A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFMA |
Texto Completo: | https://tedebc.ufma.br/jspui/handle/tede/tede/2904 |
Resumo: | Some Escherichia coli strains are pathogens responsible for a variety of diseases, and albeit using distinct mechanisms of pathogenesis, have in common the production of Pic, or protein involved in colonization. We have recently shown that Pic mediates immune evasion by the direct cleavage of complement molecules, significantly reducing complement activation by all three pathways. The aim of this study was to investigate the action of Pic in a murine model of sepsis. Six to eight-week-old female Swiss mice were intraperitoneally inoculated with Pic-producing E. coli (F5), F5Δpic mutant or apyrogenic water (control). Animal survival was monitored for 5 days (n= 6 animals/group) and a subset of mice (n= 6 animals/group) was euthanized after 12 h for sample acquire from blood, peritoneal lavage and organs. The parameters evaluated were cellularity from blood, peritoneal cavity and organs; inflammatory mediators production; quantity of colony forming units (CFU's); immunophenotyping of cells from spleen and peritoneum; and histopathology of organs. Intraperitoneal inoculation of Pic-producing bacteria induced 100% death within 24 h, different from the other groups where no death was observed. The CFU count of bacteria in the organs was significantly higher in F5 than other groups. Besides, only F5 was viable on bloodstream. Hematological analysis showed a decrease of total leukocytes, mainly lymphocytes. NO and cytokines (IFN-γ, TNF-α, IL-6, IL-12, IL-10 and MCP-1) were detected in serum, as well as on peritoneal lavage (excepting IL-12) of F5 group in significantly higher levels than other groups. Immunophenotyping of spleen and peritoneum cells revealed that there was a low expression of co-receptors on lymphocytes and macrophages in F5 group, indicating that the activation and function of these were compromised. Therefore, these results evidenced that Pic represents an important virulence factor, allowing the survival of the bacterium on bloodstream and several organs, as well as inducing a high production of pro-inflammatory mediators by the host, leading to a sepsis and death. |
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ABREU JUNIOR, Afonso Gomes965677243-15http://lattes.cnpq.br/4394413983541820ABREU JUNIOR, Afonso Gomes965677243-15http://lattes.cnpq.br/4394413983541820MONTEIRO NETO, Valériohttp://lattes.cnpq.br/5476942147520772SILVA, Luís Cláudio Nascimento dahttp://lattes.cnpq.br/6016850820500623MACIEL, Marcia Cristina Gonçalveshttp://lattes.cnpq.br/0645092224285117035290183-76http://lattes.cnpq.br/0174848340560198DUTRA , Itaynara Lobato2019-11-13T17:08:23Z2018-04-27DUTRA, Itaynara Lobato. A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino. 2018. 80 f. Dissertação (Programa de Pós-Graduação em Ciências da Saúde/CCBS) - Universidade Federal do Maranhão, São Luís, 2018.https://tedebc.ufma.br/jspui/handle/tede/tede/2904Some Escherichia coli strains are pathogens responsible for a variety of diseases, and albeit using distinct mechanisms of pathogenesis, have in common the production of Pic, or protein involved in colonization. We have recently shown that Pic mediates immune evasion by the direct cleavage of complement molecules, significantly reducing complement activation by all three pathways. The aim of this study was to investigate the action of Pic in a murine model of sepsis. Six to eight-week-old female Swiss mice were intraperitoneally inoculated with Pic-producing E. coli (F5), F5Δpic mutant or apyrogenic water (control). Animal survival was monitored for 5 days (n= 6 animals/group) and a subset of mice (n= 6 animals/group) was euthanized after 12 h for sample acquire from blood, peritoneal lavage and organs. The parameters evaluated were cellularity from blood, peritoneal cavity and organs; inflammatory mediators production; quantity of colony forming units (CFU's); immunophenotyping of cells from spleen and peritoneum; and histopathology of organs. Intraperitoneal inoculation of Pic-producing bacteria induced 100% death within 24 h, different from the other groups where no death was observed. The CFU count of bacteria in the organs was significantly higher in F5 than other groups. Besides, only F5 was viable on bloodstream. Hematological analysis showed a decrease of total leukocytes, mainly lymphocytes. NO and cytokines (IFN-γ, TNF-α, IL-6, IL-12, IL-10 and MCP-1) were detected in serum, as well as on peritoneal lavage (excepting IL-12) of F5 group in significantly higher levels than other groups. Immunophenotyping of spleen and peritoneum cells revealed that there was a low expression of co-receptors on lymphocytes and macrophages in F5 group, indicating that the activation and function of these were compromised. Therefore, these results evidenced that Pic represents an important virulence factor, allowing the survival of the