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Master's Dissertation
DOI
10.11606/D.87.2013.tde-08052013-111313
Document
Author
Full name
Luis Guilherme Virgilio Fernandes
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Nascimento, Ana Lucia Tabet Oller do (President)
Elias Junior, Waldir Pereira
Ferreira, Rita de Cassia Cafe
Title in Portuguese
Caracterização e avaliação imunológica de três proteínas de superfície de Leptospira interrogans obtidas em Escherichia coli
Keywords in Portuguese
Escherichia coli
Camundongos
Genômica
Leptospirose
Proteínas recombinates
Abstract in Portuguese
A leptospirose é uma zoonose causada por bactérias patogênicas do gênero Leptospira. O sequenciamento do genoma completo de L. interrogans sorovar Copenhageni tem permitido a obtenção e caracterização de proteínas potencialmente envolvidas na patogênese desta bactéria, como lipoproteínas e proteínas de membrana externa. Neste trabalho, foram estudados três genes, OmpL1, LIC10731 e LIC10645, dos quais o gene OmpL1 foi o mais frequente em diferentes espécies de Leptospira. As proteínas recombinantes foram purificadas por cromatografia de afinidade ao metal. As três proteínas recombinantes promoveram resposta humoral e celular após imunização em camundongos. Ensaios de adesão mostraram que as proteínas se ligam à laminina e plasminogênio, e adicionalmente a proteína OmpL1 se liga ao fibrinogênio e fibronectina plasmática. A proteína OmpL1 foi bastante reativa com soro de pacientes de leptospirose. Os resultados sugerem que as proteínas referentes aos genes estudados podem desempenhar um papel na patogênese da bactéria.
Title in English
Characterization and immunological evaluation of three surface proteins of Leptospira interrogans obtained in Escherichia coli.
Keywords in English
Escherichia coli
Genomics
Leptospirosis
Mice
Recombinant proteins
Abstract in English
Leptospirosis is a zoonosis caused by pathogenic bacteria of genus Leptospira. Annotation of the genome sequences of L. interrogans serovar Copenhageni allows the identification and characterization of proteins potentially involved in the pathogenesis of this bacterium, such as lipoproteins and outer-membrane proteins. The present study characterized three genes, OmpL1, LIC10731 and LIC10645, and one of them, OmpL1, was the most frequent among different species of Leptospira. The recombinant proteins were purified by metal-chelating chromatography. All three recombinant proteins promoted humoral and cellular response after immunization in mice. Binding assays showed that all proteins interact to laminin and plasminogen, and additionally protein OmpL1 binds to fibrinogen and plasma fibronectin. OmpL1 was highly reactive with positive-leptospirosis human sera. The results suggest that the proteins encoded by these genes may play a role in the bacterium pathogenesis.
 
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Publishing Date
2013-06-13
 
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