Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 646
Filtrar
1.
Methods Mol Biol ; 2210: 143-155, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32815135

RESUMO

OmpA-like proteins located in the outer bacterial membrane are potential virulence factors from the major periodontal pathogens Porphyromonas gingivalis and Tannerella forsythia. Our previous studies have shown that OmpA-like proteins are glycosylated by O-linked N-acetylglucosamine (O-GlcNAc) and are strongly reactive to wheat germ agglutinin (WGA) lectin, which shows sugar specificity to GlcNAc. Utilizing this property, we have developed a separation method for OmpA-like proteins by affinity chromatography using WGA lectin-agarose. The purity of enriched native OmpA-like proteins were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining. More importantly, the purified OmpA-like proteins formed a unique trimeric structure keeping their bioactivity intact. In this chapter, we describe a detailed procedure to separate OmpA-like proteins, which may be used to further progress the biological studies of OmpA-like proteins.


Assuntos
Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Proteínas de Bactérias/isolamento & purificação , Cromatografia de Afinidade/métodos , Porphyromonas gingivalis/química , Tannerella forsythia/química , Proteínas da Membrana Bacteriana Externa/análise , Proteínas de Bactérias/análise , Infecções por Bacteroidaceae/microbiologia , Eletroforese em Gel de Poliacrilamida/métodos , Glicosilação , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Multimerização Proteica , Aglutininas do Germe de Trigo/química
2.
Methods Mol Biol ; 2210: 167-172, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32815137

RESUMO

Butyrate is one of the most harmful metabolic end products found in the oral cavity. Thus, it would be important to characterize the enzymes responsible for production of this metabolite to elucidate the pathogenicity of periodontogenic bacteria. Here, a spectrophotometric assay for butyryl-CoA:acetate CoA transferase activity and gas chromatography-mass spectrometry measurement of butyrate and other short chain fatty acids such as acetate, propionate, isobutyrate, and isovalerate are described.


Assuntos
Acil Coenzima A/metabolismo , Proteínas de Bactérias/metabolismo , Butiratos/metabolismo , Coenzima A-Transferases/metabolismo , Porphyromonas gingivalis/metabolismo , Infecções por Bacteroidaceae/microbiologia , Colorimetria/métodos , Ácidos Graxos Voláteis/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Espectrofotometria/métodos
3.
Methods Mol Biol ; 2210: 215-224, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32815142

RESUMO

Porphyromonas gingivalis is a major pathogen responsible for severe and chronic manifestations of periodontal disease, which is one of the most common infectious disorders of humans. Although human gingival epithelium prevents intrusions by periodontal bacteria, P. gingivalis is able to invade gingival epithelial cells. To study the dynamics and the fate of intracellular P. gingivalis, confocal laser scanning microscopy (CLSM) is a method of choice. Information gained with CLSM contains not only the number of P. gingivalis associated with gingival epithelial cells but also the bacterial localization on/inside the host cells, morphological change of host cells, and physical interaction between the bacteria and host organelle. In this chapter, we describe the protocols for microscopy techniques to morphologically study gingival epithelial cells infected by P. gingivalis.


Assuntos
Infecções por Bacteroidaceae/patologia , Células Epiteliais/patologia , Gengiva/patologia , Doenças Periodontais/patologia , Porphyromonas gingivalis/fisiologia , Infecções por Bacteroidaceae/microbiologia , Técnicas de Cultura de Células/métodos , Linhagem Celular , Células Epiteliais/microbiologia , Gengiva/citologia , Gengiva/microbiologia , Humanos , Microscopia Confocal/métodos , Doenças Periodontais/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Coloração e Rotulagem/métodos
4.
Methods Mol Biol ; 2210: 225-233, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32815143

RESUMO

Chronic periodontitis is the most common periodontitis observed in adults. Recently, its association with systemic diseases such as ischemic heart-brain disease and diabetes has been pointed out. Porphyromonas gingivalis, a major causative bacterium of chronic periodontitis, has properties of adhering to blood vessels and inducing inflammation, and those properties are involved in the induction of vascular inflammation and promotion of atherosclerosis. Therefore, analysis of the interaction of P. gingivalis with vascular endothelial cells will contribute to an understanding of the link between periodontitis and vascular lesions.


