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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

ModulaÃÃo imunolÃgica preventiva aplicada ao controle de infecÃÃes bacterianas por salmonella typhimurium pelo uso de lectinas / Preventive immune modulation applied to the control of bacterial infections by Salmonella typhimurium by the use of lectins

Ayrles Fernanda BrandÃo da Silva 22 April 2014 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / DoenÃas infecciosas constituem a principal causa de morte em todo o mundo. Dentre estas, as causadas por infecÃÃes bacterianas se destacam devido à diversidade de bactÃrias e à sua capacidade evolutiva e adaptativa. Respostas inflamatÃrias inadequadas estÃo associadas a essas infecÃÃes e em detrimento de avanÃos cientÃficos, o diagnÃstico precoce e o tratamento de quadros de infecÃÃes bacterianas sistÃmicas permanecem um desafio à saÃde pÃblica. Neste contexto, este estudo avaliou a capacidade da lectina de Canavalia brasiliensis (ConBr) e Cratylia argentea (CFL) em modular a inflamaÃÃo sistÃmica causada por Salmonella enterica sorotipo Typhimurium. As lectinas foram purificadas por cromatografia de afinidade em coluna de Sephadex G-50 e ensaios de avaliaÃÃo toxicolÃgica e seguranÃa farmacolÃgica foram realizados. Ambas as lectinas nÃo demonstraram toxicidade atà a mÃxima dose de 2000 mg/kg. EntÃo, camundongos foram inoculados, por via intraperitoneal, com as lectinas (10 mg/kg), 72, 48 e 24 horas antes de serem infectados com a Salmonella (107 UFC/mL). Foi observada uma sobrevida de 100% e 90% dos animais tratados com a ConBr e CFL, respectivamente, avaliados no sÃtimo dia. Este protocolo reduziu significativamente o nÃmero de bactÃrias no sangue, fluÃdo peritoneal e nos principais ÃrgÃos da infecÃÃo (baÃo e fÃgado). Foi observado ainda reversÃo da falÃncia na migraÃÃo de neutrÃfilos, leucopenia, bem como, reduÃÃo nos nÃveis de citocinas (TNF-α, IL-1 e IL-10) e Ãxido nÃtrico no soro. Foi tambÃm verificado um aumento na concentraÃÃo de Ãxido nÃtrico no fluido peritoneal e do nÃmero de plaquetas no sangue. AnÃlises in vitro mostraram que ambas as lectinas nÃo apresentam atividade bactericida, reduzem o tempo de ativaÃÃo do sistema complemento e aumentam o tempo de coagulaÃÃo intrÃnseca e extrÃnseca. As lectinas nÃo apresentaram efeito curativo quando administradas apÃs a infecÃÃo e nÃo demonstraram efeito preventivo quando inoculadas por via endovenosa ou oral. AtravÃs da anÃlise proteÃmica, proteÃnas relacionadas à infecÃÃo e à aÃÃo moduladora das lectinas foram identificadas. Assim, as lectinas possivelmente atuam sobre os macrÃfagos/monÃcitos, controlando sua ativaÃÃo, e desta forma, atenuando o processo inflamatÃrio e protegendo os animais do choque sÃptico. Estes resultados representam uma interessante perspectiva para o controle de infecÃÃes, com uso independente ou associado a antibiÃticos de aÃÃo direta sobre microrganismos / Infectious diseases are a leading cause of death throughout the world. Among these, bacterial infections stand out due to the enormous diversity of bacteria and their evolutionary and adaptive abilities. Inappropriate inflammatory responses have been associated with these infections and despite contemporaneous scientific approaches, timely diagnosis and proper treatment of systemic bacterial infection are still a challenge for public health. Thus, this study evaluated the ability of Canavalia brasiliensis (ConBr) and Cratylia argentea (CFL) lectin in modulating systemic inflammation caused by Salmonella enterica serovar Typhimurium. The lectins were purified by affinity chromatography on Sephadex G50. Toxicology and the pharmacological security of lectins was evaluated. Both lectins exhibited no toxicity up to 2000 mg/kg. Mice were inoculated intraperitoneally with lectins (10 mg/kg) within 72, 48 and 24 h before infection with Salmonella (107 CFU/mL). Animals treated with ConBr and CFL, respectively, showed 100% and 90% survival, seven days after infection. This protocol significantly reduced the bacteria in blood, peritoneal fluid and in main organs such as liver and spleen. It was also observed a reversal of the neutrophil migration failure, leukopenia, as well as reduction of cytokine levels (TNF- α, IL- 1 and IL -10) and nitric oxide on the serum. In addition, elevated peritoneal fluid concentration of nitric oxide and increase in the number of platelets were found. In vitro analyzes identified that both lectins did not exhibit antibacterial activity, can decrease the time of activation of complement system and increase intrinsic and extrinsic clotting time. Lectins had no curative effect when administered after infection and none protective effect when inoculated either intravenously or orally. Through proteomic analysis, proteins related to infection and to modulating action of lectins were identified. According to our results, we found that lectins can act on macrophages/monocytes, controlling their activation and thereby influencing the inflammatory process, protecting the animals from septic shock. These results represent an interesting approach for the control of infections, using lectins independently or associated with antibiotics whose action is directly on microorganisms.

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