&lt;em&gt;Porphyromonas gingivalis&lt;/em&gt; as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells

Wunsch, Christopher M; Lewis, Janina P.

doi:10.3791/53408

Cited by 9 publications

(9 citation statements)

References 50 publications

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“…After infection, the cells were washed four times with sterile PBS. The extracellular bacteria were killed by treating the cells with gentamycin (100 μg/mL) for 1 h. After antibiotic treatment, the cells were washed four times, using sterile PBS, and were incubated with 1% saponin for 15 min [ 40 ]. Then, 100 μL of lysate was spread on LB agar and BHI plates and incubated for 24 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Bitter Taste Receptor T2R14 Modulates Gram-Positive Bacterial Internalization and Survival in Gingival Epithelial Cells

Medapati

Bhagirath

Singh

et al. 2021

IJMS

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Bitter-taste receptors (T2Rs) have emerged as key players in host–pathogen interactions and important modulators of oral innate immunity. Previously, we reported that T2R14 is expressed in gingival epithelial cells (GECs) and interacts with competence stimulating peptides (CSPs) secreted by the cariogenic Streptococcus mutans. The underlying mechanisms of the innate immune responses and physiological effects of T2R14 on Gram-positive bacteria are not well characterized. In this study, we examined the role of T2R14 in internalization and growth inhibitory effects on Gram-positive bacteria, namely Staphylococcus aureus and S. mutans. We utilized CRISPR-Cas9 T2R14 knockdown (KD) GECs as the study model to address these key physiological mechanisms. Our data reveal that the internalization of S. aureus is significantly decreased, while the internalization of S. mutans remains unaffected upon knockdown of T2R14 in GECs. Surprisingly, GECs primed with S. mutans CSP-1 resulted in an inhibition of growth for S. aureus, but not for S. mutans. The GECs infected with S. aureus induced T2R14-dependent human β-defensin-2 (hBD-2) secretion; however, S. mutans–infected GECs did not induce hBD-2 secretion, but induced T2R14 dependent IL-8 secretion. Interestingly, our results show that T2R14 KD affects the cytoskeletal reorganization in GECs, thereby inhibiting S. aureus internalization. Our study highlights the distinct mechanisms and a direct role of T2R14 in influencing physiological responses to Gram-positive bacteria in the oral cavity.

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Section: Methodsmentioning

confidence: 99%

Bitter Taste Receptor T2R14 Modulates Gram-Positive Bacterial Internalization and Survival in Gingival Epithelial Cells

Medapati

Bhagirath

Singh

et al. 2021

IJMS

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“…Pg was cultured as described previously 19 . Frozen bacterial Pg (strain W83) stocks were grown anaerobically on Trypticase Soy Agar (TSA) blood agar plates (BAP) with 5% sheep blood at 37°C in a BACTRON‐300 Anaerobic Chamber for 4 to 7 days.…”

Section: Methodsmentioning

confidence: 99%

“…Pg was cultured as described previously. 19 Frozen bacterial Pg (strain W83) stocks were grown anaerobically on Trypticase Soy Agar (TSA) blood agar plates (BAP) with 5% sheep blood at 37 • C in a BACTRON-300 Anaerobic Chamber for 4 to 7 days. Using sterile loops, Pg was inoculated into 3 mL TSA broth supplemented with hemin and menadione, which allowed bacteria to grow until mid-log phase (OD 600 of ≈ 0.8).…”

Section: Pre-culture Of Pgmentioning

confidence: 99%

Porphyromonas gingivalis infection alters Nrf2‐phase II enzymes and nitric oxide in primary human aortic endothelial cells

