Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Nature ; 561(7721): 122-126, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30111836

RESUMO

Immune recognition of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors often activates proinflammatory NF-κB signalling1. Recent studies indicate that the bacterial metabolite D-glycero-ß-D-manno-heptose 1,7-bisphosphate (HBP) can activate NF-κB signalling in host cytosol2-4, but it is unclear whether HBP is a genuine PAMP and the cognate pattern recognition receptor has not been identified. Here we combined a transposon screen in Yersinia pseudotuberculosis with biochemical analyses and identified ADP-ß-D-manno-heptose (ADP-Hep), which mediates type III secretion system-dependent NF-κB activation and cytokine expression. ADP-Hep, but not other heptose metabolites, could enter host cytosol to activate NF-κB. A CRISPR-Cas9 screen showed that activation of NF-κB by ADP-Hep involves an ALPK1 (alpha-kinase 1)-TIFA (TRAF-interacting protein with forkhead-associated domain) axis. ADP-Hep directly binds the N-terminal domain of ALPK1, stimulating its kinase domain to phosphorylate and activate TIFA. The crystal structure of the N-terminal domain of ALPK1 and ADP-Hep in complex revealed the atomic mechanism of this ligand-receptor recognition process. HBP was transformed by host adenylyltransferases into ADP-heptose 7-P, which could activate ALPK1 to a lesser extent than ADP-Hep. ADP-Hep (but not HBP) alone or during bacterial infection induced Alpk1-dependent inflammation in mice. Our findings identify ALPK1 and ADP-Hep as a pattern recognition receptor and an effective immunomodulator, respectively.


Assuntos
Açúcares de Adenosina Difosfato/imunologia , Burkholderia cenocepacia , Citosol , Imunidade Inata , Padrões Moleculares Associados a Patógenos/imunologia , Proteínas Quinases/metabolismo , Yersinia pseudotuberculosis , Açúcares de Adenosina Difosfato/metabolismo , Animais , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/imunologia , Infecções por Burkholderia/patologia , Burkholderia cenocepacia/genética , Burkholderia cenocepacia/imunologia , Burkholderia cenocepacia/metabolismo , Sistemas CRISPR-Cas , Cristalografia por Raios X , Citocinas/biossíntese , Citosol/enzimologia , Citosol/imunologia , Dissacarídeos/metabolismo , Ativação Enzimática , Feminino , Edição de Genes , Fatores Imunológicos/imunologia , Fatores Imunológicos/metabolismo , Imunomodulação , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Moleculares , NF-kappa B/metabolismo , Padrões Moleculares Associados a Patógenos/metabolismo , Yersinia pseudotuberculosis/genética , Yersinia pseudotuberculosis/imunologia , Yersinia pseudotuberculosis/metabolismo
2.
J Med Toxicol ; 13(2): 173-179, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28105575

RESUMO

INTRODUCTION: Bongkrekic acid (BA) has a unique mechanism of toxicity among the mitochondrial toxins: it inhibits adenine nucleotide translocase (ANT) rather than the electron transport chain. Bongkrekic acid is produced by the bacterium Burkholderia gladioli pathovar cocovenenans (B. cocovenenans) which has been implicated in outbreaks of food-borne illness involving coconut- and corn-based products in Indonesia and China. Our objective was to summarize what is known about the epidemiology, exposure sources, toxicokinetics, pathophysiology, clinical presentation, and diagnosis and treatment of human BA poisoning. METHODS: We searched MEDLINE (1946 to present), EMBASE (1947 to present), SCOPUS, The Indonesia Publication Index ( http://id.portalgaruda.org/ ), ToxNet, book chapters, Google searches, Pro-MED alerts, and references from previously published journal articles. We identified a total of 109 references which were reviewed. Of those, 29 (26 %) had relevant information and were included. Bongkrekic acid is a heat-stable, highly unsaturated tricarboxylic fatty acid with a molecular weight of 486 kDa. Outbreaks have been reported from Indonesia, China, and more recently in Mozambique. Very little is known about the toxicokinetics of BA. Bongkrekic acid produces its toxic effects by inhibiting mitochondrial (ANT). ANT can also alter cellular apoptosis. Signs and symptoms in humans are similar to the clinical findings from other mitochondrial poisons, but they vary in severity and time course. Management of patients is symptomatic and supportive. CONCLUSIONS: Bongkrekic acid is a mitochondrial ANT toxin and is reported primarily in outbreaks of food-borne poisoning involving coconut and corn. It should be considered in outbreaks of food-borne illness when signs and symptoms manifest involving the liver, brain, and kidneys and when coconut- or corn-based foods are implicated.