Assuntos
Infecções por Bacteroidaceae/patologia , Células Endoteliais/patologia , Periodontite/patologia , Porphyromonas gingivalis/fisiologia , Infecções por Bacteroidaceae/microbiologia , Técnicas de Cultura de Células/métodos , Linhagem Celular , Células Endoteliais/microbiologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Óxido Nítrico/metabolismo , Periodontite/microbiologia , Fator de von Willebrand/metabolismo
5.
Biochim Biophys Acta Mol Basis Dis ; 1867(1): 165991, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33080346

RESUMO

Our previous study demonstrated that IL-10 secreting B (B10) cells alleviate inflammation and bone loss in experimental periodontitis. The purpose of this study is to determine whether antigen-specificity is required for the local infiltration of B10 cells. Experimental periodontitis was induced in the recipient mice by placement of silk ligature with or without the presence of live Porphyromonas gingivalis (P. gingivalis). Donor mice were pre-immunized by intraperitoneal (IP) injection of formalin-fixed P. gingivalis, or PBS as non-immunized control. Spleen B cells were purified and treated with LPS and CpG for 48 h to expand the B10 population in vitro. Fluorescence-labelled B10 cells were transferred into the recipient mice by tail vein injection and were tracked on day 0, 3, 5 and 10 using IVIS Spectrum in vivo imaging system. The number of B10 cells and P. gingivalis-binding B cells were significantly increased after in vitro treatment of LPS and CpG. On day 5, the fluorescence intensity in gingival tissues was the highest in mice transferred with B10 cells from pre-immunized donor mice. Gingival expression of IL-6, TNF-α, RANKL/OPG ratio and periodontal bone loss in recipient mice were significantly reduced, and the expression of IL-10 and the number of CD19+ B cells were significantly increased after pre-immunized B10 cell transfer in the presence of antigen, compared to those with non-immunized B10 cell transfer or no antigen presence. This study suggests that antigen specificity dictate the local infiltration of B10 cells into periodontal tissue and these antigen-specific B10 cells promote anti-inflammatory responses.


Assuntos
Antígenos de Bactérias/imunologia , Linfócitos B Reguladores/imunologia , Infecções por Bacteroidaceae , Gengiva , Periodontite , Porphyromonas gingivalis/imunologia , Animais , Infecções por Bacteroidaceae/diagnóstico por imagem , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Citocinas/imunologia , Diagnóstico por Imagem , Gengiva/diagnóstico por imagem , Gengiva/imunologia , Gengiva/microbiologia , Camundongos , Periodontite/diagnóstico por imagem , Periodontite/imunologia , Periodontite/microbiologia
6.
PLoS One ; 15(9): e0238425, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32960889

RESUMO

OBJECTIVE: To evaluate the effects of Bifidobacterium animalis subsp. lactis HN019 (HN019) on clinical periodontal parameters (plaque accumulation and gingival bleeding), on immunocompetence of gingival tissues [expression of beta-defensin (BD)-3, toll-like receptor 4 (TLR4), cluster of differentiation(CD)-57 and CD-4], and on immunological properties of saliva (IgA levels) in non-surgical periodontal therapy in generalized chronic periodontitis (GCP) patients. Adhesion to buccal epithelial cells (BEC) and the antimicrobial properties of HN019 were also investigated. MATERIALS AND METHODS: Thirty patients were recruited and monitored clinically at baseline (before scaling and root planing-SRP) and after 30 and 90 days. Patients were randomly assigned to Test (SRP+Probiotic, n = 15) or Control (SRP+Placebo, n = 15) group. Probiotic lozenges were used for 30 days. Gingival tissues and saliva were immunologically analyzed. The adhesion of HN019 with or without Porphyromonas gingivalis in BEC and its antimicrobial properties were investigated in in vitro assays. Data were statistically analyzed (p<0.05). RESULTS: Test group presented lower plaque index (30 days) and lower marginal gingival bleeding (90 days) when compared with Control group. Higher BD-3, TLR4 and CD-4 expressions were observed in gingival tissues in Test group than in Control group. HN019 reduced the adhesion of P. gingivalis to BEC and showed antimicrobial potential against periodontopathogens. CONCLUSION: Immunological and antimicrobial properties of B. lactis HN019 make it a potential probiotic to be used in non-surgical periodontal therapy of patients with GCP. CLINICAL RELEVANCE: B. lactis HN019 may be a potential probiotic to improve the effects of non-surgical periodontal therapy. Name of the registry and registration number (ClinicalTrials.gov): "Effects of probiotic therapy in the treatment of periodontitis"-NCT03408548.


Assuntos
Bifidobacterium animalis/imunologia , Periodontite Crônica/terapia , Probióticos/uso terapêutico , Adulto , Aderência Bacteriana/imunologia , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Infecções por Bacteroidaceae/terapia , Periodontite Crônica/imunologia , Periodontite Crônica/microbiologia , Método Duplo-Cego , Feminino , Interações entre Hospedeiro e Microrganismos/imunologia , Humanos , Imunoglobulina A Secretora/metabolismo , Técnicas In Vitro , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/imunologia , Mucosa Bucal/microbiologia , Porphyromonas gingivalis/patogenicidade , Saliva/imunologia
7.
Mikrobiyol Bul ; 54(2): 246-256, 2020 Apr.
Artigo em Turco | MEDLINE | ID: mdl-32723280

