Polyethylenimine-mediated transfection of human monocytes with the IFN-γ gene: an approach for cancer adoptive immunotherapy

Ringenbach, Laurence; Bohbot, Alain; Tiberghien, Pierre; Oberling, F; Feugeas, Olivier

doi:10.1038/sj.gt.3300756

Cited by 21 publications

(18 citation statements)

References 28 publications

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“…1 A variety of cell types such as monocytes, 2 dendritic cells, 3 myoblast cells, 4 and hepatocytes 5 were studied as target cells for PEI-mediated gene transfection. In vivo, PEI was shown to be an efficient transfection vector in several organs such as the kidney, liver, and lung.…”

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Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes

Kim

Yoon

et al. 2001

Gene Ther

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Polyethylenimine (PEI) Recently, polyethylenimine (PEI) has drawn attention as a versatile, inexpensive, and useful nonviral transfection vector. 1 A variety of cell types such as monocytes, 2 dendritic cells, 3 myoblast cells, 4 and hepatocytes 5 were studied as target cells for PEI-mediated gene transfection. In vivo, PEI was shown to be an efficient transfection vector in several organs such as the kidney, liver, and lung. [6][7][8] However, these studies are mostly focused on the expression of the PEI/DNA limited to target organs in vivo or specific cell types in vitro. Although the knowledge of biodistribution fates of plasmid DNA may reveal the feasibility of the cellular level targeting in vivo, there is little understanding of the overall and quantitative organ distribution of plasmid DNA after administration in PEI complexes. Moreover, the safety of PEI/DNA complexes after repeated dosing has not been well documented, despite the potential requirement for repeated administration due to the episomal property of plasmids.In this study, we tested the distribution fate and safety of plasmid DNA after administering DNA in PEI complexes. Quantitative and competitive polymerase chain reaction (PCR) was employed for the assay of plasmid DNA in the biological samples. Here, we report that in comparison to naked DNA, the organ distribution was higher and remarkably prolonged after administration of plasmid DNA in PEI complexes, and that once a week dosing of PEI/DNA complexes over 3 weeks did not alter the histology of major organs such as the liver, lung, and kidney.PEI/DNA complexes with PEI nitrogen to DNA phosphate ratio (N/P ratio) of 10:1 were prepared in 5% glucose using 25 kDa PEI (Aldrich, St Quentin, France). The N/P ratio of 10:1 was chosen since it was shown to efficiently transfect cells in vivo. 9 The levels of plasmid DNA in biological samples were determined using competitive and quantitative PCR. Quantitative PCR is based on co-amplification of the sample template together with various amounts of internal standard (IS) competitor sharing with the target the primer recognition sites, but differing in size. The competitor was constructed to share the same sense and antisense primer used for target amplification. 10 As an example, Figure 1 shows the gel bands and internal standard curves obtained after running competitive PCR of DNA extracts from the liver 6 h after injection of naked DNA or PEI/DNA complexes. The IS for quantitative PCR was the 188-bp deleted mutant of pCMV␤. The construction of the competitor and PCR conditions were described previously. 10 The PCR products were 449 bp and 261 bp for the target and the IS, respectively. The band densities of the target gene PCR product diminished as the amounts of IS increased (Figure 1a and b). The quantitation of gel band density by densitometry generated internal standard curves of high linearity (Figure 1c). The curves also show that we could detect as low as fg-level plasmids by quantitative PCR. The amounts of plasmid DNA in the samples we...