bacterium on bloodstream and several organs, as well as inducing a high production of pro-inflammatory mediators by the host, leading to a sepsis and death.Algumas linhagens de Escherichia coli são patógenos responsáveis por uma variedade doenças e, embora usando mecanismos distintos de patogênese, têm em comum a produção da Proteína envolvida na colonização (Pic). Recentemente nosso grupo mostrou que Pic medeia a evasão do sistema imune pela clivagem direta de moléculas do sistema complemento, reduzindo a ativação deste em todas as três vias. O objetivo deste estudo foi investigar a ação de Pic em um modelo murino de sepse. Camundongos fêmeas da linhagem Swiss entre 6-8 semanas receberam um inoculo intraperitoneal de suspenções bacterianas contendo E. coli produtora de Pic (F5), o mutante F5Δpic, ou uma injeção de água apirogênica. A sobrevida dos animais foi acompanhada durante 5 dias (n=6 animais/grupo). A outra parte dos animais (n=6 animais/grupo) foi eutanasiada 12 h após a infecção para obtenção das amostras de sangue, lavado peritoneal e órgãos. Os parâmetros avaliados foram: a quantidade de células do sangue, cavidade peritoneal e órgãos; produção de mediadores inflamatórios; determinação das unidades formadoras de colônias (UFCs); imunofenotipagem de células do baço e peritônio; e histopatologia dos órgãos. A inoculação intraperitoneal de bactérias produtoras de Pic induziu a morte de 100% dos animais em até 24h, diferente dos demais grupos entre os quais não foi observada mortalidade. A quantidade de UFCs dos órgãos foi mais alta no grupo F5 que nos outros grupos. Além disso, apenas F5 permaneceu viável na corrente sanguínea. O hemograma mostrou uma redução no número total de leucócitos, especialmente de linfócitos, nos animais do grupo F5 quando comparado aos demais grupos. Óxido nítrico, citocinas (IFN-γ, TNF-α, IL-6, IL-12, IL-10) e a quimiocina MCP-1 foram detectadas no soro, bem como no lavado peritoneal (exceto IL-12) do grupo F5 de modo mais acentuado que nos demais grupos. A imunofenotipagem das células do baço e peritônio evidenciou que houve uma baixa expressão de co-receptores em linfócitos e macrófagos no grupo F5, indicando que a ativação e a função destes foi comprometida. Esses resultados demonstram que Pic representa um importante fator de virulência, permitindo a sobrevivência da bactéria na corrente sanguínea e vários órgãos, assim como induzindo a alta produção de mediadores pró-inflamatórios pelo hospedeiro, levando à sepse e morte.Submitted by Sheila MONTEIRO (sheila.monteiro@ufma.br) on 2019-11-13T17:08:23Z No. of bitstreams: 1 ITAYNARA-DUTRA.pdf: 2589366 bytes, checksum: da5acd1c71f0560ba04d1e49fd8ecb79 (MD5)Made available in DSpace on 2019-11-13T17:08:23Z (GMT). No. of bitstreams: 1 ITAYNARA-DUTRA.pdf: 2589366 bytes, checksum: da5acd1c71f0560ba04d1e49fd8ecb79 (MD5) Previous issue date: 2018-04-27Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão - FAPEMAapplication/pdfporUniversidade Federal do MaranhãoPROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS DA SAÚDE/CCBSUFMABrasilDEPARTAMENTO DE BIOLOGIA/CCBSEscherichia coliPicSerinaproteaseSepseEscherichia coliPicSerineproteaseSepsisCiências da SaúdeA proteína pic produzida por Escherichia coli induz sepse letal em modelo murinoEscherichia coli-produced pic protein induces lethal sepsis in a murine modelinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFMAinstname:Universidade Federal do Maranhão (UFMA)instacron:UFMAORIGINALITAYNARA-DUTRA.pdfITAYNARA-DUTRA.pdfapplication/pdf2589366http://tedebc.ufma.br:8080/bitstream/tede/2904/2/ITAYNARA-DUTRA.pdfda5acd1c71f0560ba04d1e49fd8ecb79MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82255http://tedebc.ufma.br:8080/bitstream/tede/2904/1/license.txt97eeade1fce43278e63fe063657f8083MD51tede/29042019-11-13 14:08:23.294oai:tede2: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Biblioteca Digital de Teses e Dissertaçõeshttps://tedebc.ufma.br/jspui/PUBhttp://tedebc.ufma.br:8080/oai/requestrepositorio@ufma.br||repositorio@ufma.bropendoar:21312019-11-13T17:08:23Biblioteca Digital de Teses e Dissertações da UFMA - Universidade Federal do Maranhão (UFMA)false |
dc.title.por.fl_str_mv |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
dc.title.alternative.eng.fl_str_mv |
Escherichia coli-produced pic protein induces lethal sepsis in a murine model |
title |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
spellingShingle |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino DUTRA , Itaynara Lobato Escherichia coli Pic Serinaprotease Sepse Escherichia coli Pic Serineprotease Sepsis Ciências da Saúde |
title_short |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
title_full |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
title_fullStr |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
title_full_unstemmed |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
title_sort |
A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino |
