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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2010.tde-06042010-110031
Document
Author
Full name
Bruna Felicio Milazzotto Maldonado Porchia
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Ferreira, Luis Carlos de Souza (President)
Pereira, Carlos Augusto
Rodrigues, Mauricio Martins
Title in Portuguese
Construção e análise das propriedades profiláticas e terapêuticas de uma vacina contra tumores associados ao HPV-16.
Keywords in Portuguese
Glicoproteínas D
HPV-16
Neoplasias
Purificação de proteínas
Vacinas
Abstract in Portuguese
O desenvolvimento de vacinas contra o vírus do papiloma humano (HPV) representa uma importante alternativa para o controle da infecção sexualmente transmissível e do câncer cervical. Neste trabalho exploramos uma estratégia vacinal inédita contra tumores induzidos pelo HPV-16 empregando a proteína E7 obtida após fusão genética com a glicoproteína D do vírus herpes tipo 1, uma proteína com propriedades adjuvantes para linfócitos T. A proteína recombinante gDE7 foi expressa em bactérias e em células de inseto e purificada por cromatografia de afinidade. A proteína gerada em bactérias foi administrada nas formas insolúvel e solúvel como vacina em camundongos. A gDE7 insolúvel conferiu 80% de proteção profilática para o crescimento tumoral, a gDE7 que manteve a forma solúvel após refolding protegeu 100% dos animais nas mesmas condições. Já a proteção terapêutica alcançou 30% dos animais. Além disso, verificamos o potencial neutralizante dos soros gerados frente ao HSV-1. As condições de obtenção das proteínas expressas em células de inseto também foram estabelecidas.
Title in English
Construction and analysis of the prophylactic and therapeutic proprieties of a vaccine against HPV-16 associated tumors.
Keywords in English
Glycoproteins D
HPV-16
Neoplasms
Protein purification
Vaccines
Abstract in English
The development of vaccines against human papillomavirus (HPV) represents an important alternative to control the sexually transmitted infection and cervical cancer. In this paper we explored a novel vaccine strategy against tumors induced by HPV-16 using the E7 protein obtained after genetic fusion to the glycoprotein D of herpes virus type 1, a protein with adjuvant properties for T lymphocytes. The recombinant protein gDE7 was expressed in bacteria and in insect cells and purified by affinity chromatography. The protein generated in bacteria was administered in soluble and insoluble forms as a vaccine in mice. The insoluble gDE7 gave 80% prophylactic protection for tumor growth, the gDE7 that kept the soluble form after refolding protected 100% of the animals under the same conditions. The therapeutic protection reached 30% of the animals. We also verified the neutralizing potential of sera generated against the HSV-1. The conditions for obtaining the proteins expressed in insect cells were also established.
 
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Publishing Date
2010-04-06
 
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