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Master's Dissertation
DOI
10.11606/D.87.2010.tde-26082010-163625
Document
Author
Full name
Paulo Silva Wolf
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Gebara, Vera Cristina Bugelli Cainelli (President)
Quintilio, Wagner
Silva, Wilmar Dias da
Title in Portuguese
Clonagem, expressão e avaliação da imunogenicidade e do potencial adjuvante induzidos pela proteína "heat-shock"  Cpn60 da Bordetella pertussis.
Keywords in Portuguese
Adjuvantes imunológicos
Bactérias aeróbicas Gram-negativas
Clonagem molecular
Coqueluche
Proteínas do choque térmico
Vacinas
Abstract in Portuguese
A proteína Cpn60 faz parte de um grupo de proteínas altamente conservadas que estão envolvidas em funções celulares essenciais. camundongos BALB\c foram imunizados com 5 ou 10 µg da proteína recombinante (Cpn60r) sozinha ou adicionada à vacina DTP sem hidróxido de alumínio (NADTP). A vacina DTP do Instituto Butantan (DTP) foi usada como controle. Foi avaliada a produção de citocinas por células esplênicas após reestímulo in vitro com a Cpn60r. Os animais foram desafiados após o protocolo de imunização. A Cpn60r sozinha ou misturada à vacina NADTP foi capaz de induzir níveis de anticorpos contra pertussis mais altos do que os induzidos pela DTP. Os níveis de IgG1 e IgG2a foram similares para todos os grupos. Pôde-se observar a produção de de IL-6 e IFN-γ nos grupos imunizados com Cpn60r. Os grupos imunizados com Cpn60r+NADTP apresentaram um índice de proteção entre 60 e 80% contra o desafio pela bactéria virulenta, semelhante ao grupo imunizado com DTP. A proteína Cpn60r é bastante promissora não somente como imunógeno, mas também como adjuvante.
Title in English
Molecular cloning, expression and evaluation of immunogenicity and adjuvant potential induced from the heat-shock protein Cpn60 from Bordetella pertussis.
Keywords in English
Gram-negative aerobic bacteria
Heat-shock proteins
Immunologic adjuvants
Molecular cloning
Vaccines
Whooping cough
Abstract in English
The Cpn60 protein is a member of a group of higly conserved proteins linked to essencial cell functions. The Cpn60 was cloned, expressed and its immune response has been evaluated. BALB\c mice were immunized with 5 or 10 µg of the recombinant protein (Cpn60r) alone or mixed with DPT vaccine without aluminum hidroxyde (NADPT). The DPT vaccine from Instituto Butantan was used as control. We evaluated the cytokines production by spleen cells after they have been reestimulated in vitro with Cpn60r. The animals were challenged after the immunization protocol. The Cpn60r alone or mixed with NADPT vaccine was able to induce higher antibodies levels than DPT. IgG1 and IgG2a levels were similar in all groups. We could detect levels of IL-6 and IFN-γ on groups immunized with Cpn60r. The groups immunized with Cpn60r+NADTP showed a 60 and 80% protection rate against the challenge with the live bacteria, similar to the group immunized with DPT. These results show the immune response of the recombinant protein that could be included in immunization protocols for pertussis.
 
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Publishing Date
2010-09-16
 
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