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1.
Chinese Journal of Virology ; (6): 44-50, 2013.
Article Dans Chinois | WPRIM | ID: wpr-339976

Résumé

Vpr, an auxiliary protein of HIV-1(Human immunodeficiency virus type 1), exerts important functions to promote viral replication and AIDS progression. In this study, we performed a yeast two-hybrid screening assay using human cDNA library to further investigate the molecular mechanism of various functions of Vpr RelB, a key protein in NF-kappaB signaling pathway, was identified as a Vpr interaction protein by co-immunoprecipitation. Further investigations indicated that RelB not only promoted the Vpr-mediated activation of NF-kappaB reporter gene, but also enhanced the transactivation of HIV LTR. Moreover, the results showed that RelB promoted Vpr-induced cell cycle G2/M arrest. Collectively, these results indicated that RelB might interact with Vpr and regulate its transcriptional activation and cell cycle arrest.


Sujets)
Humains , Points de contrôle du cycle cellulaire , Division cellulaire , Phase G2 , Répétition terminale longue du VIH , Cellules HeLa , Facteur de transcription NF-kappa B , Génétique , Facteur de transcription RelB , Physiologie , Activation de la transcription , Produits du gène vpr du virus de l'immunodéficience humaine , Physiologie
2.
Chinese Medical Journal ; (24): 2874-2879, 2009.
Article Dans Anglais | WPRIM | ID: wpr-266023

Résumé

<p><b>BACKGROUND</b>The CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.</p><p><b>METHODS</b>A SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.</p><p><b>RESULTS</b>One SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.</p><p><b>CONCLUSIONS</b>We conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.</p>


Sujets)
Animaux , Humains , Chimère , Gènes env , VIH-1 (Virus de l'Immunodéficience Humaine de type 1) , Génétique , Physiologie , Macaca mulatta , Provirus , Génétique , Récepteurs CCR5 , Physiologie , Virus de l'immunodéficience simienne , Génétique , Physiologie
3.
Chinese Journal of Experimental and Clinical Virology ; (6): 143-145, 2003.
Article Dans Chinois | WPRIM | ID: wpr-250520

Résumé

<p><b>OBJECTIVE</b>Chimeric human/bovine immunodeficiency virus (HBIV) cDNA was constructed by replacing HIV tat and LTR with bovine immunodeficiency virus (BIV) tat and LTR to study the activity of BIV tat and LTR in the chimerae.</p><p><b>METHODS</b>The target fragments of BIV tat, LTR and HIV gag, pol, env were respectively amplified by using PCR and sequentially inserted into pBluescript SK(+) vector. The chimeric clone was transfected into human MT4 cells. The transcript and gene expression of the HBIV chimeric virus were detected by using RT-PCR and a reverse transcriptase assay, respectively.</p><p><b>RESULTS</b>BIV tat mRNA and HIV gag mRNA were detected. The reverse transcriptase activity of the chimeric virus was analyzed in the fluctuation curve.</p><p><b>CONCLUSIONS</b>In chimeric HBIV cDNA transfected MT?4 cells, BIV tat and HIV gag were transcripted. The reverse transcriptase of the chimeric virus had biological activity. These data suggest that in MT4 cells, BIV LTR had promoter activity and BIV tat had the function of transactivation in the chimeric virus. The study of the chimeric virus with infectivity is in progress.</p>


Sujets)
Animaux , Bovins , Humains , Vaccins contre le SIDA , Clonage moléculaire , ADN complémentaire , Génétique , ADN viral , Génétique , Gènes gag , Génétique , Gènes pol , Génétique , Gènes tat , Génétique , VIH-1 (Virus de l'Immunodéficience Humaine de type 1) , Génétique , Virus de l'immunodéficience bovine , Génétique , Protéines de fusion recombinantes , Génétique , Métabolisme , Transcription génétique , Activation de la transcription , Transfection , Réplication virale
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