Assuntos
Ácido Bongcréquico/envenenamento , Infecções por Burkholderia/microbiologia , Burkholderia gladioli/metabolismo , Cocos/microbiologia , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Mitocôndrias/enzimologia , Translocases Mitocondriais de ADP e ATP/antagonistas & inibidores , Zea mays/microbiologia , Animais , Ácido Bongcréquico/farmacocinética , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/epidemiologia , Infecções por Burkholderia/terapia , Burkholderia gladioli/patogenicidade , Doenças Transmitidas por Alimentos/enzimologia , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/terapia , Mitocôndrias/patologia , Translocases Mitocondriais de ADP e ATP/metabolismo , Resultado do Tratamento
3.
Cell Host Microbe ; 19(5): 664-74, 2016 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-27133449

RESUMO

Burkholderia cenocepacia is an opportunistic pathogen of the cystic fibrosis lung that elicits a strong inflammatory response. B. cenocepacia employs a type VI secretion system (T6SS) to survive in macrophages by disarming Rho-type GTPases, causing actin cytoskeletal defects. Here, we identified TecA, a non-VgrG T6SS effector responsible for actin disruption. TecA and other bacterial homologs bear a cysteine protease-like catalytic triad, which inactivates Rho GTPases by deamidating a conserved asparagine in the GTPase switch-I region. RhoA deamidation induces caspase-1 inflammasome activation, which is mediated by the familial Mediterranean fever disease protein Pyrin. In mouse infection, the deamidase activity of TecA is necessary and sufficient for B. cenocepacia-triggered lung inflammation and also protects mice from lethal B. cenocepacia infection. Therefore, Burkholderia TecA is a T6SS effector that modifies a eukaryotic target through an asparagine deamidase activity, which in turn elicits host cell death and inflammation through activation of the Pyrin inflammasome.


Assuntos
Proteínas de Bactérias/metabolismo , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/imunologia , Burkholderia cenocepacia/imunologia , Inflamassomos/metabolismo , Pirina/imunologia , Proteínas rho de Ligação ao GTP/imunologia , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/metabolismo , Animais , Infecções por Burkholderia/metabolismo , Burkholderia cenocepacia/enzimologia , Burkholderia cenocepacia/genética , Burkholderia cenocepacia/metabolismo , Caspase 1/metabolismo , Linhagem Celular , Células HEK293 , Humanos , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia/enzimologia , Pneumonia/imunologia , Pirina/metabolismo , Proteínas rho de Ligação ao GTP/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo
4.
PLoS Pathog ; 10(8): e1004327, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25166912

RESUMO

Two distinct defense strategies can protect the host from infection: resistance is the ability to destroy the infectious agent, and tolerance is the ability to withstand infection by minimizing the negative impact it has on the host's health without directly affecting pathogen burden. Burkholderia pseudomallei is a Gram-negative bacterium that infects macrophages and causes melioidosis. We have recently shown that inflammasome-triggered pyroptosis and IL-18 are equally important for resistance to B. pseudomallei, whereas IL-1ß is deleterious. Here we show that the detrimental role of IL-1ß during infection with B. pseudomallei (and closely related B. thailandensis) is due to excessive recruitment of neutrophils to the lung and consequent tissue damage. Mice deficient in the potentially damaging enzyme neutrophil elastase were less susceptible than the wild type C57BL/6J mice to infection, although the bacterial burdens in organs and the extent of inflammation were comparable between C57BL/6J and elastase-deficient mice. In contrast, lung tissue damage and vascular leakage were drastically reduced in elastase-deficient mice compared to controls. Bradykinin levels were higher in C57BL/6 than in elastase-deficient mice; administration of a bradykinin antagonist protected mice from infection, suggesting that increased vascular permeability mediated by bradykinin is one of the mechanisms through which elastase decreases host tolerance to melioidosis. Collectively, these results demonstrate that absence of neutrophil elastase increases host tolerance, rather than resistance, to infection by minimizing host tissue damage.


