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Gene expression and immune response kinetics using electroporation-mediated DNA delivery to muscle

Gronevik, E; Vult von Steyern, Fredrik LU ; Kalhovde, JM; Tjelle, TE and Mathiesen, I (2005) In Journal of Gene Medicine 7(2). p.218-227
Abstract
Background Injection of DNA encoding exogenic proteins into muscle tissue combined with electroporation often results in a transient increase of the encoded protein concentration in the muscle and the blood. The reduction is normally due to an immune response against the exogenic protein but other factors may also be involved. How various electroporation parameters affect the concentration kinetics of syngenic and exogenic proteins is studied in relation to immune response and muscle damage after electroporation-mediated DNA transfer to muscle. Methods Electroporation was applied to mouse quadriceps muscles after injection of DNA encoding either secreted alkaline phosphatase (SEAP) or a mouse IgG molecule. Protein concentrations in blood... (More)
Background Injection of DNA encoding exogenic proteins into muscle tissue combined with electroporation often results in a transient increase of the encoded protein concentration in the muscle and the blood. The reduction is normally due to an immune response against the exogenic protein but other factors may also be involved. How various electroporation parameters affect the concentration kinetics of syngenic and exogenic proteins is studied in relation to immune response and muscle damage after electroporation-mediated DNA transfer to muscle. Methods Electroporation was applied to mouse quadriceps muscles after injection of DNA encoding either secreted alkaline phosphatase (SEAP) or a mouse IgG molecule. Protein concentrations in blood or muscle and antibody responses were measured for a period up to 3 months. Tissue inflammation and muscle cell damage were studied on muscle cross-sections and assessed by measuring the concentrations of creatine phosphokinase (CPK) in blood. Results Mice with the highest SEAP concentration in blood at day 7 also had the highest rate of decrease afterwards, the strongest antibody responses against SEAP and the highest acute levels of CPK in blood. DNA-transfected muscle fibers were significantly reduced in number from days 7 to 14. Mononuclear cells surrounded the reporter gene expressing muscle fibers, thus indicating a cellular immune response. When using DNA encoding a syngenic protein the protein concentration in blood was relatively stabile over a 3-month period, but showed different kinetics for various electroporation parameters. Conclusions Our findings suggest that the optimal electroporation. parameters for DNA vaccination may be different from the optimal parameters for long-term expression of genes encoding syngenic proteins. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
transfection, intramuscular injection, electroporation, electroporation parameters, gene therapy, DNA vaccination
in
Journal of Gene Medicine
volume
7
issue
2
pages
218 - 227
publisher
John Wiley & Sons
external identifiers
  • wos:000227253900009
  • pmid:15515140
  • scopus:14844291308
ISSN
1521-2254
DOI
10.1002/jgm.650
language
English
LU publication?
yes
id
91f5a353-bd90-4d86-af1d-effa198e13d5 (old id 251809)
date added to LUP
2007-08-14 09:07:06
date last changed
2017-01-15 03:35:22
@article{91f5a353-bd90-4d86-af1d-effa198e13d5,
  abstract     = {Background Injection of DNA encoding exogenic proteins into muscle tissue combined with electroporation often results in a transient increase of the encoded protein concentration in the muscle and the blood. The reduction is normally due to an immune response against the exogenic protein but other factors may also be involved. How various electroporation parameters affect the concentration kinetics of syngenic and exogenic proteins is studied in relation to immune response and muscle damage after electroporation-mediated DNA transfer to muscle. Methods Electroporation was applied to mouse quadriceps muscles after injection of DNA encoding either secreted alkaline phosphatase (SEAP) or a mouse IgG molecule. Protein concentrations in blood or muscle and antibody responses were measured for a period up to 3 months. Tissue inflammation and muscle cell damage were studied on muscle cross-sections and assessed by measuring the concentrations of creatine phosphokinase (CPK) in blood. Results Mice with the highest SEAP concentration in blood at day 7 also had the highest rate of decrease afterwards, the strongest antibody responses against SEAP and the highest acute levels of CPK in blood. DNA-transfected muscle fibers were significantly reduced in number from days 7 to 14. Mononuclear cells surrounded the reporter gene expressing muscle fibers, thus indicating a cellular immune response. When using DNA encoding a syngenic protein the protein concentration in blood was relatively stabile over a 3-month period, but showed different kinetics for various electroporation parameters. Conclusions Our findings suggest that the optimal electroporation. parameters for DNA vaccination may be different from the optimal parameters for long-term expression of genes encoding syngenic proteins.},
  author       = {Gronevik, E and Vult von Steyern, Fredrik and Kalhovde, JM and Tjelle, TE and Mathiesen, I},
  issn         = {1521-2254},
  keyword      = {transfection,intramuscular injection,electroporation,electroporation parameters,gene therapy,DNA vaccination},
  language     = {eng},
  number       = {2},
  pages        = {218--227},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Gene Medicine},
  title        = {Gene expression and immune response kinetics using electroporation-mediated DNA delivery to muscle},
  url          = {http://dx.doi.org/10.1002/jgm.650},
  volume       = {7},
  year         = {2005},
}