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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2015.tde-04092015-143806
Document
Author
Full name
Daniella Cristina Ventini Monteiro
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Pereira, Carlos Augusto (President)
Augusto, Elisabeth de Fatima Pires
Azzoni, Adriano Rodrigues
Núñez, Eutimio Gustavo Fernández
Soares, Carlos Roberto Jorge
Title in Portuguese
Purificação e imunogenicidade da glicoproteína do vírus da raiva (RVGP) expressa pelos sistemas células S2 e Semliki Forest Virus.
Keywords in Portuguese
Células de inseto S2
Glicoproteína do vírus da raiva
Vacina contra raiva
Vírus Semliki Forest
Abstract in Portuguese
O desenvolvimento de vacinas contra a raiva tão eficientes quanto as atuais ainda é considerado importante para a profilaxia dessa doença devido ao alto número de mortes por ano no mundo. Este trabalho mostra dois sistemas para a expressão recombinante do principal antígeno da raiva, a glicoproteína viral (RVGP): células Schneider 2 de Drosophila melanogaster estavelmente transfectadas (S2 rRVGP) e vírus Semliki Forest (SFV) carregando o RNA da RVGP (SFVRVGP). Ensaios de purificação por cromatografia de afinidade, da rRVGP de S2 rRVGP produzida em biorreator, demonstraram resultados promissores para o isolamento de monômeros da rRVGP. Para os estudos de imunogenicidade, camundongos foram vacinados com rRVGP de S2 rRVGP e SFV-RVGP. Dosagem de anticorpos anti-glicoproteína do vírus da raiva, neutralizantes, IgG1 e IgG2a, e citocinas demonstraram que o SFV-RVGP induziu predominantemente uma resposta imune do tipo celular e que os dois vetores foram capazes de expressar uma rRVGP imunogênica, apresentando um potencial uso clínico (veterinário e humano).
Title in English
Purification and immunogenicity of the rabies virus glycoprotein (RVGP) expressed by S2 cells and Semliki Forest Virus systems.
Keywords in English
Semliki Forest Virus
Rabies vaccine
Rabies virus glycoprotein
S2 insect cells
Abstract in English
The development of new and equally efficient rabies vaccines is still considered important to the prophylaxis of the disease, which is responsible for many deaths per year worldwide. This work used two systems for recombinant expression of the major rabies antigen, the viral glycoprotein (RVGP): stably transfected Drosophila melanogaster Schneider 2 cells (S2 rRVGP) and Semliki Forest Virus (SFV) carrying the RNA of RVGP (SFV-RVGP). Purification assays of the rRVGP by metal ion affinity chromatography, from S2 rRVGP cells produced in bioreactor, showed promising results for rRVGP monomers isolation. Analysis of antibodies anti-rabies virus glycoprotein, neutralizing, IgG1 and IgG2a, and citokines showed that the SFV-RVGP induced predominantly a cellular immune response, and both vectors were capable of expressing an immunogenic rRVGP, what reinforces potential clinical applications (veterinary or human).
 
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Publishing Date
2015-09-04
 
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