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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2010.tde-02032011-173909
Document
Author
Full name
Erika Nakajima
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Martins, Elizabeth Angelica Leme (President)
Almeida, Maria Elisabete Sbrogio de
Aniz, Patricia Antonia Estima Abreu de
Piazza, Roxane Maria Fontes
Vasconcellos, Silvio Arruda
Title in Portuguese
Investigação de proteínas candidatas vacinais contra leptospirose. Apresentação de antígenos na forma de proteínas recombinantes purificadas ou como vacinas vivas em salmonelas atenuadas.
Keywords in Portuguese
Antígenos de bactérias
Genética bacteriana
Leptospirose
Recombinação genética
Vacinas
Abstract in Portuguese
A leptospirose é uma doença endêmica causada por Leptospiras. O genoma da Leptospira interrogans sorovar Copenhageni foi analisado para seleção de potenciais antígenos vacinais. Oito genes foram selecionados e clonados para expressão e purificação dos antígenos. A salmonela SL3261 foi usada como carregadora dos genes de leptospira em vetor pAEsox para expressão das proteínas in vivo. As salmonelas recombinantes induziram resposta imune quando administradas em camundongos por via intraperitoneal. Hamsters foram imunizados com as salmonelas, observando-se que a SLLIC10191 induziu proteção parcial no desafio com L. interrogans sorovar Pomona. Vetores híbridos foram construídos para expressão simultânea de dois antígenos em salmonelas in vivo. Observamos indução de anticorpos específicos, porém, os ensaios de desafio não foram conclusivos. Vários parâmetros do desafio com sorovar Copenhageni foram estudados, como contagem das bactérias e ajuste de dose, variação de virulência por passagens em cultivo e interferência da idade dos animais.
Title in English
Investigation of proteins vaccine candidates against leptospirosis. Antigens presentation as purified recombinant proteins or as live vaccines by attenuated salmonelas.
Keywords in English
Bacterial antigens
Bacterial genetics
Genetic recombination
Leptospirosis
Vaccines
Abstract in English
Leptospirosis is an endemic disease caused by Leptospira. The genome of Leptospira interrogans serovar Copenhageni was analyzed for screening potential vaccine antigens. Eight genes were selected and cloned for expression and purification. Salmonella SL3261 was used as carrier of the genes of leptospira in pAEsox vector for in vivo proteins expression of proteins in vivo. Recombinant Salmonella induced immune response when administered intraperitoneally in mice intraperitoneally. Hamsters were immunized with salmonella, resulting we observed that the SLLIC10191 induced partial protection against on challenge with L. interrogans serovar Pomona. Hybrid vectors were constructed for expression of two antigens simultaneously by salmonella in vivo. We observed induction of specific antibodies, however, the challenge tests were not conclusive. Several parameters of the challenge assay with serovar Copenhageni were studied, such as the counting of bacteria count and dose adjustment, changes in virulence by passages in culture and interference backgroundof from the age of animals.
 
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Publishing Date
2011-08-11
 
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