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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2009.tde-23102009-113942
Document
Author
Full name
Camila Mathias dos Santos
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Ferreira, Luis Carlos de Souza (President)
Ho, Paulo Lee
Piazza, Roxane Maria Fontes
Title in Portuguese
Avaliação das propriedades imunomoduladoras de toxinas termo-lábeis do tipo II produzidas por Escherichia coli enterotoxigênica (ETEC) administradas por via transcutânea.
Keywords in Portuguese
Escherichia coli
Adjuvantes imunológicos
Adjuvantes vacinais
Imunização intradérmica
Imunização transcutânea
Microbiologia
Ovalbumina
Toxinas termo-lábeis
Vacinas
Abstract in Portuguese
Toxinas termo-lábeis expressas por Escherichia coli enterotoxigênica (LT-I, LT-IIa e LT-IIb) são adjuvantes sistêmicos e de mucosa. Essas proteínas apresentam reduzida identidade (<14%) em suas subunidades B e ligação a receptores distintos, o que pode resultar em propriedades biológicas diferenciais. O objetivo do trabalho foi avaliar a resposta imune induzida pela toxina LT-IIa e seu pentâmero B (LT-IIaB) administradas pelas vias transcutânea e intradérmica. Inicialmente as proteínas foram expressas em E. coli, purificadas e avaliadas quanto à funcionalidade in vitro. Numa segunda etapa, as propriedades imunomoduladoras de LT-IIa e LT-IIaB, imunogenicidade e atividade adjuvante, foram avaliadas em modelo murino. A resposta imune (humoral e celular) induzida contra ovalbumina (OVA), aplicada como antígeno, foi determinada. Os resultados demonstram que as atividades adjuvantes induzidas por LT-IIa e LT-IIaB variam de acordo com a via de inoculação e que as holotoxinas LT-I e LT-IIa induzem graus de inflamação e ativação de resposta imune distintos em camundongos.
Title in English
Evaluation of the immunomodulatory properties of type II heat-labile toxins produced by enterotoxigenic Escherichia coli (ETEC) administered by transcutaneous route.
Keywords in English
Escherichia coli
Heat labile toxins
Immunological adjuvants
Intradermic immunization
Microbiology
Ovalbumin
Transcutaneous immuniozation
Vaccine adjuvants
Vaccines
Abstract in English
Heat-labile toxins expressed by enterotoxigenic Escherichia coli (LT-I, LT-IIa and LT-IIb) are potent systemic and mucosal adjuvants. These proteins have low identity (<14%) in their B subunits and bind to different receptors, which may result in differential biological properties. The objective of this work was to evaluate the immune response induced by LT-IIa toxin and its pentameric B subunit (LT-IIaB) delivered by transcutaneous and intradermic routes. Initially the proteins of interest were expressed in recombinant E. coli strains, purified and tested for functionality in vitro. In a second moment, the immunomodulatory properties of LT-IIa and LT-IIaB, immunogenicity and adjuvant activity, were evaluated in mouse model. The humoral and cellular immune responses induced against ovalbumin (OVA) used as antigen were determined. The results show that the adjuvant activity of LT-IIa and its B pentamer depends on the route of inoculation and that LT-I and LT-IIa differ both on induction of inflammation and activation of immune responses in mice.
 
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Publishing Date
2009-12-04
 
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