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confidence: 99%

Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes

Kim

Yoon

et al. 2001

Gene Ther

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“…It is known MDP can be considered a functional lipopoly-saccharides analogue (LPS) (Hoffmann et al 1999). Among other actions, MDP stimulates the phagocytic activity of macrophages, microglia and Schwann cells (Parant et al 1992, Ringerbach et al 1998. Local MDP injection into the site of spinal cord injury was shown to improve behavioral outcome.…”

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confidence: 99%

Pharmacological immunomodulation enhances peripheral nerve regeneration

et al. 2007

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To assess the effect of N-Acetylmuramyl-L-Alanyl-D-Isoglutamine MDP topically administrated on the regenerating peripheral neurons, twelve male C57BL/6J adult mice were equally distributed into three groups. Four mice underwent unilateral sciatic nerve transection and polyethylene tubulization, with a 4mm gap between the proximal and distal nerve stumps and were implanted with collagen + PBS (COL). Other four animals underwent the same surgical procedure but received collagen + MDP (COL/MDP) inside the prosthesis. Four animals were not operated and served as control group (NOR). After 4 weeks, the regenerated nerve cables were processed for total myelinated axon counting and myelinated fiber diameter measurement. The L5 dorsal root ganglion (DRG) was also removed and sectioned for sensory neurons counting and measurement. The results revealed significant difference (p<0.05) in axonal counting among the groups NOR (4,355±32), COL (1,869±289) and COL/MDP (2,430±223). There was a significant reduction in the axonal diameter in the operated groups (COL=3.38µm±1.16 and COL/MDP=3.54µm±1.16) compared to NOR (6.19µm±2.45). No difference was found in the number of DRG neurons between the experimental groups (COL=564±51; COL/MDP=514±56), which presented fewer sensory neurons compared to NOR (1,097±142). Data obtained indicate that locally applied MDP stimulates peripheral nerve regeneration in mice.

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“…Activated Mf are important to host defence because they express antimicrobial, antitumor and immunostimulatory activities, and IFNg is a well-known Mf activator. Studies in which DCs, 2 human monocytes, 6 and murine Mf 7 were engineered to express the IFNg gene demonstrated the relevance of this cytokine in cellular gene therapy protocols for treatment of cancer and infection.…”

Section: Activation Of Macrophage Vectors S Pastorino Et Almentioning

confidence: 99%

“…Translational application requires the manipulation of primary cells to be used as a source of Mf. 33 The possibilities of transducing bone marrow stem cells and to differentiate them into Mf or of engineering human mononuclear cells in vitro are reasonable approaches to obtain engineered Mf, 5,6,34 and experiments are underway to address these issues.…”

Section: Activation Of Macrophage Vectors S Pastorino Et Almentioning

confidence: 99%

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Picolinic acid- or desferrioxamine-inducible autocrine activation of macrophages engineered to produce IFNγ: an approach for gene therapy

et al. 2004

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Macrophage (Mf)-based vectors are highly mobile cellular shuttles designed to deliver therapeutic genes within the tissues. We engineered a mouse Mf cell line to express the murine interferon-g (IFNg) under the control of an inducible promoter containing the hypoxia-responsive element, which can be triggered by hypoxia and other stimuli. We show that this Mf vector can be induced to produce IFNg under normoxic conditions by stimulation with picolinic acid (PA), a catabolite of tryptophan, or desferrioxamine (DFX), an ironchelating drug. The Mf vector responds to IFNg with the induction of IRF-1 and of other IFNg-inducible genes, the expression of Ia antigens and induction of phagocytic activity.Inducible nitric oxygen synthase gene expression, nitric oxide production, as well as TNFa secretion were enhanced by PA or DFX as the sole stimuli. None of the above responses could be triggered individually by PA or DFX in control, normal Mf, indicating that the Mf vector overcame the need for costimulatory molecules derived from the immune system for its full activation. Furthermore, we demonstrate that extracellular iron can downregulate such response, thereby identifying an additional tool for the fine tuning of the Mf vector response to stimulation.

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Polyethylenimine-mediated transfection of human monocytes with the IFN-γ gene: an approach for cancer adoptive immunotherapy

Cited by 21 publications

References 28 publications

Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes

Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes

Pharmacological immunomodulation enhances peripheral nerve regeneration

Picolinic acid- or desferrioxamine-inducible autocrine activation of macrophages engineered to produce IFNγ: an approach for gene therapy

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