RESUMO

Prevotella species, being members of the human microbiota, are obligate anaerobic gram-negative bacteria. These organisms may cause opportunistic infections, including specific oral infections, local or systemic infections. A significant increase of resistance to some antimicrobials has been detected among Prevotella species. The frequency of resistance vary among isolates from different infection sources and between geographic locations. The knowledge about the antimicrobial susceptibility patterns of different Prevotella species is limited in Turkey. Providing the antimicrobial susceptibility data of these bacteria is very important for effective empirical treatment. In this study, we aimed to determine susceptibility data for 12 antimicrobial agents against Prevotella strains originating from human infections, collected in two centers in Turkey. A total of 118 Prevotella strains, isolated from different clinical samples in Marmara University Faculty of Medicine Medical Microbiology and Istanbul University Faculty of Dentistry Oral Microbiology Laboratories between January 2014-December 2017, were tested. Organisms were identified by using MALDI-TOF MS and by 16S rRNA gene sequencing. Minimal inhibitor concentrations of ampicillin, ampicillin-sulbactam, piperacillin-tazobactam, cefoxitin, meropenem, imipenem, clindamycin, tetracycline, tigecycline, moxifloxacin and metronidazole were determined using gradiyent test methodology (E-test; bioMerieux, France) and the European Committee on Antimicrobial Susceptibility Testing, Clinical and Laboratory Standards Institute and Food and Drug Administration guidelines were used for interpretation. Thirteen different Prevotella species were identified, Prevotella bivia and Prevotella nigrescens were the most prevalent species (n= 21) followed by Prevotella buccae (n= 19). All Prevotella strains were susceptible to piperacillin-tazobactam, cefoxitin, meropenem, imipenem and tigecycline. A total of 2 (1.7%) isolates were resistant to metronidazole and 1 (0.8%) isolate was intermediately resistant to ampicillin/sulbactam. The frequency of resistant isolates against ampicillin, clindamycin, tetracycline and moxifloxacin were 57.6%, 36.4%, 18% and 16.3%, respectively. In conclusion, piperacillin/tazobactam, cefoxitin, and tigecycline displayed high in vitro activity against Prevotella spp. and they all remained good candidates for empiric therapy. Imipenem and meropenem were also found to be very active, but the usage of carbapenems should be reserved for serious mixed infections, potentially accompanied by other resistant organisms. Intermediate resistance to ampicillinsulbactam and the resistance against metronidazole emphasized the need of periodic monitoring of their susceptibility patterns. The high rates of non-susceptibility to ampicillin, clindamycin, tetracycline and moxifloxacin indicated that these antimicrobials should not be used for treatment of infections without prior antimicrobial susceptibility testing.


Assuntos
Bactérias Anaeróbias , Prevotella , Antibacterianos/farmacologia , Infecções por Bacteroidaceae/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Prevotella/efeitos dos fármacos , Prevotella/genética , RNA Ribossômico 16S/genética , Turquia
8.
Ann Rheum Dis ; 79(9): 1194-1202, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32532752

RESUMO

OBJECTIVES: Porphyromonas gingivalis (P.g.) is discussed to be involved in triggering self-reactive immune responses. The aim of this study was to investigate the autocitrullinated prokaryotic peptidylarginine deiminase (PPAD) from P.g. CH2007 (RACH2007-PPAD) from a rheumatoid arthritis (RA) patient and a synthetic citrullinated PPAD peptide (CPP) containing the main autocitrullination site as potential targets for antibody reactivity in RA and to analyse the possibility of citrullinating native human proteins by PPAD in the context of RA. METHODS: Recombinant RACH2007-PPAD was cloned and expressed in Escherichia coli. Purified RACH2007-PPAD and its enzymatic activity was analysed using two-dimensional electrophoresis, mass spectrometry, immunoblot and ELISA. Autoantibody response to different modified proteins and peptides was recorded and bioinformatically evaluated. RESULTS: RACH2007-PPAD was capable to citrullinate major RA autoantigens, such as fibrinogen, vimentin, hnRNP-A2/B1, histone H1 and multiple peptides, which identify a common RG/RGG consensus motif. 33% of RA patients (n=30) revealed increased reactivity for α-cit-RACH2007-PPAD before RA onset. 77% of RA patients (n=99) presented α-cit-specific signals to CPP amino acids 57-71 which were positively correlated to α-CCP2 antibody levels. Interestingly, 48% of the α-CPP-positives were rheumatoidfactor IgM/anti-citrullinated peptide/protein antibodies (ACPA)-negative. Anti-CPP and α-RACH2007-PPAD antibody levels increase with age. Protein macroarrays that were citrullinated by RACH2007-PPAD and screened with RA patient sera (n=6) and controls (n=4) uncovered 16 RACH2007-PPAD citrullinated RA autoantigens and 9 autoantigens associated with lung diseases. We showed that the α-CPP response could be an important determinant in parenchymal changes in the lung at the time of RA diagnosis (n=106; p=0.018). CONCLUSIONS: RACH2007-PPAD induced internal citrullination of major RA autoantigens. Anti-RACH2007-PPAD correlates with ACPA levels and interstitial lung disease autoantigen reactivity, supporting an infection-based concept for induction of ACPAs via enzymatic mimicry.


