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Optimization of a gene electrotransfer procedure for efficient intradermal immunization with an hTERT-based DNA vaccine in mice.

Calvet CY, Thalmensi J, Liard C, Pliquet E, Bestetti T, Huet T, Langlade-Demoyen P, Mir LM - Mol Ther Methods Clin Dev (2014)

Bottom Line: Gene electrotransfer is the most efficient and safest non-viral gene transfer procedure and specific electrical parameters have been developed for several target tissues.In a second time, these parameters were tested for their potency to generate specific cellular CD8 immune responses against telomerase epitopes.These CD8 T-cells were fully functional as they secreted IFNγ and were endowed with specific cytotoxic activity towards target cells.

View Article: PubMed Central - PubMed

Affiliation: Univ Paris-Sud, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses , Villejuif, France ; CNRS, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses , Villejuif, France ; Gustave Roussy, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses , Villejuif, France.

ABSTRACT
DNA vaccination consists in administering an antigen-encoding plasmid in order to trigger a specific immune response. This specific vaccine strategy is of particular interest to fight against various infectious diseases and cancer. Gene electrotransfer is the most efficient and safest non-viral gene transfer procedure and specific electrical parameters have been developed for several target tissues. Here, a gene electrotransfer protocol into the skin has been optimized in mice for efficient intradermal immunization against the well-known telomerase tumor antigen. First, the luciferase reporter gene was used to evaluate gene electrotransfer efficiency into the skin as a function of the electrical parameters and electrodes, either non-invasive or invasive. In a second time, these parameters were tested for their potency to generate specific cellular CD8 immune responses against telomerase epitopes. These CD8 T-cells were fully functional as they secreted IFNγ and were endowed with specific cytotoxic activity towards target cells. This simple and optimized procedure for efficient gene electrotransfer into the skin using the telomerase antigen is to be used in cancer patients for the phase 1 clinical evaluation of a therapeutic cancer DNA vaccine called INVAC-1.

No MeSH data available.


Related in: MedlinePlus

Determination of the optimal HV pulse intensity for intradermally injected pCMV-luc electrotransfer in C57BL/6J mice using plate electrodes. Bioluminescence intensities were evaluated 2 days after pCMV-luc electrotransfer performed in C57BL/6J mice. One single HV pulse of different intensities in combination with one LV pulse of 140 V/cm were compared, n = 24 mice for pCMV-luc ID injection alone, n = 8 (from 4 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test with Dunn’s multiple comparison test.
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fig4: Determination of the optimal HV pulse intensity for intradermally injected pCMV-luc electrotransfer in C57BL/6J mice using plate electrodes. Bioluminescence intensities were evaluated 2 days after pCMV-luc electrotransfer performed in C57BL/6J mice. One single HV pulse of different intensities in combination with one LV pulse of 140 V/cm were compared, n = 24 mice for pCMV-luc ID injection alone, n = 8 (from 4 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test with Dunn’s multiple comparison test.

Mentions: Among the tested amplitudes, C57BL/6J mice electrotransferred with field amplitudes of 1,200 V/cm; 1,400 V/cm; or 1,600 V/cm presented the most significant enhancements of luciferase expression as compared to control mice (P < 0.001) (Figure 4). In particular, the highest median bioluminescence was obtained in the group treated with a field amplitude of 1,400 V/cm. There was also a better homogeneity in the results for this group as compared to other groups. However, there was no statistical difference between luciferase expressions obtained in the 1,200; 1,400; and 1,600 V/cm groups.


Optimization of a gene electrotransfer procedure for efficient intradermal immunization with an hTERT-based DNA vaccine in mice.

Calvet CY, Thalmensi J, Liard C, Pliquet E, Bestetti T, Huet T, Langlade-Demoyen P, Mir LM - Mol Ther Methods Clin Dev (2014)

Determination of the optimal HV pulse intensity for intradermally injected pCMV-luc electrotransfer in C57BL/6J mice using plate electrodes. Bioluminescence intensities were evaluated 2 days after pCMV-luc electrotransfer performed in C57BL/6J mice. One single HV pulse of different intensities in combination with one LV pulse of 140 V/cm were compared, n = 24 mice for pCMV-luc ID injection alone, n = 8 (from 4 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test with Dunn’s multiple comparison test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362362&req=5

fig4: Determination of the optimal HV pulse intensity for intradermally injected pCMV-luc electrotransfer in C57BL/6J mice using plate electrodes. Bioluminescence intensities were evaluated 2 days after pCMV-luc electrotransfer performed in C57BL/6J mice. One single HV pulse of different intensities in combination with one LV pulse of 140 V/cm were compared, n = 24 mice for pCMV-luc ID injection alone, n = 8 (from 4 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test with Dunn’s multiple comparison test.
Mentions: Among the tested amplitudes, C57BL/6J mice electrotransferred with field amplitudes of 1,200 V/cm; 1,400 V/cm; or 1,600 V/cm presented the most significant enhancements of luciferase expression as compared to control mice (P < 0.001) (Figure 4). In particular, the highest median bioluminescence was obtained in the group treated with a field amplitude of 1,400 V/cm. There was also a better homogeneity in the results for this group as compared to other groups. However, there was no statistical difference between luciferase expressions obtained in the 1,200; 1,400; and 1,600 V/cm groups.

Bottom Line: Gene electrotransfer is the most efficient and safest non-viral gene transfer procedure and specific electrical parameters have been developed for several target tissues.In a second time, these parameters were tested for their potency to generate specific cellular CD8 immune responses against telomerase epitopes.These CD8 T-cells were fully functional as they secreted IFNγ and were endowed with specific cytotoxic activity towards target cells.

View Article: PubMed Central - PubMed

Affiliation: Univ Paris-Sud, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses , Villejuif, France ; CNRS, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses , Villejuif, France ; Gustave Roussy, Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses , Villejuif, France.

ABSTRACT
DNA vaccination consists in administering an antigen-encoding plasmid in order to trigger a specific immune response. This specific vaccine strategy is of particular interest to fight against various infectious diseases and cancer. Gene electrotransfer is the most efficient and safest non-viral gene transfer procedure and specific electrical parameters have been developed for several target tissues. Here, a gene electrotransfer protocol into the skin has been optimized in mice for efficient intradermal immunization against the well-known telomerase tumor antigen. First, the luciferase reporter gene was used to evaluate gene electrotransfer efficiency into the skin as a function of the electrical parameters and electrodes, either non-invasive or invasive. In a second time, these parameters were tested for their potency to generate specific cellular CD8 immune responses against telomerase epitopes. These CD8 T-cells were fully functional as they secreted IFNγ and were endowed with specific cytotoxic activity towards target cells. This simple and optimized procedure for efficient gene electrotransfer into the skin using the telomerase antigen is to be used in cancer patients for the phase 1 clinical evaluation of a therapeutic cancer DNA vaccine called INVAC-1.

No MeSH data available.


Related in: MedlinePlus