Heat killed Lactobacillus acidophilus mediates Fusobacterium nucleatum induced pro-inflammatory responses in epithelial cells

Abstract Probiotics is widespreadly used nowadays. However, the safety issue with the use of live probiotics is still a matter of contention. In recent years, an expanding body of evidence supports the beneficial role of heat killed probiotics in the maintenance of systemic health, whereas the role of these heat killed bacteria on periodontal health remains unclear. This study aimed to evaluate the effects of heat killed probiotics on periodontal pathogen virulence and associated mechanisms. We demonstrated that heat killed Lactobacillus acidophilus was able to coaggregate with Fusobacterium nucleatum, the bridging bacteria of oral biofilm, and inhibit the adhesion and invasion of F. nucleatum, leading to a subsequent elimination of pro-inflammatory cytokine production in oral epithelial cells. This coaggregation further caused a suppression of the virulence gene fap2 expression in F. nucleatum. Therefore, heat killed L. acidophilus might downregulate the pro-inflammatory cytokine expression in epithelial cells via coaggregation with F. nucleatum and suppression of F. nucleatum fap2 expression, which was the first demonstration that heat killed probiotics modulate periodontal disease pathogenesis via coaggregation. Collectively, this finding provides new evidence that heat killed probiotics might exert beneficial effects to periodontal health by coaggregating with periodontal pathogens and modulating their virulence.

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Inducible Expression of Human β-Defensin 2 byFusobacterium nucleatum in Oral Epithelial Cells: Multiple Signaling Pathways and Role of Commensal Bacteria in Innate Immunity and the Epithelial Barrier

Infection and Immunity ◽

10.1128/iai.68.5.2907-2915.2000 ◽

2000 ◽

Vol 68 (5) ◽

pp. 2907-2915 ◽

Cited By ~ 284

Author(s):

Suttichai Krisanaprakornkit ◽

Janet R. Kimball ◽

Aaron Weinberg ◽

Richard P. Darveau ◽

Brian W. Bainbridge ◽

...

Keyword(s):

Innate Immunity ◽

Cell Wall ◽

Epithelial Cells ◽

Signaling Pathways ◽

Inducible Expression ◽

Periodontal Pathogen ◽

Gingival Tissue ◽

Necrosis Factor Alpha ◽

Tnf Α

ABSTRACT Human gingival epithelial cells (HGE) express two antimicrobial peptides of the β-defensin family, human β-defensin 1 (hBD-1) and hBD-2, as well as cytokines and chemokines that contribute to innate immunity. In the present study, the expression and transcriptional regulation of hBD-2 was examined. HBD-2 mRNA was induced by cell wall extract of Fusobacterium nucleatum, an oral commensal microorganism, but not by that of Porphyromonas gingivalis, a periodontal pathogen. HBD-2 mRNA was also induced by the proinflammatory cytokine tumor necrosis factor alpha (TNF-α) and phorbol myristate acetate (PMA), an epithelial cell activator. HBD-2 mRNA was also expressed in 14 of 15 noninflamed gingival tissue samples. HBD-2 peptide was detected by immunofluorescence in HGE stimulated with F. nucleatum cell wall, consistent with induction of the mRNA by this stimulant. Kinetic analysis indicates involvement of multiple distinct signaling pathways in the regulation of hBD-2 mRNA; TNF-α and F. nucleatum cell wall induced hBD-2 mRNA rapidly (2 to 4 h), while PMA stimulation was slower (∼10 h). In contrast, each stimulant induced interleukin 8 (IL-8) within 1 h. The role of TNF-α as an intermediary in F. nucleatum signaling was ruled out by addition of anti-TNF-α that did not inhibit hBD-2 induction. However, inhibitor studies show that F. nucleatum stimulation of hBD-2 mRNA requires both new gene transcription and new protein synthesis. Bacterial lipopolysaccharides isolated from Escherichia coli andF. nucleatum were poor stimulants of hBD-2, although they up-regulated IL-8 mRNA. Collectively, our findings show inducible expression of hBD-2 mRNA via multiple pathways in HGE in a pattern that is distinct from that of IL-8 expression. We suggest that different aspects of innate immune responses are differentially regulated and that commensal organisms have a role in stimulating mucosal epithelial cells in maintaining the barrier that contributes to homeostasis and host defense.

