The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion
An increased plasma concentration of von Willebrand factor (vWF) is detected in individuals with many infectious diseases and is accepted as a marker of endothelium activation and prothrombotic condition. To determine whether ExoU, a Pseudomonas aeruginosa cytotoxin with proinflammatory activity, en...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2012-09-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600004&lng=en&tlng=en |
| _version_ | 1827879818323034112 |
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| author | Carla Freitas Maria-Cristina Assis Alessandra Mattos Saliba Veronica Maria Morandi Camila Castro Figueiredo Mirian Pereira Maria-Cristina Plotkowski |
| author_facet | Carla Freitas Maria-Cristina Assis Alessandra Mattos Saliba Veronica Maria Morandi Camila Castro Figueiredo Mirian Pereira Maria-Cristina Plotkowski |
| author_sort | Carla Freitas |
| collection | DOAJ |
| description | An increased plasma concentration of von Willebrand factor (vWF) is detected in individuals with many infectious diseases and is accepted as a marker of endothelium activation and prothrombotic condition. To determine whether ExoU, a Pseudomonas aeruginosa cytotoxin with proinflammatory activity, enhances the release of vWF, microvascular endothelial cells were infected with the ExoU-producing PA103 P. aeruginosa strain or an exoU-deficient mutant. Significantly increased vWF concentrations were detected in conditioned medium and subendothelial extracellular matrix from cultures infected with the wild-type bacteria, as determined by enzyme-linked immunoassays. PA103-infected cells also released higher concentrations of procoagulant microparticles containing increased amounts of membrane-associated vWF, as determined by flow cytometric analyses of cell culture supernatants. Both flow cytometry and confocal microscopy showed that increased amounts of vWF were associated with cytoplasmic membranes from cells infected with the ExoU-producing bacteria. PA103-infected cultures exposed to platelet suspensions exhibited increased percentages of cells with platelet adhesion. Because no modulation of the vWF mRNA levels was detected by reverse transcription-polymerase chain reaction assays in PA103-infected cells, ExoU is likely to have induced the release of vWF from cytoplasmic stores rather than vWF gene transcription. Such release is likely to modify the thromboresistance of microvascular endothelial cells. |
| first_indexed | 2024-03-12T18:11:43Z |
| format | Article |
| id | doaj.art-d2658967a744485caabb3209ef33864e |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T18:11:43Z |
| publishDate | 2012-09-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-d2658967a744485caabb3209ef33864e2023-08-02T09:15:23ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-80602012-09-01107672873410.1590/S0074-02762012000600004S0074-02762012000600004The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesionCarla Freitas0Maria-Cristina Assis1Alessandra Mattos Saliba2Veronica Maria Morandi3Camila Castro Figueiredo4Mirian Pereira5Maria-Cristina Plotkowski6Universidade do Estado do Rio de JaneiroCentro Universitário Estadual da Zona OesteUniversidade do Estado do Rio de JaneiroUniversidade do Estado do Rio de JaneiroUniversidade do Estado do Rio de JaneiroFundação Oswaldo CruzUniversidade do Estado do Rio de JaneiroAn increased plasma concentration of von Willebrand factor (vWF) is detected in individuals with many infectious diseases and is accepted as a marker of endothelium activation and prothrombotic condition. To determine whether ExoU, a Pseudomonas aeruginosa cytotoxin with proinflammatory activity, enhances the release of vWF, microvascular endothelial cells were infected with the ExoU-producing PA103 P. aeruginosa strain or an exoU-deficient mutant. Significantly increased vWF concentrations were detected in conditioned medium and subendothelial extracellular matrix from cultures infected with the wild-type bacteria, as determined by enzyme-linked immunoassays. PA103-infected cells also released higher concentrations of procoagulant microparticles containing increased amounts of membrane-associated vWF, as determined by flow cytometric analyses of cell culture supernatants. Both flow cytometry and confocal microscopy showed that increased amounts of vWF were associated with cytoplasmic membranes from cells infected with the ExoU-producing bacteria. PA103-infected cultures exposed to platelet suspensions exhibited increased percentages of cells with platelet adhesion. Because no modulation of the vWF mRNA levels was detected by reverse transcription-polymerase chain reaction assays in PA103-infected cells, ExoU is likely to have induced the release of vWF from cytoplasmic stores rather than vWF gene transcription. Such release is likely to modify the thromboresistance of microvascular endothelial cells.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600004&lng=en&tlng=enPseudomonas aeruginosavon Willebrand factorprothrombotic activitymicroparticles |
| spellingShingle | Carla Freitas Maria-Cristina Assis Alessandra Mattos Saliba Veronica Maria Morandi Camila Castro Figueiredo Mirian Pereira Maria-Cristina Plotkowski The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion Memorias do Instituto Oswaldo Cruz Pseudomonas aeruginosa von Willebrand factor prothrombotic activity microparticles |
| title | The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion |
| title_full | The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion |
| title_fullStr | The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion |
| title_full_unstemmed | The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion |
| title_short | The infection of microvascular endothelial cells with ExoU-producing Pseudomonas aeruginosa triggers the release of von Willebrand factor and platelet adhesion |
| title_sort | infection of microvascular endothelial cells with exou producing pseudomonas aeruginosa triggers the release of von willebrand factor and platelet adhesion |
| topic | Pseudomonas aeruginosa von Willebrand factor prothrombotic activity microparticles |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600004&lng=en&tlng=en |
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