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Master's Dissertation
Document
Author
Full name
Lazara Elena Santiesteban Lores
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Tanizaki, Martha Massako (President)
Gonçalves, Viviane Maimoni
Lemos, Ana Paula Silva de
Title in Portuguese
Síntese, caracterização e avaliação imunológica de conjugados do sorotipo 6B de Streptococcus pneumoniae à proteína A de superfície pneumocócica (PspA).
Keywords in Portuguese
Streptococcus pneumoniae
Conjugação
PspA
Abstract in Portuguese
As vacinas conjugadas contra Streptococcus pneumoniae mostram-se eficazes na redução da doença pneumocócica, no entanto depois de sua introdução tem se verificado a substituição de sorotipos. Para evitar estes problemas o emprego de proteínas da própria bactéria como carreadoras tem sido usado. Neste trabalho a PspA foi conjugada ao sorotipo 6B de S. pneumoniae por dois métodos de conjugação. Inicialmente a PspA clado 1 e 3 foram purificadas, recuperando as proteínas com mais de 90% de pureza. A conjugação intermediada pelo agente ativador cloreto de 4-(4,6-dimetoxi-1,3,5-triazin-2-il-)-4-metilmorfolino (DMT-MM) permitiu rendimentos de Ps entre 20 e 30%, no entanto este tipo de conjugado não foi imunogênico resultando na não indução de anticorpos anti-PspA. Por outro lado a conjugação por aminação redutiva possibilitou obter rendimentos de Ps entre 50 e 60% e os conjugados induziram elevados títulos de anticorpos anti-PspA em camundongos Balb/c, que foram capazes de promover a fagocitose da bactéria; no entanto, a resposta de anticorpos anti-Ps 6B foi baixa.
Title in English
Synthesis, characterization and immunological evaluation of conjugates from Streptococcus pneumoniae serotype 6B and Pneumococcal Surface Protein A (PspA).
Keywords in English
Streptococcus pneumoniae
Conjugation
PspA
Abstract in English
Conjugate vaccines against Streptococcus pneumonaie have had an important public health benefit; nevertheless after its introduction it has been observed a serotype replacement. To solve this problems conjugate vaccines using pneumococcal surface proteins as carriers has been studied. In this work, Pneumococal surface protein A (PspA) was employed as a carrier protein, conjugated to serotype 6B. Initially PspA clade 1 and PspA clade 3 were purified; both proteins were recovered with more than 90% purity. The conjugation mediated by the activating agent 4- (4,6-dimethoxy-1,3,5-triazin-2-yl -) - 4-methylmorpholine chloride(DMT-MM) allowed Ps yields between 20 and 30%, however this conjugation chemistry had a negative impact on the immunogenicity of the protein. On the other hand conjugation by reductive amination led to Ps yields between 50 and 60% and induced high anti-PspA antibodies titers in Balb /c mice that were able to promote phagocytosis of the bacterium; however, polysaccharide 6B induced low antibody titers.
 
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Publishing Date
2017-06-19
 
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