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Doctoral Thesis
DOI
10.11606/T.87.2014.tde-08122014-143251
Document
Author
Full name
Marcos Alexandre Nobre Lemos
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Jorge, Soraia Attie Calil (President)
Domingos, Marta de Oliveira
Durigon, Edison Luiz
Pinto, Jorge Luiz Freire
Santos, Milene Cristina Menezes dos
Title in Portuguese
Expressão gênica da proteína não estrutural 3 do vírus da hepatite C empregando pseudopartículas virais.
Keywords in Portuguese
Expressão heteróloga
HCV
HCVpp
Hepatite C
NS3
Proteína não estrutural 3 do HCV
Pseudopartículas virais
SFV
Abstract in Portuguese
A hepatite viral causada pelo vírus da hepatite C (HCV) é um problema à saúde mundial e afeta cerca de 170 milhões de pessoas. O caso crônico da doença resulta em cirrose hepática e a maioria dos pacientes tratados não desenvolve uma resposta imune satisfatória. A proteína não estrutural 3 (NS3) pode estimular uma resposta celular que auxilia a resposta nos infectados. Nosso trabalho desenvolveu duas pseudopartículas virais que carregam um material genético para a protease da NS3 do HCV. Um dos sistemas, a HCVpp é constituída por proteínas do vírus da leucemia murina e outras do HCV. E o outro sistema, a partícula viral é baseada no Semliki Forest Virus (SFV). As células HEK293T e BHK-21 foram transfectadas para a formação das pseudopartículas HCVpp-NS3p1a e SFV-NS3p1a, respectivamente. Essas partículas foram quantificadas pela presença do material genético da NS3 por qRT-PCR, atingindo uma produção aproximada de 4x105 partículas HCVpp-NS3p1a/mL e 2,5x107 partículas SFV-NS3p1a/mL. Essas partículas foram utilizadas para infecção de células HuH-7.0 e BHK-21.
Title in English
Gene expression of the nonstructural protein 3 of hepatitis C virus using viral pseudoparticles.
Keywords in English
HCV
HCV nonstructural protein 3
HCVpp
Hepatitis C
Heterologous expression
NS3
SFV
Viral pseudoparticles
Abstract in English
Viral hepatitis caused by the hepatitis C virus (HCV) is a global health problem, affecting about 170 million people. The chronic case of the disease results in liver cirrhosis and most patients do not develop a satisfactory immune response. The nonstructural protein 3 (NS3) can stimulate a cellular response that helps answer the infected. Our work has developed two viral pseudoparticles who carry a genetic material for the HCV NS3 protease. One of the systems is constituted by the HCVpp proteins of murine leukemia virus and other HCV. The other system, the viral particle is based on the Semliki Forest Virus (SFV). The HEK293T and BHK-21 cells were transfected for forming the pseudoparticles HCVpp-NS3p1a and SFV-NS3p1a, respectively. These particles were quantified by the presence of genetic material of NS3 by qRT-PCR, reaching a production of about 4x105 HCVpp-NS3p1a particles/mL and 2,5 x107 SFV-NS3p1a particles /mL. These particles were used for infection of Huh-7.0 cells and BHK-21.
 
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Release Date
2016-12-09
Publishing Date
2014-12-10
 
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