphyromonas gingivalis gingipains

et al. 2009

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Porphyromonas gingivalis activates protease-activated receptors (PARs) on oral keratinocytes, resulting in downstream signalling for an innate immune response. Activation depends on P. gingivalis gingipains, but could be confounded by lipopolysaccharide signalling through Toll-like receptors. We therefore hypothesized that P. gingivalis cleaves oral keratinocyte PARs in an Arg-(Rgp) or Lys-(Kgp) gingipain-specific manner to upregulate pro-inflammatory cytokines. Immortalized human oral keratinocytes (TERT-2) were incubated with wild-type P. gingivalis (ATCC 33277) or strains from a panel of isogenic gingipain deletion mutants: Kgp-deficient (KDP 129); Rgp-deficient (KDP 133); or Kgp-and Rgp-deficient (KDP 136). After incubation with P. gingivalis, keratinocytes were probed with specific antibodies against the N-terminus of PAR-1 and PAR-2. Using flow cytometry and immunofluorescence, receptor cleavage was marked by loss of specific antibody binding to the respective PARs. TERT-2 cells constitutively expressed high levels of PAR-1 and PAR-2, and lower levels of PAR-3. P. gingivalis ATCC 33277 cleaved PAR-1 and PAR-2 in a dose-dependent manner, while the receptors were unaffected by the protease-negative double mutant (KDP 136) at all m.o.i. tested. The single Kgp-negative mutant preferentially cleaved PAR-1, whereas the Rgp-negative mutant cleaved PAR-2. Wild-type or Kgp-negative mutant cleavage of PAR-1 upregulated expression of IL-1a, IL-1b, IL-6 and TNF-a; the Rgp-negative mutant did not modulate these cytokines. Selective cleavage of PAR-1 on oral epithelial cells by P. gingivalis Rgp therefore upregulates expression of pro-inflammatory cytokines.

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“…To confirm any changes in essential P. gingivalis activity under varying non-optimal heme concentrations, Rgp activity was determined following a previously published work [18]. Briefly, 0.5 mM N-a-benzoyl-DL-arginine-p-nitroanilide (BApNA; Sigma) and enzyme activity buffer (0.2 M TriseHCl, 0.1 M NaCl, pH 7.5) were mixed with the disrupted bacterial cells and, subsequently, incubated at 37 C. Bacterial suspension was measured using a spectrometer (Abs 405 nm).…”

mentioning

confidence: 99%

Similar physiological effects in Porphyromonas gingivalis ATCC 33277 under hemin-excess and hemin-limited concentrations are putatively associated to different hydrogen peroxide function

et al. 2014

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Porphyromonas gingivalis survives within KB cells and modulates inflammatory response

Abstract: The results indicate survival of P. gingivalis within epithelial cells, possibly in a non-cultivable stage. Invasion into cells modulates the virulence properties of P. gingivalis as well as the inflammatory response of the cells.

Cited by 33 publications

References 42 publications

Identification of bistable populations of Porphyromonas gingivalis that differ in epithelial cell invasion

Identification of bistable populations of Porphyromonas gingivalis that differ in epithelial cell invasion

Cleavage of protease-activated receptors on an immortalized oral epithelial cell line by Porphyromonas gingivalis gingipains

Similar physiological effects in Porphyromonas gingivalis ATCC 33277 under hemin-excess and hemin-limited concentrations are putatively associated to different hydrogen peroxide function

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