RESUMO
INTRODUCTION: Persistent infection with GBV-C (GB Virus C), a non-pathogenic virus related to hepatitis C virus (HCV), prolongs survival in HIV infection. Two GBV-C proteins, NS5A and E2, have been shown previously to inhibit HIV replication in vitro. We investigated whether the GBV-C NS3 serine protease affects HIV replication. RESULTS: GBV-C NS3 protease expressed in a human CD4+ T lymphocyte cell line significantly inhibited HIV replication. Addition of NS4A or NS4A/4B coding sequence to GBV-C NS3 increased the effect on HIV replication. Inhibition of HIV replication was dose-dependent and was not mediated by increased cell toxicity. Mutation of the NS3 catalytic serine to alanine resulted in loss of both HIV inhibition and protease activity. GBV-C NS3 expression did not measurably decrease CD4 or CXCR4 expression. CONCLUSION: GBV-C NS3 serine protease significantly inhibited HIV replication without decreasing HIV receptor expression. The requirement for an intact catalytic serine at the active site indicates that inhibition was mediated by proteolytic cleavage of an unidentified target(s).
Assuntos
Linfócitos T CD4-Positivos/metabolismo , Vírus GB C/enzimologia , HIV-1/efeitos dos fármacos , Receptores de HIV/genética , Proteínas não Estruturais Virais/genética , Proteínas não Estruturais Virais/farmacologia , Replicação Viral/efeitos dos fármacos , Sequência de Aminoácidos , Fármacos Anti-HIV/metabolismo , Fármacos Anti-HIV/farmacologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/virologia , Linhagem Celular , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Vírus GB C/genética , Expressão Gênica/efeitos dos fármacos , Genoma Viral/genética , HIV-1/fisiologia , Humanos , Células Jurkat , Dados de Sequência Molecular , RNA Helicases/genética , RNA Helicases/metabolismo , RNA Helicases/farmacologia , RNA Helicases/fisiologia , Receptores de HIV/metabolismo , Homologia de Sequência de Aminoácidos , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Serina Endopeptidases/farmacologia , Serina Endopeptidases/fisiologia , Transfecção , Proteínas não Estruturais Virais/metabolismo , Proteínas não Estruturais Virais/fisiologia , Replicação Viral/genéticaRESUMO
GB virus C (GBV-C) infection is common in humans and may persist for decades, although most infected persons clear the virus and subsequently develop antibodies to the envelope glycoprotein. GBV-C replicates in peripheral blood mononuclear cells (PBMCs) and CD4(+) T lymphocytes in vitro, and depletion of CD4(+) T lymphocytes has been proposed as the reason for clearance of GBV-C among persons positive for human immunodeficiency virus. We identified GBV-C RNA in purified CD4(+) and CD8(+) T lymphocytes and CD19(+) B lymphocytes removed ex vivo from infected donors and found that GBV-C replicated in vitro in these PBMC subsets, suggesting that GBV-C is a panlymphotropic virus.
Assuntos
Linfócitos B/virologia , Linfócitos T CD4-Positivos/virologia , Linfócitos T CD8-Positivos/virologia , Vírus GB C/fisiologia , Replicação Viral , Antígenos CD19/metabolismo , Células Cultivadas , Infecções por Flaviviridae/complicações , Infecções por Flaviviridae/virologia , Vírus GB C/genética , Vírus GB C/isolamento & purificação , Infecções por HIV/complicações , Infecções por HIV/virologia , Hepatite Viral Humana/complicações , Hepatite Viral Humana/virologia , Humanos , RNA Viral/sangueRESUMO
GB virus C (GBV-C) is a nonpathogenic member of the Flaviviridae family most closely related to hepatitis C virus (HCV). Infection is common in healthy and immunocompromised people and may persist for years. GBV-C infection is associated with improved survival, improved AIDS-free survival, higher CD4(+) T-cell counts, and lower HIV viral loads in HIV-infected people compared with people infected with HIV but not GBV-C. The mechanism of this effect is not yet clear, but GBV-C has been shown to inhibit HIV replication in vitro through increased synthesis and secretion of anti-HIV b-chemokines MIP-1a, MIP-1b, RANTES, SDF-1, and SDF-2 and downregulation of CCR5 receptor expression. GBV-C also inhibits apoptosis of its host cell, similar to HCV. GBV-C E2 protein in serum has also been associated with prolonged survival in HIV infection; recent evidence indicates that GBV-C E2 protein may neutralize HIV infection in vitro.
