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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

Comparison of pCMV-luc gene transfer into the dermis and INVAC-1-mediated ID vaccination efficiencies with or without EP using plate electrodes. (a) Representation of bioluminescence intensities in C57BL/6J mice 2 days after pCMV-luc ID injection followed or not by EP, n = 5 mice for pCMV-luc ID injection alone, n = 10 (from 5 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with 25 µg of INVAC-1 followed or not by EP, n = 8 both for INVAC-1 ID injection alone or n = 6 for INVAC-1 ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, Mann–Whitney–Wilcoxon test.
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fig1: Comparison of pCMV-luc gene transfer into the dermis and INVAC-1-mediated ID vaccination efficiencies with or without EP using plate electrodes. (a) Representation of bioluminescence intensities in C57BL/6J mice 2 days after pCMV-luc ID injection followed or not by EP, n = 5 mice for pCMV-luc ID injection alone, n = 10 (from 5 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with 25 µg of INVAC-1 followed or not by EP, n = 8 both for INVAC-1 ID injection alone or n = 6 for INVAC-1 ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, Mann–Whitney–Wilcoxon test.

Mentions: C57BL/6J mice were injected intradermally on both flanks with either pCMV-luc (encoding luciferase) or INVAC-1 (encoding a modified form of hTERT) plasmids, followed or not by the application of EP (1 HV pulse of 1,000 V/cm and of 100 µs followed 1,000 ms later by 1 LV pulse of 140 V/cm and of 400 ms).1 Erythema was observed neither after mice shaving nor during or after the EGT procedure. Two parameters were measured after EGT according to the plasmid used: the luciferase expression 48 hours after EGT of pCMV-luc and the frequency of the IFNγ+ hTERT-specific CD8 T-cells 14 days after EGT of INVAC-1. Both the luciferase expression (Figure 1a) and the frequency of IFNγ+ hTERT-specific CD8 T-cells (Figure 1b) were significantly increased when EP were applied directly after ID DNA injection into the dermis (P < 0.01 and P < 0.05, respectively), in comparison with animals which received ID DNA injection without EP. Thus, electrotransfer is essential to reach significant levels of luciferase expression after ID injection of pCMV-luc and to induce significant levels of hTERT-specific CD8 T-cell responses after immunization with an ID injection of INVAC-1.


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)

Comparison of pCMV-luc gene transfer into the dermis and INVAC-1-mediated ID vaccination efficiencies with or without EP using plate electrodes. (a) Representation of bioluminescence intensities in C57BL/6J mice 2 days after pCMV-luc ID injection followed or not by EP, n = 5 mice for pCMV-luc ID injection alone, n = 10 (from 5 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with 25 µg of INVAC-1 followed or not by EP, n = 8 both for INVAC-1 ID injection alone or n = 6 for INVAC-1 ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, Mann–Whitney–Wilcoxon test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Comparison of pCMV-luc gene transfer into the dermis and INVAC-1-mediated ID vaccination efficiencies with or without EP using plate electrodes. (a) Representation of bioluminescence intensities in C57BL/6J mice 2 days after pCMV-luc ID injection followed or not by EP, n = 5 mice for pCMV-luc ID injection alone, n = 10 (from 5 mice, 2 treatments per mouse) for pCMV-luc ID injection+EP. (b) Frequency of hTERT-specific IFNγ+ CD8 T-cells detected in C57BL/6J mice vaccinated 14 days before with 25 µg of INVAC-1 followed or not by EP, n = 8 both for INVAC-1 ID injection alone or n = 6 for INVAC-1 ID injection+EP. Bars represent median values, *P < 0.05, **P < 0.01, Mann–Whitney–Wilcoxon test.
Mentions: C57BL/6J mice were injected intradermally on both flanks with either pCMV-luc (encoding luciferase) or INVAC-1 (encoding a modified form of hTERT) plasmids, followed or not by the application of EP (1 HV pulse of 1,000 V/cm and of 100 µs followed 1,000 ms later by 1 LV pulse of 140 V/cm and of 400 ms).1 Erythema was observed neither after mice shaving nor during or after the EGT procedure. Two parameters were measured after EGT according to the plasmid used: the luciferase expression 48 hours after EGT of pCMV-luc and the frequency of the IFNγ+ hTERT-specific CD8 T-cells 14 days after EGT of INVAC-1. Both the luciferase expression (Figure 1a) and the frequency of IFNγ+ hTERT-specific CD8 T-cells (Figure 1b) were significantly increased when EP were applied directly after ID DNA injection into the dermis (P < 0.01 and P < 0.05, respectively), in comparison with animals which received ID DNA injection without EP. Thus, electrotransfer is essential to reach significant levels of luciferase expression after ID injection of pCMV-luc and to induce significant levels of hTERT-specific CD8 T-cell responses after immunization with an ID injection of INVAC-1.

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