Assuntos
Anticorpos Anti-Proteína Citrulinada/imunologia , Artrite Reumatoide/imunologia , Autoanticorpos/imunologia , Infecções por Bacteroidaceae/imunologia , Epitopos/imunologia , Porphyromonas gingivalis/imunologia , Artrite Reumatoide/microbiologia , Infecções por Bacteroidaceae/microbiologia , Citrulinação/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Immunoblotting , Peptídeos/imunologia , Desiminases de Arginina em Proteínas/imunologia
9.
PLoS Pathog ; 16(6): e1008559, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32497109

RESUMO

Antibiotics continue to be the standard-of-care for bacterial vaginosis (BV), although recurrence rates are high. Vaginal probiotics may improve durability of BV treatment, although few probiotics for vaginal health contain Lactobacillus spp. that commonly colonize the lower female genital tract. Characteristics of vaginal Lactobacillus strains from South African women were evaluated for their probiotic potential in vitro compared to strains from commercial vaginal products, including growth at varying pHs, ability to lower pH, produce D-/L-lactate and H2O2, influence growth of BV-associated Gardnerella vaginalis and Prevotella bivia, adherence to cervical cells and susceptibility to antibiotics. Fifty-seven Lactobacillus strains were purified from cervico-vaginal fluid, including L. crispatus, L. jensenii, L. gasseri, L. mucosae, and L. vaginalis. L crispatus strains grew better at pHs below 4.5 and lowered pH more effectively than other strains. Production of D-/L-lactate and H2O2 varied between Lactobacillus species and strains. Lactobacillus strains generally inhibited P. bivia more uniformly than G. vaginalis isolates. All vaginal Lactobacillus isolates were resistant to metronidazole while susceptibility to clindamycin varied. Furthermore, vaginal Lactobacillus strains tended to be broadly susceptible to penicillin, amoxicillin, rifampicin and rifabutin. Whole-genome-sequencing of five of the best-performing vaginal Lactobacillus strains confirmed their likely safety, due to antimicrobial resistance elements being largely absent, while putative intact prophages were present in the genomes of two of the five strains. Overall, vaginal Lactobacillus strains largely performed better in these in vitro assays than probiotic strains currently used in probiotics for vaginal health. Including the best-performing vaginal Lactobacillus isolates in a region-specific probiotic for vaginal health may result in improved BV treatment options.


Assuntos
Infecções por Bacteroidaceae/microbiologia , Gardnerella vaginalis , Infecções por Bactérias Gram-Positivas/microbiologia , Lactobacillus , Prevotella , Vaginose Bacteriana/microbiologia , Adolescente , Adulto , Infecções por Bacteroidaceae/tratamento farmacológico , Infecções por Bacteroidaceae/genética , Infecções por Bacteroidaceae/metabolismo , Clindamicina/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Farmacorresistência Bacteriana/genética , Feminino , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/genética , Infecções por Bactérias Gram-Positivas/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Ácido Láctico/metabolismo , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Metronidazol/farmacologia , África do Sul , Especificidade da Espécie , Vaginose Bacteriana/tratamento farmacológico , Vaginose Bacteriana/genética
10.
Sci Rep ; 10(1): 7468, 2020 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-32366945

RESUMO

Recent epidemiological  studies link Periodontal disease(PD) to age-related macular degeneration (AMD). We documented earlier that Porphyromonas gingivalis(Pg), keystone oral-pathobiont, causative of PD, efficiently invades human gingival epithelial and blood-dendritic cells. Here, we investigated the ability of dysbiotic Pg-strains to invade human-retinal pigment epithelial cells(ARPE-19), their survival, intracellular localization, and the pathological effects, as dysfunction of RPEs leads to AMD. We show that live, but not heat-killed Pg-strains adhere to and invade ARPEs. This involves early adhesion to ARPE cell membrane, internalization and localization of Pg within single-membrane vacuoles or cytosol, with some nuclear localization apparent. No degradation of Pg or localization inside double-membrane autophagosomes was evident, with dividing Pg suggesting a metabolically active state during invasion. We found significant downregulation of autophagy-related genes particularly, autophagosome complex. Antibiotic protection-based recovery assay further confirmed distinct processes of adhesion, invasion and amplification of Pg within ARPE cells. This is the first study to demonstrate invasion of human-RPEs, begin to characterize intracellular localization and survival of Pg within these cells. Collectively, invasion of RPE by Pg and its prolonged survival by autophagy evasion within these cells suggest a strong rationale for studying the link between oral infection and AMD pathogenesis in individuals with periodontitis.


