DNA electrotransfer after intradermal injection for immunisation
(2008)
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- Abstract
- The specific immunologic environment of the skin provides both innate and adaptive immunities and efficiently protects the individual from external aggressions; making the skin an attractive organ for the delivery of DNA vaccines. However, the development of such vaccines requires appropriate delivery technologies. Electrotransfer is one of the most efficient methods of in vivo non-viral gene transfer. The introduction reviews the literature and gives information about the main fields evoked in the thesis: the skin, methods for DNA transfer in the skin, electrotransfer and DNA vaccines. The aim of this work was to optimise immunisation by intradermal DNA electrotransfer. First, we optimised the delivery method itself. The effects of the injection method and the DNA dose on gene expression after intradermal DNA electrotransfer were studied. Pre-treatments were evaluated to enhance DNA diffusion in the skin. Then, DNA immunisation with a weakly immunogenic model antigen, luciferase, was investigated. The immune response after intradermal DNA electrotransfer was compared with immunisation to the luciferase recombinant protein. Finally, the influence of the delivery route was studied. Second, we tried to find adjuvants to modulate and/or enhance the immune response after intradermal DNA electrotransfer. Four potential adjuvants, acting on different targets were studied: hyaluronidase, imiquimod, monophosphoryl lipid A and tape stripping. Finally, we worked on the optimisation of the plasmid vector. We investigated if it was possible to induce immune response when only some specific cells of the skin were targeted. Specific promoters of cutaneous cells were cloned and we investigated if fascin, keratin14 or CD11c promoter plasmids were able to induce immune response. We also followed kinetic of luciferase expression after DNA electrotransfer in the muscle and in the skin of a small eukaryotic expression vector which was devoid of antibiotic resistance markers and optimised for gene therapy.
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Vandermeulen, G. (2008). DNA electrotransfer after intradermal injection for immunisation. https://hdl.handle.net/2078.5/112049