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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

INVAC-1 plasmid map. Bases 1–3478: NTC8685-eRNA41H-HindIII-XbaI vector (NTC); Bases 3479–3484: HindIII cloning site (NTC/Invectys); Bases 3485–6967: Ubi-Telomerase transgene (Invectys); Bases 6968–6973: XbaI cloning site (Invectys/NTC); Bases 6974–7120: NTC8685-eRNA41H-HindIII-XbaI vector (NTC).
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fig7: INVAC-1 plasmid map. Bases 1–3478: NTC8685-eRNA41H-HindIII-XbaI vector (NTC); Bases 3479–3484: HindIII cloning site (NTC/Invectys); Bases 3485–6967: Ubi-Telomerase transgene (Invectys); Bases 6968–6973: XbaI cloning site (Invectys/NTC); Bases 6974–7120: NTC8685-eRNA41H-HindIII-XbaI vector (NTC).

Mentions: INVAC-1 is a double stranded plasmid DNA of 7,120 bp encoding a modified sequence of the telomerase protein fused to the Ubiquitin protein sequence. The encoded telomerase protein is enzymatically inactive, due to the deletion of three amino acids inside the catalytic site of hTERT (delta VDD), but can still induce immune responses against telomerase epitopes in vivo. The ubiquitin-telomerase insert is cloned into the NTC8685-ERNA41H-HindIII-XbaI expression vector designed by Nature Technology Corporation (Lincoln, Nebraska). The presence of the ubiquitin increases the addressing of the TERT protein to the proteasome and increases the MHC class I presentation pathway of TERT-derived peptides.33,34 The DNA sequence coding for the TERT protein was deleted of 47 amino acids in the N-terminal region, which encodes the nucleolar localization signal. Figure 7 represents INVAC-1 plasmid map.


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)

INVAC-1 plasmid map. Bases 1–3478: NTC8685-eRNA41H-HindIII-XbaI vector (NTC); Bases 3479–3484: HindIII cloning site (NTC/Invectys); Bases 3485–6967: Ubi-Telomerase transgene (Invectys); Bases 6968–6973: XbaI cloning site (Invectys/NTC); Bases 6974–7120: NTC8685-eRNA41H-HindIII-XbaI vector (NTC).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig7: INVAC-1 plasmid map. Bases 1–3478: NTC8685-eRNA41H-HindIII-XbaI vector (NTC); Bases 3479–3484: HindIII cloning site (NTC/Invectys); Bases 3485–6967: Ubi-Telomerase transgene (Invectys); Bases 6968–6973: XbaI cloning site (Invectys/NTC); Bases 6974–7120: NTC8685-eRNA41H-HindIII-XbaI vector (NTC).
Mentions: INVAC-1 is a double stranded plasmid DNA of 7,120 bp encoding a modified sequence of the telomerase protein fused to the Ubiquitin protein sequence. The encoded telomerase protein is enzymatically inactive, due to the deletion of three amino acids inside the catalytic site of hTERT (delta VDD), but can still induce immune responses against telomerase epitopes in vivo. The ubiquitin-telomerase insert is cloned into the NTC8685-ERNA41H-HindIII-XbaI expression vector designed by Nature Technology Corporation (Lincoln, Nebraska). The presence of the ubiquitin increases the addressing of the TERT protein to the proteasome and increases the MHC class I presentation pathway of TERT-derived peptides.33,34 The DNA sequence coding for the TERT protein was deleted of 47 amino acids in the N-terminal region, which encodes the nucleolar localization signal. Figure 7 represents INVAC-1 plasmid map.

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