Assuntos
Infecções por Burkholderia/imunologia , Interações Hospedeiro-Parasita/imunologia , Elastase de Leucócito/imunologia , Elastase de Leucócito/metabolismo , Infecções Respiratórias/imunologia , Animais , Western Blotting , Infecções por Burkholderia/enzimologia , Modelos Animais de Doenças , Citometria de Fluxo , Pulmão/microbiologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infecções Respiratórias/enzimologia , Infecções Respiratórias/microbiologia
5.
J Biol Chem ; 288(42): 30473-84, 2013 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-24014026

RESUMO

AtsR is a membrane-bound hybrid sensor kinase of Burkholderia cenocepacia that negatively regulates quorum sensing and virulence factors such as biofilm production, type 6-secretion, and protease secretion. Here we elucidate the mechanism of AtsR phosphorelay by site-directed mutagenesis of predicted histidine and aspartic acid phosphoacceptor residues. We demonstrate by in vitro phosphorylation that histidine 245 and aspartic acid 536 are conserved sites of phosphorylation in AtsR, and we also identify the cytosolic response regulator AtsT (BCAM0381) as a key component of the AtsR phosphorelay pathway. Monitoring the function of AtsR and its derivatives in vivo by measuring extracellular protease activity and swarming motility confirmed the in vitro phosphorylation results. Together we find that the AtsR receiver domain plays a fine-tuning role in determining the levels of phosphotransfer from its sensor kinase domain to the AtsT response regulator.


Assuntos
Proteínas de Bactérias/metabolismo , Burkholderia cenocepacia/enzimologia , Proteínas Quinases/metabolismo , Percepção de Quorum/fisiologia , Transdução de Sinais/fisiologia , Animais , Proteínas de Bactérias/genética , Sistemas de Secreção Bacterianos/fisiologia , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/genética , Burkholderia cenocepacia/genética , Burkholderia cenocepacia/patogenicidade , Linhagem Celular , Camundongos , Fosforilação/fisiologia , Proteínas Quinases/genética
6.
Science ; 339(6122): 975-8, 2013 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-23348507

RESUMO

Caspases are either apoptotic or inflammatory. Among inflammatory caspases, caspase-1 and -11 trigger pyroptosis, a form of programmed cell death. Whereas both can be detrimental in inflammatory disease, only caspase-1 has an established protective role during infection. Here, we report that caspase-11 is required for innate immunity to cytosolic, but not vacuolar, bacteria. Although Salmonella typhimurium and Legionella pneumophila normally reside in the vacuole, specific mutants (sifA and sdhA, respectively) aberrantly enter the cytosol. These mutants triggered caspase-11, which enhanced clearance of S. typhimurium sifA in vivo. This response did not require NLRP3, NLRC4, or ASC inflammasome pathways. Burkholderia species that naturally invade the cytosol also triggered caspase-11, which protected mice from lethal challenge with B. thailandensis and B. pseudomallei. Thus, caspase-11 is critical for surviving exposure to ubiquitous environmental pathogens.


