New adenovirus vectors for protein production and gene transfer

Massie, Bernard; Mosser, Dick D.; Koutroumanis, Maria; Vitté-Mony, I; Lamoureux, Linda; Couture, France; Paquet, Luc; Guilbault, Claire; Dionne, Julie; Chahla, Dounia; Jolicœur, Pierre; Langelier, Yves

doi:10.1007/978-94-011-4786-6_7

Cited by 27 publications

(64 citation statements)

References 26 publications

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“…BMAdE1 220-8, 293A, and 293rtTA [15,25,26] were propagated in antibiotic-free Dulbecco minimal essential medium (DMEM) with high glucose concentration supplemented with 10% tetracycline-free fetal bovine serum (FBS) (Clontech Laboratories Inc., Palo Alto, BVDV NS3-induced apoptosis 215 USA). 293A and BMAdE1 cells were used for AdV generation and amplification [15,26] whereas 293rtTA cells were used to assess the expression of the protein of interest and to titrate the adenovirus stocks by flow cytometry monitoring of GFP expression [26]. A549 cells (derived from human lung carcinoma tissues) genetically transformed to express the tetracycline transactivation factor (tTA) (A549tTA) [25] were used in cell apoptosis experiments conducted with the AdV.…”

Section: Cells and Virusesmentioning

confidence: 99%

“…This transfer vector enables the gene of interest to be expressed from a tetracycline-inducible promoter in di-cistronic configuration coexpressing GFP and the BVDV protein of interest. Thereafter, 293A cells were cotransfected with FseI-restricted transfer vector, and the ClaI-restricted Ad5/∆E1∆E3 viral DNA to generate recombinant viruses by in vivo homologous recombination between overlapping sequences of linearized transfer vectors pAdTR5-DC-GFPq, and Ad5/∆E1∆E3 viral DNA [25,26]. Adenoviral plaques were screened 10 to 15 days after cell transfection by monitoring basal GFP expression by fluorescence microscopy.…”

Section: Construction Of Advmentioning

confidence: 99%

“…Adenoviral plaques were screened 10 to 15 days after cell transfection by monitoring basal GFP expression by fluorescence microscopy. AdV were then purified by three further rounds of plaque isolation on BMAdE1 cells, and expanded as described [26]. Titers of the AdV stocks were estimated by a method based on the measurement of the GFP signal by cytofluorometry [25].…”

Section: Construction Of Advmentioning

confidence: 99%

“…The cells were replenished with the caspase inhibitors 24 h later, and collected at 48 h after infection with AdV for cytometry analysis. For HSV2-R1 and hsp70 inhibition assays, the cells were infected with both the BVDV AdV and an AdV expressing either HSV2-R1 (AdTR5-R1; AdV-HSV2-R1) [24] or hsp70 (AdTR5-hsp70-DC-BFP; AdV-hsp70) [26], and then tested at 48 h pi for apoptosis by cytometry analysis.…”

Section: Caspase-specific Peptide Inhibitors and The R1 Subunit Of Hmentioning

confidence: 99%

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The bovine viral diarrhea virus (BVDV) NS3 protein, when expressed alone in mammalian cells, induces apoptosis which correlates with caspase-8 and caspase-9 activation

2005

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-The bovine viral diarrhea virus (BVDV) strains exist as two biotypes, cytopathic (cp) and noncytopathic (ncp), according to their effects on tissue culture cells. It has been previously reported that cell death associated to cp BVDV in vitro is mediated by apoptosis. Here, experiments were conducted to determine the involvement of the NS3 protein in the induction of apoptosis. The NS3-and NS3∆50 (deleted from the NH2-terminal 50 amino acids)-cDNA encoding sequences of BVDV NADL cp reference strain were cloned into adenoviral vectors (AdV) from which the BVDV gene of interest could be expressed from a tetracycline-responsive promoter. A549tTA cells infected in vitro with NS3 or NS3∆50-expressing AdV showed cytopathic changes characterized by cell rounding and detachment, and nucleus chromatin condensation. DNA fragmentation assays, cytochrome c release, and activation of cellular caspases performed on these infected cells clearly correlated with the observed cytopathic changes with apoptosis. The BVDV NS3∆50-induced apoptotic process was inhibited by caspase-8-and -9-specific peptide inhibitors (Z-IETD-FMK and Z-LEHD-FMK). Furthermore, apoptosis was inhibited in cells expressing the R1 subunit of herpes simplex virus type 2 ribonucleotide reductase (HSV2-R1) or hsp70, two proteins which are known to inhibit apoptosis associated with caspase-8 activation and cytochrome c release-dependent caspase-9 activation, respectively. Given that HSV2-R1, a specific inhibitor of the caspase-8 activation pathway, efficiently suppressed apoptosis and also prevented caspase-9 activation, the overall results indicate that the BVDV NS3/NS3∆50 induces apoptosis initiated by caspase-8 activation and subsequent cytochrome c release-dependent caspase-9 activation. bovine viral diarrhea virus / NS3 protein / apoptosis / caspases