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GPR110 (ADGRF1) mediates anti-inflammatory effects of N-docosahexaenoylethanolamine

Journal of Neuroinflammation ◽

10.1186/s12974-019-1621-2 ◽

2019 ◽

Vol 16 (1) ◽

Cited By ~ 8

Author(s):

Taeyeop Park ◽

Huazhen Chen ◽

Hee-Yong Kim

Keyword(s):

Inflammatory Cytokine ◽

Inflammatory Responses ◽

Cytokine Expression ◽

Regulatory Function ◽

Enzyme Linked Immunosorbent Assay ◽

Inflammatory Condition ◽

Wild Type ◽

Knock Out ◽

The Brain

Abstract Background Neuroinflammation is a widely accepted underlying condition for various pathological processes in the brain. In a recent study, synaptamide, an endogenous metabolite derived from docosahexaenoic acid (DHA, 22:6n-3), was identified as a specific ligand to orphan adhesion G-protein-coupled receptor 110 (GPR110, ADGRF1). Synaptamide has been shown to suppress lipopolysaccharide (LPS)-induced neuroinflammation in mice, but involvement of GPR110 in this process has not been established. In this study, we investigated the possible immune regulatory role of GPR110 in mediating the anti-neuroinflammatory effects of synaptamide under a systemic inflammatory condition. Methods For in vitro studies, we assessed the role of GPR110 in synaptamide effects on LPS-induced inflammatory responses in adult primary mouse microglia, immortalized murine microglial cells (BV2), primary neutrophil, and peritoneal macrophage by using quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA) as well as neutrophil migration and ROS production assays. To evaluate in vivo effects, wild-type (WT) and GPR110 knock-out (KO) mice were injected with LPS intraperitoneally (i.p.) or TNF intravenously (i.v.) followed by synaptamide (i.p.), and expression of proinflammatory mediators was measured by qPCR, ELISA, and western blot analysis. Activated microglia in the brain and NF-kB activation in cells were examined microscopically after immunostaining for Iba-1 and RelA, respectively. Results Intraperitoneal (i.p.) administration of LPS increased TNF and IL-1β in the blood and induced pro-inflammatory cytokine expression in the brain. Subsequent i.p. injection of the GPR110 ligand synaptamide significantly reduced LPS-induced inflammatory responses in wild-type (WT) but not in GPR110 knock-out (KO) mice. In cultured microglia, synaptamide increased cAMP and inhibited LPS-induced proinflammatory cytokine expression by inhibiting the translocation of NF-κB subunit RelA into the nucleus. These effects were abolished by blocking synaptamide binding to GPR110 using an N-terminal targeting antibody. GPR110 expression was found to be high in neutrophils and macrophages where synaptamide also caused a GPR110-dependent increase in cAMP and inhibition of LPS-induced pro-inflammatory mediator expression. Intravenous injection of TNF, a pro-inflammatory cytokine that increases in the circulation after LPS treatment, elicited inflammatory responses in the brain which were dampened by the subsequent injection (i.p.) of synaptamide in a GPR110-dependent manner. Conclusion Our study demonstrates the immune-regulatory function of GPR110 in both brain and periphery, collectively contributing to the anti-neuroinflammatory effects of synaptamide under a systemic inflammatory condition. We suggest GPR110 activation as a novel therapeutic strategy to ameliorate inflammation in the brain as well as periphery.

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Fusobacterium nucleatum and oral cancer: a critical review

BMC Cancer ◽

10.1186/s12885-021-08903-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Emily McIlvanna ◽

Gerard J. Linden ◽

Stephanie G. Craig ◽

Fionnuala T. Lundy ◽

Jacqueline A. James

Keyword(s):

Oral Cancer ◽

Immune Evasion ◽

Anaerobic Bacterium ◽

Potential Role ◽

Fusobacterium Nucleatum ◽

Squamous Cell Carcinomas ◽

Periodontal Pathogen ◽

Periodontal Pathogens ◽

Cellular Invasion ◽

Oral Carcinogenesis

AbstractThere is a growing level of interest in the potential role inflammation has on the initiation and progression of malignancy. Notable examples include Helicobacter pylori-mediated inflammation in gastric cancer and more recently Fusobacterium nucleatum-mediated inflammation in colorectal cancer. Fusobacterium nucleatum is a Gram-negative anaerobic bacterium that was first isolated from the oral cavity and identified as a periodontal pathogen. Biofilms on oral squamous cell carcinomas are enriched with anaerobic periodontal pathogens, including F. nucleatum, which has prompted hypotheses that this bacterium could contribute to oral cancer development. Recent studies have demonstrated that F. nucleatum can promote cancer by several mechanisms; activation of cell proliferation, promotion of cellular invasion, induction of chronic inflammation and immune evasion. This review provides an update on the association between F. nucleatum and oral carcinogenesis, and provides insights into the possible mechanisms underlying it.

