Chim3 confers survival advantage to CD4+ T cells upon HIV-1 infection by preventing HIV-1 DNA integration and HIV-1–induced G2 cell-cycle delay

Porcellini, Simona; Gubinelli, Francesco; Alberici, Luca; Piovani, Bianca; Rizzardi, Gian-Paolo; Bovolenta, Chiara

doi:10.1182/blood-2009-09-243030

Cited by 14 publications

(19 citation statements)

References 26 publications

(55 reference statements)

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“…The F12-Vif derivative Chim3-Vif is a chimera Vif generated by replacing the domain (aa 126-170) of wild-type Vif with that of F12-Vif. Lentiviral transduction of Chim3-Vif protected CD4 + T cells from HIV-1 infection by preventing HIV-1 DNA integration and HIV-1-induced G2 cell cycle delay (47,48). These data also indicate that modification of the cell cycle is important for HIV-1 replication and might be a target for an original therapeutic strategy.…”

Section: Discussionmentioning

confidence: 67%

HIV-1 viral infectivity factor interacts with TP53 to induce G2 cell cycle arrest and positively regulate viral replication

Izumi

Io²,

Matsui³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Viral infectivity factor, an accessory protein encoded in the HIV-1 genome, induces G2 cell cycle arrest; however, the biological significance and mechanism(s) remain totally unclear. Here we demonstrate that the TP53 pathway is involved in Vif-mediated G2 cell cycle arrest. Vif enhances the stability and transcriptional activity of TP53 by blocking the MDM2-mediated ubiquitination and nuclear export of TP53. Furthermore, Vif causes G2 cell cycle arrest in a TP53-dependent manner. HXB2 Vif lacks these activities toward TP53 and cannot induce G2 cell cycle arrest. Using mutagenesis, we demonstrate that the critical residues for this function are located in the Nterminal region of Vif. Finally, we construct a mutant NL4-3 virus with an NL4-3/HXB2 chimeric Vif defective for the ability to induce cell cycle arrest and show that the mutant virus replicates less effectively than the wild-type NL4-3 virus in T cells expressing TP53. These data imply that Vif induces G2 cell cycle arrest through functional interaction with the TP53/MDM2 axis and that the G2 cell cycle arrest induced by Vif has a positive effect on HIV-1 replication. This report demonstrates the molecular mechanisms and the biological significance of Vif-mediated G2 cell cycle arrest for HIV-1 infection.AIDS | NL4-3 | HXB2 | APOBEC3G | infectivity

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Section: Discussionmentioning

confidence: 67%

HIV-1 viral infectivity factor interacts with TP53 to induce G2 cell cycle arrest and positively regulate viral replication

Izumi

Io²,

Matsui³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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“…1a, schemes 5 and 6). The second-generation PDNChim3 TV was previously described (Porcellini et al, , 2010 (Fig. 1a, scheme 7).…”

Section: Plasmidsmentioning

confidence: 99%

“…The viral pellets were resuspended in Iscove's modified Dulbecco's medium supplemented with 10% fetal bovine serum and frozen at -70°C (Strang et al, 2004). Titer was calculated on different cell lines by one or two cycles of spinoculation at 1,240 g for 45 min in the presence of polybrene 8 lg/ml (Sigma-Aldrich); CD34 + HSCs were transduced for 24 hr on retronectin-coated plates (Takara Bio) as previously described (Porcellini et al, , 2010. Transduction efficiency was monitored by flow cytometry analysis (FACS Calibur BD Bioscience) of eGFP (SIN-eGFP) or DLNFGR expression (PDN-Chim3), as previously described (Porcellini et al, , 2010.…”

Section: Baculovirus Production and Baculo-gpr Infection Of Hek-293t mentioning

confidence: 99%

“…+ cells and neonatal leukocytes were prestimulated for 48 hr in a cytokine cocktail and maintained in the same medium during transduction and cultivation as previously described (Porcellini et al, , 2010.…”

Section: Cellsmentioning

confidence: 99%

“…Furthermore, significant progress has been recently made using stem-cell-based gene and cell therapy approaches to treat HIV-1 infection (Kitchen et al, 2011). In this context, we have previously demonstrated that the therapeutic gene Chim3, an HIV-1 Vif-dominant negative factor, when delivered to target cells by means of Tat-conditional LV, inhibits HIV-1 replication and confers selective growth advantage to transduced cells (Porcellini et al, , 2010. Chim3-based anti-HIV gene therapy is therefore a promising therapeutic intervention, which requires an adequate clinical-grade manufacturing to be further tested in preclinical animal models or in patients.…”

mentioning

confidence: 99%

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RD2-MolPack-Chim3,a Packaging Cell Line for Stable Production of Lentiviral Vectors for Anti-HIV Gene Therapy

Stornaiuolo

Piovani

Bossi

et al. 2013

Human Gene Therapy Methods

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Over the last two decades, several attempts to generate packaging cells for lentiviral vectors (LV) have been made. Despite different technologies, no packaging clone is currently employed in clinical trials. We developed a new strategy for LV stable production based on the HEK-293T progenitor cells; the sequential insertion of the viral genes by integrating vectors; the constitutive expression of the viral components; and the RD114-TR envelope pseudotyping. We generated the intermediate clone PK-7 expressing constitutively gag/pol and rev genes and, by adding tat and rd114-tr genes, the stable packaging cell line RD2-MolPack, which can produce LV carrying any transfer vector (TV). Finally, we obtained the RD2-MolPack-Chim3 producer clone by transducing RD2-MolPack cells with the TV expressing the anti-HIV transgene Chim3. Remarkably, RD114-TR pseudovirions have much higher potency when produced by stable compared with transient technology. Most importantly, comparable transduction efficiency in hematopoietic stem cells (HSC) is obtained with 2-logs less physical particles respect to VSV-G pseudovirions produced by transient transfection. Altogether, RD2-MolPack technology should be considered a valid option for large-scale production of LV to be used in gene therapy protocols employing HSC, resulting in the possibility of downsizing the manufacturing scale by about 10-fold in respect to transient technology.

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Problems and Prospects of Gene Therapy Against HIV

Schneider¹,

Wagner²,

Davydova³

et al. 2014

Pharm Chem J

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Chim3 confers survival advantage to CD4+ T cells upon HIV-1 infection by preventing HIV-1 DNA integration and HIV-1–induced G2 cell-cycle delay

Cited by 14 publications

References 26 publications

HIV-1 viral infectivity factor interacts with TP53 to induce G2 cell cycle arrest and positively regulate viral replication

HIV-1 viral infectivity factor interacts with TP53 to induce G2 cell cycle arrest and positively regulate viral replication

RD2-MolPack-Chim3,a Packaging Cell Line for Stable Production of Lentiviral Vectors for Anti-HIV Gene Therapy

Problems and Prospects of Gene Therapy Against HIV

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