Assuntos
Autofagossomos , Autofagia , Infecções por Bacteroidaceae , Citosol , Porphyromonas gingivalis , Epitélio Pigmentado da Retina , Vacúolos , Autofagossomos/metabolismo , Autofagossomos/microbiologia , Autofagossomos/ultraestrutura , Infecções por Bacteroidaceae/metabolismo , Infecções por Bacteroidaceae/microbiologia , Infecções por Bacteroidaceae/patologia , Linhagem Celular , Citosol/metabolismo , Citosol/microbiologia , Citosol/ultraestrutura , Humanos , Porphyromonas gingivalis/metabolismo , Porphyromonas gingivalis/ultraestrutura , Epitélio Pigmentado da Retina/metabolismo , Epitélio Pigmentado da Retina/microbiologia , Epitélio Pigmentado da Retina/ultraestrutura , Vacúolos/microbiologia , Vacúolos/patologia , Vacúolos/ultraestrutura
11.
Infect Immun ; 88(5)2020 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-32041789

RESUMO

Programmed death-ligand 1 (PD-L1/B7-H1) serves as a cosignaling molecule in cell-mediated immune responses and contributes to chronicity of inflammation and the escape of tumor cells from immunosurveillance. Here, we investigated the molecular mechanisms leading to PD-L1 upregulation in human oral carcinoma cells and in primary human gingival keratinocytes in response to infection with Porphyromonas gingivalis (P. gingivalis), a keystone pathogen for the development of periodontitis. The bacterial cell wall component peptidoglycan uses bacterial outer membrane vesicles to be taken up by cells. Internalized peptidoglycan triggers cytosolic receptors to induce PD-L1 expression in a myeloid differentiation primary response 88 (Myd88)-independent and receptor-interacting serine/threonine-protein kinase 2 (RIP2)-dependent fashion. Interference with the kinase activity of RIP2 or mitogen-activated protein (MAP) kinases interferes with inducible PD-L1 expression.


Assuntos
Antígeno B7-H1/metabolismo , Infecções por Bacteroidaceae/metabolismo , Carcinoma/metabolismo , Parede Celular/metabolismo , Neoplasias Bucais/metabolismo , Porphyromonas gingivalis/metabolismo , Proteína Serina-Treonina Quinase 2 de Interação com Receptor/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Infecções por Bacteroidaceae/microbiologia , Carcinoma/microbiologia , Linhagem Celular Tumoral , Gengiva/metabolismo , Gengiva/microbiologia , Humanos , Queratinócitos/metabolismo , Queratinócitos/microbiologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Neoplasias Bucais/microbiologia , Periodontite/metabolismo , Periodontite/microbiologia , Regulação para Cima/fisiologia
12.
Support Care Cancer ; 28(10): 4729-4735, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31965308

RESUMO

PURPOSE: Clinical and in vitro studies showed selected oral microorganisms to be related to delayed wound healing and ulcerative oral mucositis. However, it is not known whether this effect is due to reduced metabolism and/or the reduced reproductive capacity of epithelial cells. Therefore, we studied the influence of the oral microorganisms Porphyromonas gingivalis, Candida glabrata, and Candida kefyr on cell metabolism and reproductive capacity of oral epithelial cells, aimed to further unravel the pathogenesis of oral mucositis. METHODS: Oral epithelial cells were exposed to different concentrations of P. gingivalis, C. glabrata, and C. kefyr as mono-infections or mixed together. An MTT assay was performed to determine the effect on cell metabolism. A clonogenic assay was used to study the effect on the reproductive capacity of oral epithelial cells. RESULTS: The metabolism of oral epithelial cells was reduced when the microorganisms were present in high concentrations: P. gingivalis at a multiplicity of infection (MOI) of 1000 and the Candida spp. at MOI 100. No statistical difference was observed in the ability of a single epithelial cell to grow into a colony of cells between control and P. gingivalis, C. glabrata, and C. kefyr, independent of the concentrations and combinations used. CONCLUSION: P. gingivalis, C. glabrata, and C. kefyr lowered the metabolic activity of oral epithelial cells in high concentrations, yet they did not influence the reproductive capacity of epithelial cells. Their impact on ulcerative oral mucositis is likely due to an effect on the migration, proliferation, and metabolism of epithelial cells.


Assuntos
Candida/fisiologia , Porphyromonas gingivalis/fisiologia , Estomatite/microbiologia , Infecções por Bacteroidaceae/metabolismo , Infecções por Bacteroidaceae/microbiologia , Infecções por Bacteroidaceae/patologia , Candida glabrata/fisiologia , Candidíase/metabolismo , Candidíase/microbiologia , Candidíase/patologia , Linhagem Celular , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Humanos , Técnicas In Vitro , Estomatite/metabolismo , Estomatite/patologia
13.
Nutrients ; 12(1)2020 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-31968635

RESUMO

Periodontitis is a polymicrobial infectious disease that leads to inflammation of the gingiva, resulting in teeth loss by various causes such as inflammation-mediated bone resorption. Recently, many investigators have reported that the periodontitis resulting from persistent low-grade infection of Gram-negative bacteria such as Porphyromonas gingivalis (Pg) is associated with increased atherosclerosis, diabetes mellitus, and other systemic diseases through blood stream. On the other hand, carotenoids belong among phytochemicals that are responsible for different colors of the foods. It is important to examine whether carotenoids are effective to the inhibition of periodontal infection/inflammation cascades. This review summarizes the advanced state of knowledge about suppression of periodontal infection by several carotenoids. A series of findings suggest that carotenoids intake may provide novel strategy for periodontitis treatment, although further study will be needed.


