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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

Choice of the best HV-LV pulses combination for pCMV-luc dermal electrotransfer and INVAC-1–mediated ID vaccination. (a) Bioluminescence obtained in C57BL/6J mice 2 days after pCMV-luc ID injection upon various HV-LV pulses combinations, n = 30 mice for pCMV-luc ID injection alone and n = 6 (from 3 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP delivered onto the skin. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with INVAC-1 according to various combinations of HV-LV pulses, n = 8 mice for PBS immunization control and n = 5 mice for INVAC-1–mediated immunization. (c) Comparison of the bioluminescence intensities in C57BL/6J mice in which EGT was performed 2 days before with P9 or Pd, n = 4 mice for pCMV-luc ID injection alone, n = 8 (from four mice, two treatments per mouse) for pCMV-luc ID injection+EP. (d) Comparison of the frequency of hTERT-specific IFNγ+ CD8 T-cells in C57BL/6J mice immunized intradermally 14 days before by using INVAC-1 combined with P9 or Pd, n = 4 mice for PBS immunization control and n = 6 mice for INVAC-1–mediated immunization. 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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fig5: Choice of the best HV-LV pulses combination for pCMV-luc dermal electrotransfer and INVAC-1–mediated ID vaccination. (a) Bioluminescence obtained in C57BL/6J mice 2 days after pCMV-luc ID injection upon various HV-LV pulses combinations, n = 30 mice for pCMV-luc ID injection alone and n = 6 (from 3 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP delivered onto the skin. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with INVAC-1 according to various combinations of HV-LV pulses, n = 8 mice for PBS immunization control and n = 5 mice for INVAC-1–mediated immunization. (c) Comparison of the bioluminescence intensities in C57BL/6J mice in which EGT was performed 2 days before with P9 or Pd, n = 4 mice for pCMV-luc ID injection alone, n = 8 (from four mice, two treatments per mouse) for pCMV-luc ID injection+EP. (d) Comparison of the frequency of hTERT-specific IFNγ+ CD8 T-cells in C57BL/6J mice immunized intradermally 14 days before by using INVAC-1 combined with P9 or Pd, n = 4 mice for PBS immunization control and n = 6 mice for INVAC-1–mediated immunization. Bars represent median values. *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test with Dunn’s multiple comparison test.

Mentions: The P4, P8, and P9 combinations of HV-LV pulses generated the highest median bioluminescence intensities (Figure 5a). All of these three combinations displayed very high statistical differences when compared to pCMV-luc ID injection without EP (P < 0.001). In particular, P9 showed the best median bioluminescence intensity, the highest value for the minimum bioluminescence intensity and the lowest point dispersion.


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)

Choice of the best HV-LV pulses combination for pCMV-luc dermal electrotransfer and INVAC-1–mediated ID vaccination. (a) Bioluminescence obtained in C57BL/6J mice 2 days after pCMV-luc ID injection upon various HV-LV pulses combinations, n = 30 mice for pCMV-luc ID injection alone and n = 6 (from 3 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP delivered onto the skin. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with INVAC-1 according to various combinations of HV-LV pulses, n = 8 mice for PBS immunization control and n = 5 mice for INVAC-1–mediated immunization. (c) Comparison of the bioluminescence intensities in C57BL/6J mice in which EGT was performed 2 days before with P9 or Pd, n = 4 mice for pCMV-luc ID injection alone, n = 8 (from four mice, two treatments per mouse) for pCMV-luc ID injection+EP. (d) Comparison of the frequency of hTERT-specific IFNγ+ CD8 T-cells in C57BL/6J mice immunized intradermally 14 days before by using INVAC-1 combined with P9 or Pd, n = 4 mice for PBS immunization control and n = 6 mice for INVAC-1–mediated immunization. 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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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4362362&req=5

fig5: Choice of the best HV-LV pulses combination for pCMV-luc dermal electrotransfer and INVAC-1–mediated ID vaccination. (a) Bioluminescence obtained in C57BL/6J mice 2 days after pCMV-luc ID injection upon various HV-LV pulses combinations, n = 30 mice for pCMV-luc ID injection alone and n = 6 (from 3 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP delivered onto the skin. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with INVAC-1 according to various combinations of HV-LV pulses, n = 8 mice for PBS immunization control and n = 5 mice for INVAC-1–mediated immunization. (c) Comparison of the bioluminescence intensities in C57BL/6J mice in which EGT was performed 2 days before with P9 or Pd, n = 4 mice for pCMV-luc ID injection alone, n = 8 (from four mice, two treatments per mouse) for pCMV-luc ID injection+EP. (d) Comparison of the frequency of hTERT-specific IFNγ+ CD8 T-cells in C57BL/6J mice immunized intradermally 14 days before by using INVAC-1 combined with P9 or Pd, n = 4 mice for PBS immunization control and n = 6 mice for INVAC-1–mediated immunization. Bars represent median values. *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test with Dunn’s multiple comparison test.
Mentions: The P4, P8, and P9 combinations of HV-LV pulses generated the highest median bioluminescence intensities (Figure 5a). All of these three combinations displayed very high statistical differences when compared to pCMV-luc ID injection without EP (P < 0.001). In particular, P9 showed the best median bioluminescence intensity, the highest value for the minimum bioluminescence intensity and the lowest point dispersion.

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