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

ABSTRACT: 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 sub-types within a population of P.ginigivalis strains NCTC 11834 and W50 that display differences in their ability to invade oral epithelial cells. The highly-invasive sub-types invade cells at 10-30 fold higher levels than the poorly-invasive subtype and remain highly invasive for approximately 12-16 generations. Analysis of the gingipain activity of these sub-types 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 DeltargpAB bacteria did not contain a hyperinvasive sub-type. Screening of the protease activity of wild-type populations of both strains identified high and low protease sub-types which also showed the corresponding reduction or enhancement of invasive capabilities. Microarray analysis of these bistable populations revealed a putative signature set of genes that include oxidative stress resistance and iron transport genes that might be key to invasion of or survival within epithelial cells.

ORGANISM(S): Porphyromonas gingivalis

PROVIDER: GSE23021 | GEO | 2010/08/31

SECONDARY ACCESSION(S): PRJNA131489

REPOSITORIES: GEO

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

Project description: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 sub-types within a population of P.ginigivalis strains NCTC 11834 and W50 that display differences in their ability to invade oral epithelial cells. The highly-invasive sub-types invade cells at 10-30 fold higher levels than the poorly-invasive subtype and remain highly invasive for approximately 12-16 generations. Analysis of the gingipain activity of these sub-types 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 DeltargpAB bacteria did not contain a hyperinvasive sub-type. Screening of the protease activity of wild-type populations of both strains identified high and low protease sub-types which also showed the corresponding reduction or enhancement of invasive capabilities. Microarray analysis of these bistable populations revealed a putative signature set of genes that include oxidative stress resistance and iron transport genes that might be key to invasion of or survival within epithelial cells. Porphyromonas gingivalis comparison of Invasive V Non-invasive sub-types of NCTC11834 and W50 (samples 1-12 AND High V low protease subtypes (samples 13-24) Samples 1-3 were compared top 4-6 and 7-9 V 10-12 for invasive experiments and similarly for protease strain experiments

2010-08-31 | E-GEOD-23021 | biostudies-arrayexpress

Polyphenols from Myrothamnus flabellifolia Welw. inhibit in vitro adhesion of Porphyromonas gingivalis and exert anti-inflammatory cytoprotective effects in KB cells

Project description:Investigation of whole genome gene expression level changes in Porphyromonas gingivalis ATCC 33277 treated with an anti-adhesive extract from Myrothamnus flabellifolia compared to the untreated strain. Aim: Identification of anti-adhesive plant extracts against cell surface binding of Porphyromonas gingivalis. Materials and Methods: Polyphenol-enriched extract from Myrothamnus flabellifolia (MF) traditionally used for periodontitis was tested for inhibition of P. gingivalis adhesion to KB cells by FACS, for influence on gingipain activity, hemagglutination and by microarray analysis for effects on the bacterial transcriptome. P. gingivalis-induced inflammation parameters were monitored by RT-PCR. Results: MF (100 µg/ml) reduced P. gingivalis adhesion/invasion about 50% by interacting with fimbriae and bacterial OMPs. Microarray analysis of MF-treated bacteria indicated up-regulation of genes involved in cell adhesion. As confirmed by RT-PCR, fimbrillin- and Arg-gingipain-encoding genes were upregulated by MF. On the protein level, inhibition (70%) of Arg-gingipain activity was observed, while the corresponding Lys-gingipain was hardly influenced. MF also inhibited hemagglutination. While exposure to P. gingivalis resulted in an increased expression of inflammation-related genes in KB cells, pretreatment of KB cells with MF evoked cytoprotective effects concerning IL-1β, IL-6, IL-8 and TNFα gene expression as well as IL-6 release rates. Conclusions: While being cytoprotective, MF exerts strong anti-adhesive effects against P. gingivalis. Thus, MF may be useful for the prevention of P. gingivalis-associated periodontal diseases. The chip study used total RNA recovered from two separate MF-treated and two separate untreated Porphyromonas gingivalis ATCC 33277 cultures. Each chip measured the expression level of 1,842 genes from P. gingivalis ATCC 33277 with thirteen 60-mer probes per gene, with three-fold technical redundancy.

2010-09-02 | E-GEOD-23939 | biostudies-arrayexpress

Polyphenols from Myrothamnus flabellifolia Welw. inhibit in vitro adhesion of Porphyromonas gingivalis and exert anti-inflammatory cytoprotective effects in KB cells

2010-09-02 | GSE23939 | GEO

HCT116 gene expression analysis upon CAF stimulation

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2017-08-31 | GSE78947 | GEO

Transcription profiling of human intratumoral samples from central and invasive regions of seven widely invasive PTCs and normal thyroid tissue

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2008-06-14 | E-GEOD-6004 | biostudies-arrayexpress

Transcriptional profiling and comparative genomic analysis of an invasive and non-invasive strain of Candida albicans

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2008-03-11 | GSE10689 | GEO

Transcriptional profiling and comparative genomic analysis of an invasive and non-invasive strain of Candida albicans

2010-05-26 | E-GEOD-10689 | biostudies-arrayexpress

Expression data from CL1-0 and CL1-5 lung cancer cell line.

Project description:A lung cancer cell model of invasive transformation was developed to select progressively invasive cell populations from a parental cell line of human lung adenocarcinoma, CL1. Five progressive sub-clones namely, CL1-1, CL1-2, CL1-3 CL1-4, and CL1-5 were selected using transwell and displayed increasing invasion potential (Chu et al, 1997). Here, we used microarrays to analyze and compare gene expression profiles between CL1-0 and CL1-5 for the identification of invasion/metastasis associated gene signatures.

2012-12-11 | GSE42407 | GEO

Integrative analysis of mesenchymal stromal cell conversion by breast cancer secretomes in blank slate 3D microenvironments

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2019-05-27 | PXD010467 | Pride

TGFβ1 promotes collective invasion of oncogenically transformed intestinal organoids by inducing partial epithelial-mesenchymal transition independently of Snail1 and Zeb1

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