Gene transfer in humans using a conditionally replicating lentiviral vector

Levine, Bruce L.; Humeau, Laurent; Boyer, Jean; MacGregor, Rob-Roy; Rebello, Tessio; Lü, Xiaobin; Binder, Gwendolyn K.; Slepushkin, Vladimir; Lemiale, Franck; Mascola, John R.; Bushman, Frederic D.; Dropulić, Boro; June, Carl H.

doi:10.1073/pnas.0608138103

Cited by 445 publications

(372 citation statements)

References 21 publications

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“…The fact that TCR expression in PBL was generally less than one quarter of the activity in T-cell lines lead to the continued refinement in vector design such that all current TCR vectors now produce robust expression in transduced primary human lymphocytes. Recently, Levine et al 53 reported on a phase 1 clinical trial using lentiviral vectors that demonstrated efficient and safe gene delivery to patients' T cells with good persistence in vivo. The novel bicistronic lentiviral vectors harboring specific antitumor TCR described herein provide the opportunity not only for sustained transgene expression but also new methodologies to modify T lymphocytes at a less differentiated stage, and may have important applications in cancer immunotherapy.…”

Section: Discussionmentioning

confidence: 99%

Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition

et al. 2008

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In human gene therapy applications, lentiviral vectors may have advantages over g-retroviral vectors in several areas, including the ability to transduce nondividing cells, resistance to gene silencing and a potentially safer integration site profile. However, unlike g-retroviral vectors it has been problematic to drive the expression of multiple genes efficiently and coordinately with approaches such as internal ribosome entry sites or dual promoters. Using different 2A peptides, lentiviral vectors expressing two-gene T-cell receptors directed against the melanoma differentiation antigens gp100 and MART-1 were constructed. We demonstrated that addition of amino-acid spacer sequences (GSG or SGSG) before the 2A sequence is a prerequisite for efficient synthesis of biologically active T-cell receptors and that addition of a furin cleavage site followed by a V5 peptide tag yielded optimal T-cell receptor gene expression. Furthermore, we determined that the furin cleavage site was recognized in lymphocytes and accounted for removal of residual 2A peptides at the post-translational level with an efficiency of 20-30%, which could not be increased by addition of multiple furin cleavage sites. The novel bicistronic lentiviral vector developed herein afforded robust antimelanoma activities to engineered peripheral blood lymphocytes, including cytokine secretion, cell proliferation and lytic activity. Such optimal vectors may have immediate applications in cancer gene therapy.

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Section: Discussionmentioning

confidence: 99%

Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition

et al. 2008

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“…[1][2][3][4] A number of applications, such as the cure of polygenetic diseases as well as adoptive T-cell transfer and generation of induced pluripotent stem cells require more complex vector architectures capable of coexpressing two or more transgenes from the same vector backbone. 5,6 The most commonly used coexpression strategies are based on 2A cleavage or internal ribosome entry sites (IRES).…”

Section: Introductionmentioning

confidence: 99%

Mechanisms controlling titer and expression of bidirectional lentiviral and gammaretroviral vectors

et al. 2009

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Bidirectional lentiviral vectors mediate expression of two or more cDNAs from a single internal promoter. In this study, we examined mechanisms that control titer and expression properties of this vector system. To address whether the bidirectional design depends on lentiviral (LV) backbone components, especially the Rev/Rev responsive element (RRE) system, we constructed similar expression cassettes for LV and gammaretroviral (GV) vectors. Bidirectional expression levels could be adjusted by the use of different internal promoters. Furthermore, removal of the constitutive RNA transport element of Mason-Pfizer monkey virus, used in first generation bidirectional LV vectors, improved gene expression. Titers of bidirectional vectors were B10-fold reduced in comparison to unidirectional vectors, independent of the Rev/RRE interaction. We reasoned that titer reductions were due to the formation of interfering double-stranded RNA in packaging cells. Indeed, cotransfection of Nodamuravirus B2 protein, an RNA interference suppressor, increased bidirectional vector titers at least fivefold. We validated the potential of high titer bidirectional vectors by coexpressing a fluorescent marker with O 6 -methylguanine-DNA methyltransferase from integrating, or with Cre recombinase from integrating and non-integrating GV and LV backbones. This allowed for the tracking of chemoprotected and recombined cells by fluorescence marker expression.

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“…47 Early replication-competent lentivirus could be generated in vitro by recombination of vector plasmids or in vivo by mobilization of vector DNA in the presence of other infectious lentivirus such as HIV. 44 Currently used self-inactivating lentiviral vectors generated from three-or four-plasmid transfections are substantially less susceptible to generation of recombinant virus; furthermore, transgene expression can be driven from a heterologous internal promoter, preventing in vivo recombination 48 and enhancing long-term stable expression. To date, there have been no cases of replication-competent virus identified in cell products or patients.…”

Section: Replication Competent Virusmentioning

confidence: 99%

The Flipside of the Power of Engineered T Cells: Observed and Potential Toxicities of Genetically Modified T Cells as Therapy

2017

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Gene transfer in humans using a conditionally replicating lentiviral vector

Cited by 445 publications

References 21 publications

Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition

Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition

Mechanisms controlling titer and expression of bidirectional lentiviral and gammaretroviral vectors

The Flipside of the Power of Engineered T Cells: Observed and Potential Toxicities of Genetically Modified T Cells as Therapy

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