author |
DUTRA , Itaynara Lobato |
author_facet |
DUTRA , Itaynara Lobato |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
ABREU JUNIOR, Afonso Gomes |
dc.contributor.advisor1ID.fl_str_mv |
965677243-15 |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4394413983541820 |
dc.contributor.referee1.fl_str_mv |
ABREU JUNIOR, Afonso Gomes |
dc.contributor.referee1ID.fl_str_mv |
965677243-15 |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/4394413983541820 |
dc.contributor.referee2.fl_str_mv |
MONTEIRO NETO, Valério |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/5476942147520772 |
dc.contributor.referee3.fl_str_mv |
SILVA, Luís Cláudio Nascimento da |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/6016850820500623 |
dc.contributor.referee4.fl_str_mv |
MACIEL, Marcia Cristina Gonçalves |
dc.contributor.referee4Lattes.fl_str_mv |
http://lattes.cnpq.br/0645092224285117 |
dc.contributor.authorID.fl_str_mv |
035290183-76 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/0174848340560198 |
dc.contributor.author.fl_str_mv |
DUTRA , Itaynara Lobato |
contributor_str_mv |
ABREU JUNIOR, Afonso Gomes ABREU JUNIOR, Afonso Gomes MONTEIRO NETO, Valério SILVA, Luís Cláudio Nascimento da MACIEL, Marcia Cristina Gonçalves |
dc.subject.por.fl_str_mv |
Escherichia coli Pic Serinaprotease Sepse Escherichia coli Pic Serineprotease Sepsis |
topic |
Escherichia coli Pic Serinaprotease Sepse Escherichia coli Pic Serineprotease Sepsis Ciências da Saúde |
dc.subject.cnpq.fl_str_mv |
Ciências da Saúde |
description |
Some Escherichia coli strains are pathogens responsible for a variety of diseases, and albeit using distinct mechanisms of pathogenesis, have in common the production of Pic, or protein involved in colonization. We have recently shown that Pic mediates immune evasion by the direct cleavage of complement molecules, significantly reducing complement activation by all three pathways. The aim of this study was to investigate the action of Pic in a murine model of sepsis. Six to eight-week-old female Swiss mice were intraperitoneally inoculated with Pic-producing E. coli (F5), F5Δpic mutant or apyrogenic water (control). Animal survival was monitored for 5 days (n= 6 animals/group) and a subset of mice (n= 6 animals/group) was euthanized after 12 h for sample acquire from blood, peritoneal lavage and organs. The parameters evaluated were cellularity from blood, peritoneal cavity and organs; inflammatory mediators production; quantity of colony forming units (CFU's); immunophenotyping of cells from spleen and peritoneum; and histopathology of organs. Intraperitoneal inoculation of Pic-producing bacteria induced 100% death within 24 h, different from the other groups where no death was observed. The CFU count of bacteria in the organs was significantly higher in F5 than other groups. Besides, only F5 was viable on bloodstream. Hematological analysis showed a decrease of total leukocytes, mainly lymphocytes. NO and cytokines (IFN-γ, TNF-α, IL-6, IL-12, IL-10 and MCP-1) were detected in serum, as well as on peritoneal lavage (excepting IL-12) of F5 group in significantly higher levels than other groups. Immunophenotyping of spleen and peritoneum cells revealed that there was a low expression of co-receptors on lymphocytes and macrophages in F5 group, indicating that the activation and function of these were compromised. Therefore, these results evidenced that Pic represents an important virulence factor, allowing the survival of the bacterium on bloodstream and several organs, as well as inducing a high production of pro-inflammatory mediators by the host, leading to a sepsis and death. |
publishDate |
2018 |
dc.date.issued.fl_str_mv |
2018-04-27 |
dc.date.accessioned.fl_str_mv |
2019-11-13T17:08:23Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
DUTRA, Itaynara Lobato. A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino. 2018. 80 f. Dissertação (Programa de Pós-Graduação em Ciências da Saúde/CCBS) - Universidade Federal do Maranhão, São Luís, 2018. |
dc.identifier.uri.fl_str_mv |
https://tedebc.ufma.br/jspui/handle/tede/tede/2904 |
identifier_str_mv |
DUTRA, Itaynara Lobato. A proteína pic produzida por Escherichia coli induz sepse letal em modelo murino. 2018. 80 f. Dissertação (Programa de Pós-Graduação em Ciências da Saúde/CCBS) - Universidade Federal do Maranhão, São Luís, 2018. |
url |
https://tedebc.ufma.br/jspui/handle/tede/tede/2904 |
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por |
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por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal do Maranhão |
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PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS DA SAÚDE/CCBS |
dc.publisher.initials.fl_str_mv |
UFMA |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
DEPARTAMENTO DE BIOLOGIA/CCBS |
publisher.none.fl_str_mv |
Universidade Federal do Maranhão |
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