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1.
J Virol ; 82(7): 3574-83, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18234788

RESUMO

The characterization of internal ribosome entry sites (IRESs) in virtually all lentiviruses prompted us to investigate the mechanism used by the feline immunodeficiency virus (FIV) to produce viral proteins. Various in vitro translation assays with mono- and bicistronic constructs revealed that translation of the FIV genomic RNA occurred both by a cap-dependent mechanism and by weak internal entry of the ribosomes. This weak IRES activity was confirmed in feline cells expressing bicistronic RNAs containing the FIV 5' untranslated region (UTR). Surprisingly, infection of feline cells with FIV, but not human immunodeficiency virus type 1, resulted in a great increase in FIV translation. Moreover, a change in the cellular physiological condition provoked by heat stress resulted in the specific stimulation of expression driven by the FIV 5' UTR while cap-dependent initiation was severely repressed. These results reveal the presence of a "dormant" IRES that becomes activated by viral infection and cellular stress.


Assuntos
Vírus da Imunodeficiência Felina/fisiologia , Biossíntese de Proteínas , RNA Viral/genética , Ribossomos/metabolismo , Proteínas Virais/biossíntese , Regiões 5' não Traduzidas , Animais , Sítios de Ligação , Gatos , Linhagem Celular
2.
J Virol ; 80(18): 8856-68, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16940498

RESUMO

Determining which antigen must be included in AIDS vaccines to confer maximum protection is of utmost importance. In primate models, vaccines consisting of or including accessory viral proteins have yielded conflicting results. We investigated the protective potential of the accessory protein ORF-A of feline immunodeficiency virus (FIV) in cats. All three immunization strategies used (protein alone in alum adjuvant, DNA alone, or DNA prime-protein boost) clearly generated detectable immune responses. Upon challenge with ex vivo homologous FIV, ORF-A-immunized cats showed distinct enhancement of acute-phase infection relative to mock-immunized animals given alum or empty vector DNA. This effect was tentatively attributed to increased expression of the FIV receptor CD134 that was observed in the immunized cats. However, at subsequent sampling points that were continued for up to 10 months postchallenge, the average plasma viral loads of the ORF-A-immunized animals were slightly but consistently reduced relative to those of the control animals. In addition, CD4(+) T lymphocytes in the circulation system declined more slowly in immunized animals than in control animals. These findings support the contention that immunization with lentiviral accessory proteins can improve the host's ability to control virus replication and slow down disease progression but also draw attention to the fact that even simple immunogens that eventually contribute to protective activity can transiently exacerbate subsequent lentiviral infections.


Assuntos
Vacinas contra a AIDS/química , Síndrome da Imunodeficiência Adquirida/prevenção & controle , DNA Viral/química , Vírus da Imunodeficiência Felina/metabolismo , Proteínas Virais/química , Sequência de Aminoácidos , Animais , Linfócitos T CD4-Positivos/metabolismo , Células CHO , Gatos , Cricetinae , Humanos , Dados de Sequência Molecular , Receptores OX40 , Receptores do Fator de Necrose Tumoral/biossíntese , Carga Viral , Proteínas Virais/genética
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