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The Role of CTGF in Inflammatory Responses Induced by Silica Particles in Human Bronchial Epithelial Cells

Lung ◽

10.1007/s00408-019-00272-x ◽

2019 ◽

Vol 197 (6) ◽

pp. 783-791 ◽

Cited By ~ 1

Author(s):

Ting Zhou ◽

Qimei Yu ◽

Hui Lin ◽

Zhenyu Wang ◽

Guoqing Fu ◽

...

Keyword(s):

Epithelial Cells ◽

Inflammatory Responses ◽

Silica Particles ◽

Bronchial Epithelial Cells ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial

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Prototypical anti-inflammatory cytokine IL-10 prevents loss of IGF-I-induced myogenin protein expression caused by IL-1β

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00662.2007 ◽

2008 ◽

Vol 294 (4) ◽

pp. E709-E718 ◽

Cited By ~ 16

Author(s):

Klemen Strle ◽

Robert H. McCusker ◽

Rodney W. Johnson ◽

Samantha M. Zunich ◽

Robert Dantzer ◽

...

Keyword(s):

Inflammatory Cytokine ◽

Muscle Wasting ◽

Inflammatory Responses ◽

Mapk Signaling ◽

Protective Role ◽

Igf I ◽

Anti Inflammatory ◽

Kinase Pathway ◽

Anti Inflammatory Cytokine

Prolonged and excessive inflammation is implicated in resistance to the biological actions of IGF-I and contributes to the pathophysiology of neurodegenerative, metabolic, and muscle-wasting disorders. IL-10 is a critical anti-inflammatory cytokine that restrains inflammatory responses in macrophages and T cells by inhibiting cytokine and chemokine synthesis and reducing expression of their receptors. Here we demonstrate that IL-10 plays a protective role in nonhematopoietic cells by suppressing the ability of exogenous IL-1β to inhibit IGF-I-induced myogenin and myosin heavy chain expression in myoblasts. This action of IL-10 is not caused by impairment of IL-1β-induced synthesis of IL-6 or the ability of IL-1β to activate two members of the MAPK family, ERK1/2 and p38. Instead, this newly defined protective role of IL-10 occurs by specific reversal of IL-1β activation of the JNK kinase pathway. IL-10 blocks IL-1β-induced phosphorylation of JNK, but not ERK1/2 or p38, indicating that only the JNK component of the IL-1β-induced MAPK signaling pathway is targeted by IL-10. This conclusion is supported by the finding that a specific JNK inhibitor acts similarly to IL-10 to restore IGF-I-induced myogenin expression, which is suppressed by IL-1β. Collectively, these data demonstrate that IL-10 acts in a novel, nonclassical, protective manner in nonhematopoietic cells to inhibit the IL-1β receptor-induced JNK kinase pathway, resulting in prevention of IGF-I resistance.

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IL-37 (IL-1F7) the Newest Anti-Inflammatory Cytokine Which Suppresses Immune Responses and Inflammation

International Journal of Immunopathology and Pharmacology ◽

10.1177/039463201202500105 ◽

2012 ◽

Vol 25 (1) ◽

pp. 31-38 ◽

Cited By ~ 43

Author(s):

S. Tetè ◽

D. Tripodi ◽

M. Rosati ◽

F. Conti ◽

G. Maccauro ◽

...

Keyword(s):

Epithelial Cells ◽

Immune Responses ◽

Proinflammatory Cytokines ◽

Mechanism Of Action ◽

Inflammatory Cytokine ◽

Inflammatory Responses ◽

Detailed Mechanism ◽

Anti Inflammatory ◽

And Function ◽

Anti Inflammatory Cytokine

Cytokines such as interleukins, chemokines and interferons are immunomodulating and inflammatory agents, characterized by considerable redundancy, in that many cytokines appear to share similar functions. Virtually all nucleated cells, but especially epithelial cells and macrophages, are potent producers of cytokines. The objective of this study is to review the detailed mechanism of action and the biological profiles of IL-37, the newest anti-inflammatory cytokine. This review focuses on IL-37, a key cytokine in regulating inflammatory responses, mainly by inhibiting the expression, production and function of proinflammatory cytokines: IL-1 family pro-inflammatory effects are markedly suppressed by IL-37.

