Comparison of bicistronic retroviral vectors containing internal ribosome entry sites (IRES) using expression of human interleukin-12 (IL-12) as a readout

Harries, Mark; Phillipps, Neil; Prentice, G; Collins, Mary

doi:10.1002/1521-2254(200007/08)2:4<243::aid-jgm115>3.0.co;2-q

Cited by 19 publications

(4 citation statements)

References 60 publications

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“…36,21 Our data confirm that this approach leads to production of high levels of biologically functional IL12 with little evident impairment of T-cell behavior.…”

Section: Discussionsupporting

confidence: 74%

A strategy for treatment of Epstein–Barr virus-positive Hodgkin's disease by targeting interleukin 12 to the tumor environment using tumor antigen-specific T cells

et al. 2003

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Adoptive immunotherapy with Epstein-Barr virus (EBV)-specific cytotoxic T cells (CTL) is effective for the prophylaxis and treatment of EBV-induced lymphoma in hematopoietic stem cell recipients. However, in EBV-positive Hodgkin's disease (HD) the efficacy of adoptively transferred EBV-specific CTL may be limited by tumor-derived immunosuppressive factors, such as T-cell growth factor (TGF) b, interleukin (IL)13 and the chemokine TARC. Local delivery of IL12 to tumor sites by tumor-specific CTL could provide direct antitumor effects and overcome the CTL-inhibitory effects of the Th2 tumor environment while avoiding the systemic toxicity of recombinant IL12. EBV-specific CTL transduced with a retrovirus vector expressing the p40 and p35 subunits of IL12 as a single molecule (Flexi-IL12), produced IL12 following antigenic stimulation. This resulted in an elevated production of Th1 cytokines, including interferon g and tumor necrosis factor a, and a reduction in the Th2 cytokines IL4 and IL5. Flexi-IL12-transduced CTL resisted the antiproliferative and anticytotoxic effects of exogenous TGFb, likely by antagonizing the TGFb-induced downregulation of the Th1 transcriptional factor T-bet. In addition, Flexi-IL12-transduced CTL demonstrated a proliferative advantage in the presence of inhibitory supernatants from HD-derived cell lines. Tumor-specific, Flexi-IL12-transduced EBV-specific CTL should have a functional advantage over unmodified CTL, particularly in the presence of the adverse Th2 cytokine environment produced by Hodgkin tumor cells.

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“…36,21 Our data confirm that this approach leads to production of high levels of biologically functional IL12 with little evident impairment of T-cell behavior.…”

Section: Discussionsupporting

confidence: 74%

A strategy for treatment of Epstein–Barr virus-positive Hodgkin's disease by targeting interleukin 12 to the tumor environment using tumor antigen-specific T cells

et al. 2003

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“…To address this issue, several groups have sought to identify alternative cellular and viral IRES elements that initiate protein expression more efficiently than EMCV IRES. 15,[19][20][21][22][23][24][25][26] Cellular IRES elements such as eukaryotic initiation factor 4G, 15 immunoglobulin heavy chain binding protein, 15 c-myc proto-oncogene, 15 vascular endothelial growth factor 15 and fibroblast growth factor-1 27 IRES have been reported to show higher efficiency than EMCV IRES. In this study, we searched to identify an IRES(s) with higher translation efficiency than the EMCV IRES both in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

In vitro and in vivo comparison of viral and cellular internal ribosome entry sites for bicistronic vector expression

et al. 2011

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Bicistronic vectors are essential to achieve efficient expression of multiple genes in gene therapy protocols and biomedical applications. Internal ribosome entry site (IRES) elements have been utilized to initiate expression of an additional protein from a bicistronic vector. The IRES element commonly used in current bicistronic vectors originates from the encephalomyocarditis virus (EMCV). As IRES-mediated translation is dependent on availability of IRES trans-acting factors, which vary between cell types and species, adequate gene expression from the EMCV IRES element is not always achieved. To identify a novel IRES element that mediates gene expression consistently with a higher efficiency than the EMCV IRES, we tested 13 bicistronic reporter constructs containing different viral and cellular IRES elements. The in vitro screening in human and mouse fibroblast and hepatocarcinoma cells revealed that the vascular endothelial growth factor and type 1 collagen-inducible protein (VCIP) IRES was the only IRES element that directed translation more efficiently than the EMCV IRES in all cell lines. Furthermore, the VCIP IRES initiated greater reporter expression levels than the EMCV IRES in transfected mouse livers. These results suggest that VCIP-IRES containing vectors improve gene expression compared with those harboring an EMCV-IRES. This could increase the potential benefits of bicistronic vectors for experimental and therapeutic purposes.

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“…[24][25][26] However, the efficiency of IRES-dependent translation may vary in different cells and tissues [27][28][29] and IRES-dependent second gene expression can be significantly lower than cap-dependent first gene expression in bicistronic vectors. 30,31 Furthermore, in consideration of the low insert capacity of rAAV vectors, an element promoting bicistronic expression but being shorter than the IRES may be preferred.…”

Section: Introductionmentioning

confidence: 99%

Recombinant AAV vectors containing the foot and mouth disease virus 2A sequence confer efficient bicistronic gene expression in cultured cells and rat substantia nigra neurons

et al. 2001

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Recombinant adeno-associated viruses (rAAVs) are promising vectors for gene therapy since they efficiently and stably transduce a variety of tissues of immunocompetent animals. The major disadvantage of rAAVs is their limited capacity to package foreign DNA (р5 kb). Often, co-expression of two or more genes from a single viral vector is desirable to achieve maximal therapeutic efficacy or to track transduced cells in vivo by suitable reporter genes. The internal ribosome entry site (IRES) sequence of encephalomyocarditis virus has been widely used to construct bicistronic viral vectors. However, the IRES is rather long and IRES-mediated translation can be relatively inefficient when compared with capdependent translation. As an alternative to the IRES for in vivo gene expression, we studied the 16 amino-acid long 2A peptide of foot and mouth disease virus (FMDV). The 2A peptide mediates the primary cis-'cleavage' of the FMDV polyprotein in a cascade of processing events that ultimately

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Comparison of bicistronic retroviral vectors containing internal ribosome entry sites (IRES) using expression of human interleukin-12 (IL-12) as a readout

Cited by 19 publications

References 60 publications

A strategy for treatment of Epstein–Barr virus-positive Hodgkin's disease by targeting interleukin 12 to the tumor environment using tumor antigen-specific T cells

A strategy for treatment of Epstein–Barr virus-positive Hodgkin's disease by targeting interleukin 12 to the tumor environment using tumor antigen-specific T cells

In vitro and in vivo comparison of viral and cellular internal ribosome entry sites for bicistronic vector expression

Recombinant AAV vectors containing the foot and mouth disease virus 2A sequence confer efficient bicistronic gene expression in cultured cells and rat substantia nigra neurons

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