Assuntos
Caspases/metabolismo , Morte Celular , Citosol/microbiologia , Infecções por Bactérias Gram-Negativas/imunologia , Macrófagos/microbiologia , Vacúolos/microbiologia , Animais , Burkholderia/patogenicidade , Burkholderia/fisiologia , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/imunologia , Infecções por Burkholderia/metabolismo , Burkholderia pseudomallei/patogenicidade , Burkholderia pseudomallei/fisiologia , Infecções por Bactérias Gram-Negativas/enzimologia , Infecções por Bactérias Gram-Negativas/microbiologia , Imunidade Inata , Inflamassomos/metabolismo , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Fagossomos/microbiologia , Salmonelose Animal/enzimologia , Salmonelose Animal/imunologia , Salmonelose Animal/microbiologia , Salmonella typhimurium/patogenicidade , Salmonella typhimurium/fisiologia
7.
Cell Adh Migr ; 6(4): 297-301, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22622109

RESUMO

Phagocytosis is an important component of innate immunity that contributes to the eradication of infectious microorganisms; however, successful bacterial pathogens often evade different aspects of host immune responses. A common bacterial evasion strategy entails the production of toxins and/or effectors that disrupt normal host cell processes and because of their importance Rho-family GTPases are often targeted. Burkholderia cenocepacia, an opportunistic pathogen that has a propensity to infect cystic fibrosis patients, is an example of a pathogenic bacterium that has only recently been shown to disrupt Rho GTPase function in professional phagocytes. More specifically, B. cenocepacia disrupts Rac and Cdc42 seemingly through perturbation of guanine nucleotide exchange factor function. Inactivation of Rac, Cdc42 and conceivably other Rho GTPases seriously compromises phagocyte function.


Assuntos
Infecções por Burkholderia/microbiologia , Burkholderia cenocepacia/fisiologia , Macrófagos/enzimologia , Proteína cdc42 de Ligação ao GTP/metabolismo , Proteínas rac de Ligação ao GTP/metabolismo , Animais , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/imunologia , Humanos , Macrófagos/imunologia , Macrófagos/microbiologia , NADPH Oxidases/metabolismo , Fagocitose , Espécies Reativas de Oxigênio/metabolismo
8.
J Immunol ; 188(10): 5003-11, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22491245

RESUMO

Chronic granulomatous disease (CGD) is an inherited disorder characterized by recurrent life-threatening bacterial and fungal infections. CGD results from defective production of reactive oxygen species by phagocytes caused by mutations in genes encoding the NADPH oxidase 2 (NOX2) complex subunits. Mice with a spontaneous mutation in Ncf1, which encodes the NCF1 (p47(phox)) subunit of NOX2, have defective phagocyte NOX2 activity. These mice occasionally develop local spontaneous infections by Staphylococcus xylosus or by the common CGD pathogen Staphylococcus aureus. Ncf1 mutant mice were more susceptible to systemic challenge with these bacteria than were wild-type mice. Transgenic Ncf1 mutant mice harboring the wild-type Ncf1 gene under the human CD68 promoter (MN(+) mice) gained the expression of NCF1 and functional NOX2 activity specifically in monocytes/macrophages, although minimal NOX2 activity was also detected in some CD11b(+)Ly6G(+) cells defined as neutrophils. MN(+) mice did not develop spontaneous infection and were more resistant to administered staphylococcal infections compared with MN(-) mice. Most strikingly, MN(+) mice survived after being administered Burkholderia cepacia, an opportunistic pathogen in CGD patients, whereas MN(-) mice died. Thus, monocyte/macrophage expression of functional NCF1 protected against spontaneous and administered bacterial infections.


Assuntos
Glicoproteínas de Membrana/biossíntese , Monócitos/enzimologia , Monócitos/microbiologia , NADPH Oxidases/biossíntese , Espécies Reativas de Oxigênio/metabolismo , Infecções Estafilocócicas/prevenção & controle , Animais , Antibacterianos/farmacologia , Infecções por Burkholderia/enzimologia , Infecções por Burkholderia/microbiologia , Infecções por Burkholderia/prevenção & controle , Burkholderia cepacia/imunologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Knockout , Camundongos Transgênicos , Monócitos/imunologia , NADPH Oxidase 2 , NADPH Oxidases/fisiologia , Infecções Estafilocócicas/enzimologia , Infecções Estafilocócicas/microbiologia
9.
Am J Physiol Lung Cell Mol Physiol ; 301(4): L575-86, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21743026