Assuntos
Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Infecções por Bacteroidaceae/tratamento farmacológico , Carotenoides/uso terapêutico , Periodontite/tratamento farmacológico , Porphyromonas gingivalis/patogenicidade , Animais , Anti-Inflamatórios/efeitos adversos , Antioxidantes/efeitos adversos , Infecções por Bacteroidaceae/microbiologia , Carotenoides/efeitos adversos , Humanos , Mediadores da Inflamação/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Periodontite/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Resultado do Tratamento
14.
Anaerobe ; 61: 102140, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31838319

RESUMO

Porphyromonas gingivalis is a keystone pathogen in periodontitis. Analysis of the immunogenicity of its virulence factors may provide insight into the host response to this infection. The Kgp12 (IEDB Epitope ID 763561), an epitope of Lys-gingipain (Kgp) virulence factor from P. gingivalis ATCC 33277, elicits an immunoglobulin G (IgG) immunoreactivity with low cross-reactivity and, therefore, more specificity. The aim of the present study was to determine in silico the localization of Kgp12 within the protein and to evaluate the IgG host response to this novel Kgp peptide through its capacity to differentiate individuals with different periodontal status. Sera of 71 volunteers were tested by indirect ELISA to detect the IgG immunoreactivity specific to Kgp12, as well as to the protein HmuY and to the sonicated total extract of P. gingivalis ATCC33277, both used as gold standard. The participants had no systemic disease and were classified according to periodontal clinical parameters to comparison, firstly, into periodontitis (P) and without periodontitis (WP) groups and, secondly, into periodontitis (P), gingivitis (G) and clinically health (CH) ones. All the antigens tested, Kgp12 (p = 0.02), HmuY (p = 0.00) and P. gingivalis extract (p = 0.03), could differentiate P from WP groups considering IgG serum levels. P group also had higher IgG levels specific to Kgp12 (p = 0.03), HmuY (p < 0.01) and P. gingivalis extract (p = 0.01) when compared to G group. We conclude that the Kgp12 synthetic peptide was useful to detect the IgG-mediated host response signaling that it is a promising epitope to analyze the immunogenicity of P. gingivalis.


Assuntos
Infecções por Bacteroidaceae/metabolismo , Infecções por Bacteroidaceae/microbiologia , Cisteína Endopeptidases Gingipaínas/metabolismo , Imunoglobulina G/imunologia , Fragmentos de Peptídeos/metabolismo , Periodontite/etiologia , Porphyromonas gingivalis/enzimologia , Infecções por Bacteroidaceae/imunologia , Bases de Dados de Proteínas , Suscetibilidade a Doenças , Epitopos/imunologia , Feminino , Cisteína Endopeptidases Gingipaínas/química , Cisteína Endopeptidases Gingipaínas/imunologia , Humanos , Imunoglobulina G/sangue , Masculino , Modelos Moleculares , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/imunologia , Porphyromonas gingivalis/imunologia , Transporte Proteico , Relação Estrutura-Atividade
15.
Biochem Biophys Res Commun ; 522(1): 184-190, 2020 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-31757417

RESUMO

Metabolic reprogramming from oxidative phosphorylation to glycolysis have been implicated in the pathogenesis of inflammatory diseases, such as pulmonary hypertension, rheumatoid arthritis and sepsis. Whether metabolic reprogramming participates in the progression of bacteriogenic periodontitis has never been reported. In the present study, we explored metabolic changes in periodontal ligament cells (PDLSCs) in response to Porphyromonas gingivalis. (P. gingivalis)-infected PDLSCs showed distinct metabolomics with metabolic reprogramming from oxidative phosphorylation to glycolysis. In addition, bacteria invasion triggered fundamental changes in glycolysis and tricarboxylate acid (TCA) cycle-related genes, such as the hexokinase (HK), isocitrate dehydrogenase (IDH) and succinate dehydrogenase (SDH). Moreover, P. gingivalis-infected PDLSCs showed accumulation of succinate, elevation in succinate dehydrogenase activity, pileup of reactive oxygen species and activation of hypoxia inducible factor-1α (HIF-1α) pathway. HIF-1α and succinate inhibitors, as well as SDH knockdown alleviated proinflammatory cytokine expression in P. gingivalis-infected PDLSCs. Therefore, targeting metabolic reprogramming by regulating the succinate-SDH-HIF-1α axis may facilitate host modulation therapy of chronic periodontitis.


