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Doctoral Thesis
DOI
10.11606/T.42.2008.tde-25112008-121908
Document
Author
Full name
Luis Ernesto Farinha Arcieri
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Ventura, Armando Morais (President)
Arruda Neto, Eurico de
Durigon, Edison Luiz
Felippe, Julia Maria Martins de Souza
Ferreira, Luis Carlos de Souza
Title in Portuguese
Desenvolvimento e avaliação de ferramentas de imunização baseadas na região globular da fibra adenoviral modificada com o domínio C4 da glicoproteína gp120 do HIV.
Keywords in Portuguese
Adenovírus
Baculoviridae
HIV
Imunização
Proteínas recombinantes
Vacinas
Abstract in Portuguese
A glicoproteína gp120 do HIV apresenta domínios conservados, dentre os quais está o C4. Este domínio está envolvido no reconhecimento da molécula CD4 na superfície das células alvo, e anticorpos dirigidos contra ele neutralizam o vírus. Em trabalho anterior, este domínio foi introduzido dentro da região globular da proteína fibra do adenovírus. Como a fibra adenoviral é estimulante do sistema imune, decidimos testar a capacidade de indução de anticorpos anti-C4 por essa proteína modificada. Assim, foram construídos vetores plasmídicos e adenovirais portando o gene da região globular da fibra adenoviral contendo o domínio C4, que usados na imunização de camundongos induziram anticorpos capazes de reconhecer a proteína gp120. Também construímos um vetor baculoviral expressando essa proteína híbrida, que purificada por HPLC, foi utilizada para imunizar camundongos que também produziram anticorpos capazes de reconhecer a proteína gp120. Nossos dados sugerem que a região globular da fibra adenoviral é uma boa plataforma para a exposição de epítopos de imunização.
Title in English
Development and evaluation of immunization tools based on the adenovirus fiber knob modified with the C4 domain of HIV gp120 glycoprotein.
Keywords in English
Baculoviridae
Adenovirus
HIV
Immunization
Recombinant proteins
Vaccines
Abstract in English
HIV glycoprotein gp120 has conserved domains, one of them being the C4 domain. This region is involved in the recognition of the CD4 marker in target cells and antibodies that recognize this domain can block HIV infection. Previously, the C4 domain was introduced in the adenovirus fiber knob. As the adenovirus fiber stimulates de immune system, we decided to test the production of anti-C4 antibodies by this hybrid protein. We constructed plasmid and adenovirus vectors carrying the fiber knob modified with the C4 domain. Immunization of mice with these vectors showed the production of specific antibodies that recognized de gp120 glycoprotein. Also, we constructed a baculovirus vector expressing the hybrid protein, which was purified by HPLC. Mice immunized with this protein also produced antibodies capable of recognizing gp120. Our data suggest that the fiber knob is a good carrier protein for epitope immunization.
 
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Publishing Date
2009-05-13
 
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