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Doctoral Thesis
DOI
10.11606/T.87.2014.tde-19022015-093553
Document
Author
Full name
Ivana Barros de Campos
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Gonçalves, Viviane Maimoni (President)
Gomez, José Gregorio Cabrera
Guglielmi, Luiza Guilherme
Piccoli, Rosane Aparecida Moniz
Tonso, Aldo
Title in Portuguese
Otimização do processo de produção e caracterização da vacina celular contra Streptococcus pneumoniae.
Keywords in Portuguese
Avaliação imunológica de vacinas
Cultivo contínuo com reciclo de células
Ensaio opsonofagocítico
Pneumococo
Vacina celular pneumocócica
Abstract in Portuguese
S. pneumoniae é um patógeno de grande impacto em saúde pública e vacinas comerciais têm cobertura limitada e alto custo. Como alternativa, desenvolveu-se uma vacina celular de baixo custo, cuja produção envolve apenas a separação das bactérias do caldo e sua inativação. Neste trabalho, foram avaliados processos descontínuo, descontínuo alimentado e contínuo com reciclo de células, cuja produção de biomassa foi 3 vezes maior que a do descontínuo. Vacinas obtidas nos 3 processos foram utilizadas em ensaios de imunização de camundongos e induziram níveis similares de IgG e IL-17A. Anticorpos ligaram-se e induziram a deposição de moléculas do sistema complemento sobre a superfície do pneumococo. Ademais, induziram fagocitose de diferentes cepas encapsuladas da bactéria. Camundongos imunizados foram protegidos contra sepse após aspiração da cepa virulenta WU2. Portanto, o processo contínuo com reciclo permitiu a obtenção de maior número de doses sem alterar a qualidade da vacina e o ensaio opsonofagocítico poderia ser utilizado como potencial correlato de proteção.
Title in English
Optimization of the production process and characterization of Streptococcus pneumoniae whole cell vaccine.
Keywords in English
Continuous fermentation with cell recycle
Immunological evaluation of the vaccine
Opsonophagocytic assay
Pneumococcal whole cell vaccine
Pneumococcus
Abstract in English
S. pneumoniae is a pathogen of great impact on public health and commercially available vaccines have limited coverage and high cost. As alternative, a low-cost whole cell vaccine was developed, whose production involves only the cell separation and inactivation. In this work, we evaluated batch, fed-batch and continuous cultivation with cell recycle. The biomass production was 3-fold higher in continuous process than batch. Vaccines obtained from these 3 processes were used to immunize mice and all vaccines induced comparable levels of IgG and IL-17A. Antibodies were able to bind and induce deposition of complement onto pneumococcal surface, besides to induce phagocytosis of several encapsulated pneumococcal strains in opsonophagocytic assays. Immunized mice were protected from fatal aspiration-sepsis using the virulent pneumococcal strain WU2. Therefore, the continuous process with cell recycle yielded a higher number of doses without altering the quality of the vaccine and opsonophagocytic assay could be used as a potential correlate of protection.
 
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Release Date
2017-02-19
Publishing Date
2015-02-20
 
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