Highly Efficient Lentiviral Gene Transfer in CD34
 +
 and CD34
 +
 /38
 −
 /lin
 −
 Cells from Mobilized Peripheral Blood after Cytokine Prestimulation

Geronimi, Fabien

doi:10.1634/stemcells.21-4-472

Cited by 21 publications

(13 citation statements)

References 51 publications

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“…The use of a low MOI is warranted for clinical trials, as the risk of insertional mutagenesis, also for lentiviral vectors, is present. A number of studies show that a higher MOI is needed for MPB than for cord blood to reach a sufficient transduction efficiency and that with an MOI of 100, gene transfer rates were obtained with a reasonable number of integration sites (a figure in the neighborhood of three to four copies per cell), thus limiting the risk of insertional oncogenesis [19]. In the present protocol, with an MOI of 50 we obtained a reasonable proportion of transduced cells, in which only 0.4 vector copies are integrated in the genome.…”

Section: Discussionmentioning

confidence: 99%

Sustained Long-Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Third-Generation Lentiviral Vectors

Tesio¹,

Gammaitoni²,

Gunetti³

et al. 2008

Stem Cells

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Section: Discussionmentioning

confidence: 99%

Sustained Long-Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Third-Generation Lentiviral Vectors

Tesio¹,

Gammaitoni²,

Gunetti³

et al. 2008

Stem Cells

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“…35 Previously, we have successfully used an introndeleted version of this human promoter in human CD34 þ cells. 36,37 However, this short version was too weak to allow an efficient in vivo chemoprotection of transduced HSCs (preliminary data not shown). This study confirms the interest of the intron to reach a level of gene expression obtained as high as with a strong viral ESp but without acting as long-distance enhancers.…”

Section: Discussionmentioning

confidence: 99%

Murine Retroviral but not Human Cellular Promoters Induce In Vivo Erythroid-specific Deregulation that can be Partially Prevented by Insulators

et al. 2007

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We are developing lentiviral vectors for gene therapy of red blood cell disorders that co-express a transgene in an erythroid-specific manner and the O(6)-methylguanine-DNA-methyltransferase (MGMT) selective gene in a constitutive way. We report that transduction of murine hematopoietic stem cells (HSCs) with a human phosphoglycerate kinase promoter-based vector at low multiplicity of infection (MOI) does not result in a selective in vivo expansion in the presence of alkylating agents. In contrast, by replacing this cellular promoter with the powerful retroviral-derived myeloproliferative sarcoma virus enhancer, negative control region-deleted, dl587rev primer-binding site substituted promoter, the vector allowed efficient chemoprotection of transduced HSCs at low MOI. However, this promoter interacted with the erythroid HS40/ankyrin enhancer/promoter driving green fluorescent protein, leading to an unexpected loss of erythroid specificity. A partial restoration of tissue-specific expression was obtained by interposition of insulator sequences between the expression units. Alternatively, we found that the strong human cellular elongation factor1-alpha promoter allows similar chemoprotection but without any deregulation of the erythroid-specific promoter in the absence of insulators. These data demonstrate that the level of in vivo deregulation induced by a promoter is not correlated with its transcriptional activity.

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“…Expression of EGFP, CAT, and CuZn-SOD was driven by the human phosphoglycerate kinase promoter. Lentiviral particles were produced by transient transfection of 293T cells as described previously (27). The following plasmids were used: a packaging plasmid (pCMV⌬8.91), a vesicular somatitis virus glycoprotein (VSV-G) envelope plasmid pMD.G, and a transducing vector (TPW, TPEW, TPCATW, TPCuZnSODW).…”

Section: Methodsmentioning

confidence: 99%

Protective Effects of Catalase Overexpression on UVB-induced Apoptosis in Normal Human Keratinocytes

Rezvani

Mazurier

Cario‐André

et al. 2006

Journal of Biological Chemistry

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107

112

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UV-induced apoptosis in keratinocytes is a highly complex process in which various molecular pathways are involved. These include the extrinsic pathway via triggering of death receptors and the intrinsic pathway via DNA damage and reactive oxygen species (ROS) formation. In this study we investigated the effect of catalase and CuZn-superoxide dismutase (SOD) overexpression on apoptosis induced by UVB exposure at room temperature or 4°C on normal human keratinocytes. Irradiation at low temperature reduced UV-induced apoptosis by 40% in normal keratinocytes independently of any change in p53 and with a decrease in caspase-8 activation. Catalase overexpression decreased apoptosis by 40% with a reduction of caspase-9 activation accompanied by a decrease in p53. Keeping cells at low temperature and catalase overexpression had additive effects. CuZn-SOD overexpression had no significant effect on UVB-induced apoptosis. UVB induced an increase in ROS levels at two distinct stages: immediately following irradiation and around 3 h after irradiation. Catalase overexpression inhibited only the late increase in ROS levels. We conclude that catalase overexpression has a protective role against UVB irradiation by preventing DNA damage mediated by the late ROS increase.

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Highly Efficient Lentiviral Gene Transfer in CD34 ⁺ and CD34 ⁺ /38 ⁻ /lin ⁻ Cells from Mobilized Peripheral Blood after Cytokine Prestimulation

Abstract: ABSTRACT

Cited by 21 publications

References 51 publications

Sustained Long-Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Third-Generation Lentiviral Vectors

Sustained Long-Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Third-Generation Lentiviral Vectors

Murine Retroviral but not Human Cellular Promoters Induce In Vivo Erythroid-specific Deregulation that can be Partially Prevented by Insulators

Protective Effects of Catalase Overexpression on UVB-induced Apoptosis in Normal Human Keratinocytes

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Highly Efficient Lentiviral Gene Transfer in CD34 + and CD34 + /38 − /lin − Cells from Mobilized Peripheral Blood after Cytokine Prestimulation

Abstract: ABSTRACT

Cited by 21 publications

References 51 publications

Sustained Long-Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Third-Generation Lentiviral Vectors

Sustained Long-Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Third-Generation Lentiviral Vectors

Murine Retroviral but not Human Cellular Promoters Induce In Vivo Erythroid-specific Deregulation that can be Partially Prevented by Insulators

Protective Effects of Catalase Overexpression on UVB-induced Apoptosis in Normal Human Keratinocytes

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Highly Efficient Lentiviral Gene Transfer in CD34 ⁺ and CD34 ⁺ /38 ⁻ /lin ⁻ Cells from Mobilized Peripheral Blood after Cytokine Prestimulation