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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2014.tde-24022015-124003
Document
Author
Full name
Renata Damasio de Souza
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Ferreira, Luis Carlos de Souza (President)
Ho, Paulo Lee
Huenuman, Nilton Erbet Lincopan
Rosa, Daniela Santoro
Stephano, Marco Antonio
Title in Portuguese
Esporos de Bacillus subtilis como adjuvante vacinal.
Keywords in Portuguese
Bacillus subtilis
Adjuvante
Esporos
Gag-p24
Métodos de purificação
Abstract in Portuguese
Esporos de Bacillus subtilis apresentam propriedades adjuvantes, sendo capazes de aumentar a resposta humoral após a sua coadministração com antígenos misturados ou adsorvidos à sua superfície. Mas, para isso, é necessária a produção de esporos altamente purificados e com rendimentos elevados. Neste trabalho, realizamos com sucesso uma análise quantitativa das condições de esporulação e dos métodos de purificação, o que melhorou a reprodutibilidade do processo e a obtenção de amostras com elevado grau de pureza e rendimento. Avaliamos também as propriedades imunomodulatórias destes esporos, utilizando como antígeno modelo a proteína recombinante Gag-p24 do HIV-1. A coadministração, mas não a adsorção à superfície do esporo, aumentou a imunogenicidade do antígeno sem induzir efeitos deletérios após a administração parenteral em camundongos BALB/c e C57BL/6. Além de promoveram a ativação das APCs, os esporos interagem com receptores relacionados à imunidade inata, devido à ausência do efeito adjuvante em camundongos nocautes para TLR2. Esses resultados abrem perspectivas interessantes para a utilização de esporos como adjuvantes vacinais.
Title in English
Bacillus subtilis spores as a vaccine adjuvante.
Keywords in English
Bacillus subtilis
Adjuvant
Gag-p24
Purification methods
Spores
Abstract in English
Bacillus subtilis spores have been shown to behave as vaccine adjuvants, promoting the increase of antibody responses after co-administration with antigens either admixed or adsorbed on the spore surface. Nonetheless, such specialized application requires highly purified spore preparations at high yields. In this work, we successfully performed a systematic quantitative analysis of sporulation conditions and spore purification methods, which improved the reproducibility of the process and the obtainment of samples with high purity and yield. Afterwards, we further evaluated the immune modulatory properties of these spores using a recombinant HIV-1 Gag-p24 protein as a model antigen. The co-administration, but not adsorption to the spore surface, enhanced the immunogenicity of that target antigen, without inducing deleterious effects, after subcutaneous administration to BALB/c and C57BL/6 mice. Besides promoting activation of antigen presenting cells, spores interact with receptors related to innate immunity, due to the absence of the adjuvant effect on TLR2 knockout mice. These results open interesting perspectives for the use of B. subtilis spores as vaccine adjuvants.
 
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Publishing Date
2015-02-24
 
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