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Evaluation of Enterococcus faecalis, Staphylococcus warneri and Staphylococcus aureus species in adults with generalized chronic periodontitis

RGO - Revista Gaúcha de Odontologia ◽

10.1590/1981-863720170002000043137 ◽

2017 ◽

Vol 65 (2) ◽

pp. 121-127

Author(s):

Aretuza FRITOLI ◽

Eduardo LOBÃO ◽

Geisla Soares ◽

Belén RETAMAL-VALDES ◽

Magda FERES

Keyword(s):

Staphylococcus Aureus ◽

Enterococcus Faecalis ◽

Chronic Periodontitis ◽

Bacterial Species ◽

Hybridization Technique ◽

Periodontal Pathogens ◽

Periodontal Health ◽

Staphylococcus Warneri ◽

Percentage Points

ABSTRACT Objective: To identify and quantify the levels of three bacterial species that have recently been identified as potential “new” periodontal pathogens (Enterococcus faecalis, Staphylococcus aureus and Staphylococcus warneri) in subjects with periodontal health and generalized chronic periodontitis. Methods: Thirty adults with generalized chronic periodontitis and 10 periodontally healthy were included in this study. Nine subgingival biofilm samples were collected per subject and individually analyzed by checkerboard DNA-DNA hybridization technique. Results: The mean levels of E. faecalis and S. warneri were higher in chronic periodontitis than in periodontal health (p<0.05). Furthermore, a higher percentage of subjects with periodontitis were colonized by the three species evaluated in comparison with healthy subjects (p<0.05). This represented a difference of 40 percentage points between the two groups, for E. faecalis (present in 90% of individuals with periodontitis and 50% of the healthy individuals) and S. warneri (100% and 60%, respectively), and 26 percentage points for S. aureus (86% and 60%, respectively). Conclusion: E. faecalis and S. warneri have the potential to be periodontal pathogens. The role of S. aureus was less evident, since this species was more prevalent and at relatively higher levels in health than the other two species. These data might guide future studies on the role of these microorganisms in the etiology of periodontitis and help to establish more effective treatments for these infections.

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Fusobacterium nucleatum metabolically integrates commensals and pathogens in oral biofilms

10.1101/2021.06.23.449328 ◽

2021 ◽

Author(s):

Akito Sakanaka ◽

Masae Kuboniwa ◽

Shuichi Shimma ◽

Samar A. Alghamdi ◽

Shota Mayumi ◽

...

Keyword(s):

Fusobacterium Nucleatum ◽

Periodontal Pathogen ◽

Oral Microbiota ◽

Periodontal Health ◽

Amino Acid Availability ◽

Metabolic Capability ◽

Oral Biofilms ◽

Health And Disease ◽

Biofilm Development ◽

Polymicrobial Communities

Fusobacterium nucleatum is a common constituent of the oral microbiota in both periodontal health and disease. Previously, we discovered ornithine cross-feeding between F. nucleatum and Streptococcus gordonii, where S. gordonii secretes ornithine via an arginine-ornithine antiporter (ArcD), which in turn supports the growth and biofilm development of F. nucleatum; however, broader metabolic aspects of F. nucleatum within polymicrobial communities and their impact on periodontal pathogenesis have not been addressed. Here, we show that when co-cultured with S. gordonii, F. nucleatum increased amino acid availability to enhance the production of butyrate and putrescine, a polyamine produced by ornithine decarboxylation. Co-culture with Veillonella parvula, another common inhabitant of the oral microbiota, also increased lysine availability, promoting cadaverine production by F. nucleatum. We confirmed that ArcD-dependent ornithine excretion by S. gordonii results in synergistic putrescine production, and mass spectrometry imaging revealed this metabolic capability creates a putrescine-rich microenvironment inside F. nucleatum biofilms. We further demonstrated that polyamines caused significant changes in the biofilm phenotype of a periodontal pathogen, Porphyromonas gingivalis, with putrescine being a potent stimulator of biofilm development and dispersal, and confirmed that F. nucleatum-mediated conversion of ornithine to putrescine enhances biofilm formation by P. gingivalis. Lastly, analysis of plaque samples revealed cooccurrence of P. gingivalis with genetic modules for putrescine production by S. gordonii and F. nucleatum. Overall, our results highlight the ability of F. nucleatum to induce synergistic polyamine production within multi-species consortia, and provide insight into how the trophic web in oral biofilm ecosystems can eventually shape disease-associated communities.

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