Assuntos
Infecções por Bacteroidaceae/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Ligamento Periodontal/metabolismo , Periodontite/metabolismo , Porphyromonas gingivalis/fisiologia , Succinato Desidrogenase/metabolismo , Infecções por Bacteroidaceae/microbiologia , Células Cultivadas , Glicólise , Interações Hospedeiro-Patógeno , Humanos , Inflamação/metabolismo , Inflamação/microbiologia , Fosforilação Oxidativa , Ligamento Periodontal/citologia , Ligamento Periodontal/microbiologia , Periodontite/microbiologia , Transdução de Sinais , Ácido Succínico/metabolismo
16.
Immunol Lett ; 218: 11-21, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31863783

RESUMO

Aging humans display an increased prevalence and severity of periodontitis, although the mechanisms underlying these findings remain poorly understood. This report examined antigenic diversity of P. gingivalis related to disease presence and patient demographics. Serum IgG antibody to P. gingivalis strains ATCC33277, FDC381, W50 (ATCC53978), W83, A7A1-28 (ATCC53977) and A7436 was measured in 426 participants [periodontally healthy (n = 61), gingivitis (N = 66) or various levels of periodontitis (N = 299)]. We hypothesized that antigenic diversity in P. gingivalis could contribute to a lack of "immunity" in the chronic infections of periodontal disease. Across the strains, the antibody levels in the oldest age group were lower than in the youngest groups, and severe periodontitis patients did not show higher antibody with aging. While 80 % of the periodontitis patients in any age group showed an elevated response to at least one of the P. gingivalis strains, the patterns of individual responses in the older group were also substantially different than the other age groups. Significantly greater numbers of older patients showed strain-specific antibody profiles to only 1 strain. The findings support that P. gingivalis may demonstrate antigenic diversity/drift within patients and could be one factor to help explain the inefficiency/ineffectiveness of the adaptive immune response in managing the infection.


Assuntos
Anticorpos Antibacterianos/imunologia , Infecções por Bacteroidaceae/diagnóstico , Infecções por Bacteroidaceae/imunologia , Variação Biológica Individual , Periodontite/diagnóstico , Periodontite/etiologia , Porphyromonas gingivalis/imunologia , Adulto , Idoso , Anticorpos Antibacterianos/sangue , Infecções por Bacteroidaceae/microbiologia , Biomarcadores , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Vigilância da População , Adulto Jovem
17.
Acta Microbiol Immunol Hung ; 67(1): 6-13, 2019 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-31813262

RESUMO

In this multicenter study, we aimed to evaluate the performance of MALDI Biotyper and VITEK MS, for identification of Prevotella species. Three hundred and fourteen clinical isolates, collected in eight European countries between January 2014 and April 2016, were identified at the collecting sites by MALDI Biotyper (versions 3.0 and 3.1) and then reidentified by VITEK MS (version 3.0) in the central laboratory. 16S rRNA gene sequencing was used as a standard method. According to sequence analysis, the 314 Prevotella strains belonged to 19 species. MALDI Biotyper correctly identified 281 (89.5%) isolates to the species level and 33 (10.5%) only at the genus level. VITEK MS correctly identified 253 (80.6%) isolates at the species level and 276 (87.9%) isolates at the genus level. Thirty-three isolates belonging to P. bergensis, P. conceptionensis, P. corporis, P. histicola, and P. nanciensis, unavailable in the VITEK MS 3.0 database, were resulted in genus level or no identification. Six Prevotella strains, belonged to P. veroralis, P. timonensis, and P. conceptionensis not represented in the MALDI Biotyper system database, were misidentified at the genus level. In conclusion, both VITEK MS and MALDI Biotyper provided reliable and rapid identification. However, the permanent extension of the databases is needed.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Prevotella/química , Prevotella/classificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Infecções por Bacteroidaceae/microbiologia , Europa (Continente) , Humanos , Análise de Sequência de DNA
18.
Clin Exp Dent Res ; 5(5): 497-504, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31687183

RESUMO

Objectives: Our study investigated the pathological outcome of experimental thrombi that incorporate oral bacteria. Material and methods: A small artery and vein in the rats' groins were injected with a solution containing periodontal bacteria Porphyromonas gingivalis and followed up for 28 days. In all, 18 limbs of nine male rats (500-650 g) were used for the arterial study, and eight limbs of four rats were used for the veins. Two densities of the bacterial solution and two arterial thicknesses sizes were used in the arterial study. Both proximal and distal arteries and veins were ligated loosely using a monofilament nylon suture before bacterial suspensions or control solutions were injected into the ligated vessels. Results: After 7, 14-18, and 28 days, the rats were sacrificed. Pathology and immunohistochemistry were performed. All specimens exhibited thrombus formation and an acute inflammation reaction with granulocytes at 7 days and then settled down to chronic fibrous change with plasma cells or macrophages at 28 days in the arterial thrombus. CD3 (Pan T-cells), CD79a (Pan B cells in the rats), and IgG were observed in the process of the healing of the arterial thrombus. Venous changes showed relatively clear recanalization that appeared at 7 days, which is slightly different from the artery. Granulocytes were present from 7 to 28 days. Conclusions: Periodontal bacteria act as an inflammatory core in the vessels, but not as an infectious agent, in our experiments, because of their low ability to invade tissues.