RESUMO

Burkholderia cepacia complex is a group of bacterial pathogens that cause opportunistic infections in cystic fibrosis (CF). The most virulent of these is Burkholderia cenocepacia. Matrix metalloproteinases (MMPs) are upregulated in CF patients. The aim of this work was to examine the role of MMPs in the pathogenesis of B. cepacia complex, which has not been explored to date. Real-time PCR analysis showed that B. cenocepacia infection upregulated MMP-2 and MMP-9 genes in the CF lung cell line CFBE41o- within 1 h, whereas MMP-2, -7, and -9 genes were upregulated in the non-CF lung cell line 16HBE14o-. Conditioned media from both cell lines showed increased MMP-9 activation following B. cenocepacia infection. Conditioned media from B. cenocepacia-infected cells significantly reduced the rate of wound healing in confluent lung epithelia (P < 0.05), in contrast to conditioned media from Pseudomonas aeruginosa-infected cells, which showed predominant MMP-2 activation. Treatment of control conditioned media from both cell lines with the MMP activator 4-aminophenylmercuric acetate (APMA) also resulted in clear activation of MMP-9 and to a much lesser extent MMP-2. APMA treatment of control media also delayed the repair of wound healing in confluent epithelial cells. Furthermore, specific inhibition of MMP-9 in medium from cells exposed to B. cenocepacia completely reversed the delay in wound repair. These data suggest that MMP-9 plays a role in the reduced epithelial repair observed in response to B. cenocepacia infection and that its activation following B. cenocepacia infection contributes to the pathogenesis of this virulent pathogen.


Assuntos
Infecções por Burkholderia/enzimologia , Meios de Cultivo Condicionados/farmacologia , Fibrose Cística/enzimologia , Células Epiteliais/efeitos dos fármacos , Pulmão/enzimologia , Metaloproteinase 9 da Matriz/metabolismo , Cicatrização/efeitos dos fármacos , Infecções por Burkholderia/complicações , Infecções por Burkholderia/microbiologia , Infecções por Burkholderia/patologia , Burkholderia cenocepacia/crescimento & desenvolvimento , Linhagem Celular , Meios de Cultivo Condicionados/química , Fibrose Cística/complicações , Fibrose Cística/microbiologia , Fibrose Cística/patologia , DNA Complementar , Células Epiteliais/citologia , Expressão Gênica , Humanos , Pulmão/microbiologia , Pulmão/patologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 7 da Matriz/genética , Metaloproteinase 7 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Inibidores de Metaloproteinases de Matriz , Acetato de Fenilmercúrio/análogos & derivados , Acetato de Fenilmercúrio/farmacologia , Reação em Cadeia da Polimerase , Inibidores de Proteases/farmacologia , Infecções por Pseudomonas/enzimologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/patologia , Pseudomonas aeruginosa/crescimento & desenvolvimento , Regulação para Cima
10.
Artigo em Inglês | MEDLINE | ID: mdl-9185275

RESUMO

The cells of Burkholderia pseudomallei, B. cepacia and Pseudomonas aeruginosa grown on agar plates were stained with fluorescently-labeled insulin. The former two species were stained positively indicating insulin binding but P. aeruginosa was not. Insulin exposure reduced phospholipase C and acid phosphatase activities of B. pseudomallei but did not affect those enzymatic activities of B. cepacia in the employed experimental conditions. It is suggested that B. pseudomallei have insulin receptors which may be associated with a signal transfer system involving phospholipase and protein tyrosine phosphatase.


Assuntos
Infecções por Burkholderia/enzimologia , Burkholderia cepacia/enzimologia , Burkholderia pseudomallei/enzimologia , Complicações do Diabetes , Insulina/farmacologia , Fosfatase Ácida/metabolismo , Infecções por Burkholderia/complicações , Infecções por Burkholderia/microbiologia , Burkholderia cepacia/patogenicidade , Burkholderia pseudomallei/patogenicidade , Diabetes Mellitus/microbiologia , Humanos , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Fosfolipases Tipo C/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA