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Doctoral Thesis
DOI
10.11606/T.87.2011.tde-09022012-090823
Document
Author
Full name
Bogar Omar Araujo Montoya
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Leite, Luciana Cezar de Cerqueira (President)
Daffre, Sirlei
Marco, Ricardo De
Santelli, Glaucia Maria Machado
Winter, Carlos Eduardo
Title in Portuguese
Investigação da Carboxipeptidase, Esfingomielinase e Fosfatase Alcalina de Schistosoma mansoni como potenciais antígenos.
Keywords in Portuguese
Schistosoma mansoni
Alcalina
Carboxipeptidase
Esfingomielinase
Proteínas recombinantes
Vacinas
Abstract in Portuguese
A esquistossomose representa um grande problema de saúde pública em regiões tropicais, negligenciado pelas empresas farmacêuticas. Três genes foram selecionados a partir do transcriptoma do S. mansoni para serem caracterizados molecularmente e testados como vacinas. O gene da Carboxipeptidase apresentou altos níveis de transcrição no estágio de cercária e uma variável expressão protéica nos estágios intra-hospedeiros. A proteína recombinante renaturada apresentou baixa atividade. O gene da Esfingomielinase apresentou alto nível transcricional em ovos, sendo expresso em todos os estágios exceto em esquistossômulos de 7 dias e fêmeas. O gene da Fosfatase Alcalina apresentou elevada transcrição em cercária, mas a expressão da proteína se eleva somente no estágio de esquistossômulo. A imunização de camundongos com cada uma das três proteínas recombinantes expressas em E. coli não levou à redução da carga parasitária após desafio. Estes resultados demonstram que diversas características de um antígeno são necessárias para que este seja um bom candidato vacinal.
Title in English
Investigation of Carboxypeptidase, Sphingomyelinase and Alkaline Phosphatase from Schistosoma mansoni as potential vaccine candidates.
Keywords in English
Schistosoma mansoni
Alkaline
Carboxypeptidase
Recombinant proteins
Sphingomyelinase
Vaccines
Abstract in English
Schistosomiasis is a major public health problem in tropical areas and neglected by most of the pharmaceutical companies. Three genes were selected from S. mansoni transcriptome to be molecularly characterized and tested as vaccines. The Carboxypeptidase gene showed high transcription levels in cercariae stage and a variable protein expression in intra-host stages. The refolded recombinant protein showed limited activity. The Sphingomyelinase gene showed the highest level of transcriptional activity in the egg stage and the protein is expressed in all stages but 7-day old schistosomula and females. The Alkaline Phosphatase gene showed a high transcriptional activity in cercariae stage with most of the mRNA being translated into protein as soon as it penetrates its definitive host. Immunization of mice with each of the three proteins did not show reduction in worm burden recovery after challenge. These results demonstrate that several characteristics of an antigen are important for it to be considered a good vaccine candidate.
 
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Publishing Date
2012-03-01
 
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