Sampath

Okoro

Gipson

et al. 2020

Journal of Periodontology

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Background: Periodontal disease (PD) is known to be associated with endothelial dysfunction in patients with coronary artery and/or cardiovascular disease. In our study, we sought to explore the virulence of P. gingivalis (Pg) affecting glycogen synthase kinase 3 beta (GSK-3β)/nuclear factor (erythroid-derived 2)like 2 (Nrf2)/tetrahydrobiopterin (BH 4 )/ nitric oxide synthase (NOS) expression in primary human aortic endothelial cells (pHAECs). Methods: pHAECs were infected for 48 hours with Pg in vitro using the Human oxygen-Bacteria anaerobic coculture technique. Cell viability was determined, and target gene expression changes were evaluated by quantitative real-time polymerase chain reaction at the end of each incubation period.Results: Pg impaired pHAEC viability 24 hours post-infection. Pg infection reduced mRNA expression levels of endothelial NOS (eNOS), Nrf2, and Phase II enzymes (heme oxygenase-1, catalase, superoxide dismutase-1) in a timedependent manner. Significant (P <0.05) increase in the inflammatory markers (interleukin [IL]-1β, IL-6, and tumor necrosis factor-α) were observed in the medium as well as in the infected cells. Interestingly, inducible NOS mRNA levels showed a significant (P <0.05) increase at 12 hours and 24 hours and were reduced at later time points. BH 4 (cofactor of eNOS) biosynthesis enzyme dihydrofolate reductase (DHFR, salvage pathway) mRNA levels showed a significant (P <0.05) decrease, while mRNA levels of GSK-3β were elevated. Conclusions: These results suggest that periodontal bacterial infection may cause significant changes in the endothelial GSK-3β/BH 4 /eNOS/Nrf2 pathways, which may lead to impaired vascular relaxation. Greater understanding of the factors that adversely affect endothelial cell function could contribute to the development of new therapeutic compounds to treat PD-induced vascular diseases.

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“…Of note, a variety of species, mostly with periodontal origin were cultivated from atheromatous tissue of endarterectomy patients [74][75][76] . These are promising targets for intervention, specifically P. gingivalis, a gram-negative anaerobe capable of invading a variety of non-phagocytic eukaryotic cells [166][167][168][169][170] . P. gingivalis is a key periodontal pathogen [171] causing inflammation and host tissue destruction.…”

Section: Exploring Cutting-edge Technologies To Discover New Therapeumentioning

confidence: 99%

Atherosclerosis microbiome: upcoming target for vaccine and drug development

Kozarov¹,

Progulske‐Fox²

2020

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Cardiovascular disease (CVD) is the leading cause of morbidity and mortality in adults and is one critical area of the medical sciences. Atherosclerosis is the main underlying pathology and is characterized by chronic inflammation of the arterial walls. The current treatment modalities for CVD target hypertension, hyperlipidemia and hemostasis, and suppress inflammation without directly addressing the origin of inflammation. Thus, many individuals with multiple classic risk factors for CVD do not experience acute ischemic events. Moreover, myocardial infarction and stroke continue to occur in up to two-thirds of all patients. Because many cardiovascular events have not been explained by genetics or other risk factors, and multiple epidemiologic studies have consistently suggested an infectious component, the introduction of entirely novel approaches for diagnostics and treatment that target infections are acutely needed. These complementary novel approaches addressing additional manageable risk factors such as infections will be based on the concept of personalized medicine to control CVD and achieve longevity, while also increasing the quality of life. There are a variety of avenues that could enable such novel approaches. These focus on the discovery and characterization of the infective component of atherosclerosis, the atherosclerosis microbiome. Specifically, we provide an update of the latest developments in the oral microbiome and its relation to CVD.

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<em>Porphyromonas gingivalis</em> as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells

Cited by 9 publications

References 50 publications

Bitter Taste Receptor T2R14 Modulates Gram-Positive Bacterial Internalization and Survival in Gingival Epithelial Cells

Bitter Taste Receptor T2R14 Modulates Gram-Positive Bacterial Internalization and Survival in Gingival Epithelial Cells

Porphyromonas gingivalis infection alters Nrf2‐phase II enzymes and nitric oxide in primary human aortic endothelial cells

Atherosclerosis microbiome: upcoming target for vaccine and drug development

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