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Section: Cells and Virusesmentioning

confidence: 99%

Section: Construction Of Advmentioning

confidence: 99%

Section: Construction Of Advmentioning

confidence: 99%

Section: Caspase-specific Peptide Inhibitors and The R1 Subunit Of Hmentioning

confidence: 99%

See 2 more Smart Citations

The bovine viral diarrhea virus (BVDV) NS3 protein, when expressed alone in mammalian cells, induces apoptosis which correlates with caspase-8 and caspase-9 activation

2005

Self Cite

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“…To minimize the work involved in screening recombinant from parental viruses, reporter genes such as E. coli LacZ 15 or the green/blue fluorescent proteins (GFP/BFP) from A. victoria 16 can be used either in the viral genome as a negative screen, or in the transfer vector as a positive screen. Although useful, this approach still suffers from the intrinsic limitation that, in a library of several thousand clones, an even larger number of parental viruses would have to been screened out, a process which is fairly time-consuming.…”

Section: Cells Was Generated With 100% Of the Plaques Being Recombinamentioning

confidence: 99%

Adenovirus-based libraries: efficient generation of recombinant adenoviruses by positive selection with the adenovirus protease

Elahi¹,

Oualikene²,

Naghdi³

et al. 2002

Gene Ther

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A simple macroscopic model for the diffusion and adsorption kinetics of r‐adenovirus

Gilbert

Kamen

Bernier

et al. 2007

Biotech & Bioengineering

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The diffusion of viruses toward cells is a limiting step of the infection process. To be modeled correctly, this step must be evaluated in combination with the adsorption of the virus to the cell surface, which is a rapid but reversible step. In this paper, the recombinant adenovirus (rAd) diffusion and its adsorption to 293S cells in suspension were both measured and modeled. First, equilibrium experiments permitted to determine the number of receptors on the surface of 293S (R(T) = 3,500 cell(-1)) and the association constant (K(A) = 1.9 x 10(11) M(-1)) for rAd on these cells based on a simple monovalent adsorption model. Non-specific binding of the virus to the cell surface was not found to be significant. Second, total virus particle degradation rates between 5.2 x 10(-3) and 4.0 x 10(-2) min(-1) were measured at 37 degrees C in culture medium, but no significant virus degradation was observed at 4 degrees C. Third, free viral particle disappearance rates from a mixed suspension of virus and cells were measured at different virus concentrations. Experimental data were compared to a phenomenological dynamic model comprising both the diffusion and the adsorption steps. The diffusion to adsorption ratio, a fitted parameter, confirmed that the contact process of a virus with a cell is indeed diffusion controlled. However, the characteristic diffusion time constants obtained, based on a reversible adsorption model, were eightfolds smaller than those reported in the literature, based on diffusion models that assume irreversible adsorption.

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New adenovirus vectors for protein production and gene transfer

Cited by 27 publications

References 26 publications

The bovine viral diarrhea virus (BVDV) NS3 protein, when expressed alone in mammalian cells, induces apoptosis which correlates with caspase-8 and caspase-9 activation

The bovine viral diarrhea virus (BVDV) NS3 protein, when expressed alone in mammalian cells, induces apoptosis which correlates with caspase-8 and caspase-9 activation

Adenovirus-based libraries: efficient generation of recombinant adenoviruses by positive selection with the adenovirus protease

A simple macroscopic model for the diffusion and adsorption kinetics of r‐adenovirus

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