Assuntos
Artérias/imunologia , Artérias/patologia , Infecções por Bacteroidaceae/complicações , Trombose/imunologia , Trombose/patologia , Veias/imunologia , Veias/patologia , Animais , Artérias/microbiologia , Infecções por Bacteroidaceae/microbiologia , Masculino , Porphyromonas gingivalis/isolamento & purificação , Ratos , Trombose/microbiologia , Veias/microbiologia
19.
Int J Med Sci ; 16(10): 1320-1327, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31692996

RESUMO

Porphyromonas gingivalis is a pivotal periodontal pathogen, and the epithelial cells serve as the first physical barrier to defend the host from bacterial attack. Within this host-bacteria interaction, P. gingivalis can modify the host immune reaction and adjust the gene expression, which is associated with periodontitis pathogenesis and developing strategies. Herein, a meta-analysis was made to get the differential gene expression profiles in epithelial cells with or without P. gingivalis infection. The network-based meta-analysis program for gene expression profiling was used. Both the gene ontology analysis and the pathway enrichment analysis of the differentially expressed genes were conducted. Our results determined that 290 genes were consistently up-regulated in P. gingivalis infected epithelial cells. 229 gene ontology biological process terms of up-regulated genes were discovered, including "negative regulation of apoptotic process" and "positive regulation of cell proliferation/migration/angiogenesis". In addition to the well-known inflammatory signaling pathways, the pathway associated with a transcriptional misregulation in cancer has also been increased. Our findings indicated that P. gingivalis benefited from the survival of epithelial cells, and got its success as a colonizer in oral epithelium. The results also suggested that infection of P. gingivalis might contribute to oral cancer through chronic inflammation. Negative regulation of the apoptotic process and transcriptional misregulation in cancer pathway are important contributors to the cellular physiology changes during infection development, which have particular relevance to the pathogenesis and progressions of periodontitis, even to the occurrence of oral cancer.


Assuntos
Infecções por Bacteroidaceae/imunologia , Interações Hospedeiro-Patógeno/genética , Neoplasias Bucais/patologia , Periodontite/imunologia , Porphyromonas gingivalis/imunologia , Infecções por Bacteroidaceae/genética , Infecções por Bacteroidaceae/microbiologia , Infecções por Bacteroidaceae/patologia , Sobrevivência Celular/genética , Sobrevivência Celular/imunologia , Progressão da Doença , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Perfilação da Expressão Gênica , Ontologia Genética , Gengiva/citologia , Gengiva/imunologia , Gengiva/microbiologia , Interações Hospedeiro-Patógeno/imunologia , Humanos , Mucosa Bucal/citologia , Mucosa Bucal/imunologia , Mucosa Bucal/microbiologia , Neoplasias Bucais/genética , Neoplasias Bucais/imunologia , Neoplasias Bucais/microbiologia , Periodontite/genética , Periodontite/microbiologia , Periodontite/patologia , Porphyromonas gingivalis/isolamento & purificação , Porphyromonas gingivalis/patogenicidade , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Regulação para Cima
20.
PLoS Pathog ; 15(11): e1008124, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31697789

RESUMO

Porphyromonas gingivalis is a major pathogen in severe and chronic manifestations of periodontal disease, which is one of the most common infections of humans. A central feature of P. gingivalis pathogenicity is dysregulation of innate immunity at the gingival epithelial interface; however, the molecular basis underlying P. gingivalis-dependent abrogation of epithelial barrier function remains unknown. Gingival epithelial cells express junctional adhesion molecule (JAM1), a tight junction-associated protein, and JAM1 homodimers regulate epithelial barrier function. Here we show that Arg-specific or Lys-specific cysteine proteases (gingipains) secreted by P. gingivalis can specifically degrade JAM1 at K134 and R234 in gingival epithelial cells, resulting in permeability of the gingival epithelium to 40 kDa dextran, lipopolysaccharide (LPS), and proteoglycan (PGN). A P. gingivalis strain lacking gingipains was impaired in degradation of JAM1. Knockdown of JAM1 in monolayer cells and a three-dimensional multilayered tissue model also increased permeability to LPS, PGN, and gingipains. Inversely, overexpression of JAM1 in epithelial cells prevented penetration by these agents following P. gingivalis infection. Our findings strongly suggest that P. gingivalis gingipains disrupt barrier function of stratified squamous epithelium via degradation of JAM1, allowing bacterial virulence factors to penetrate into subepithelial tissues.


Assuntos
Infecções por Bacteroidaceae/metabolismo , Moléculas de Adesão Celular/metabolismo , Epitélio/metabolismo , Gengiva/metabolismo , Lipopolissacarídeos/metabolismo , Peptidoglicano/metabolismo , Porphyromonas gingivalis/fisiologia , Receptores de Superfície Celular/metabolismo , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Moléculas de Adesão Celular/genética , Células Cultivadas , Humanos , Imunidade Inata , Proteólise , Receptores de Superfície Celular/genética , Junções Íntimas , Fatores de Virulência
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...