Electroporation-mediated transfer of plasmid DNA encoding IL-10 attenuates orthotopic tracheal allograft stenosis in rats.
BACKGROUND
Electroporation has been shown to increase the efficacy of intramuscular injection of plasmid DNA, resulting in a higher level of foreign gene expression. Using this technique, we examined the effect of viral IL-10 gene transfer on the prevention of tracheal allograft stenosis in an animal model.


METHODS
On the day of tracheal transplantation, recipient Lewis rats were intramuscularly injected with either plasmid pCAGGS-LacZ or plasmid pCAGGS-viral IL-10, followed immediately by electroporation. Tracheas from Brown Norway donors were transplanted into the backs of Lewis recipients, and the histology of the grafts were assessed 2 and 4 weeks after transplantation.


RESULTS
The serum level of IL-10 peaked at 2000 pg/ml one day after injection; the level then slowly decreased, but was maintained above 1000 pg/ml until 8 days after injection. At Day 28, the airway lumina of the tracheal allografts were almost completely obliterated by fibroproliferative tissue in the control pCAGGS-LacZ-treated rats. In rats injected once with pCAGGS-viral IL-10, luminal obliteration was significantly decreased compared with the control pCAGGS-LacZ-treated rats (mean luminal opening 46.8% vs 0% p<0.05). The loss of epithelial cells lining the airway was also decreased in the IL-10-treated group (mean epithelial coverage 42% vs 5% p<0.05). Multiple injections with pCAGGS-viral IL-10 did not further improve the histological changes.


CONCLUSIONS
IL-10 gene transfer by intramuscular injection using electroporation attenuated tracheal allograft stenosis associated with mild epithelial injury.
Tsuchida
Masanori
M
Division of Thoracic and Cardiovascular Surgery, Niigata University Graduate School of Medicine and Dental Science, Niigata, Japan. kentsuchi@hotmail.com
Koike
Terumoto
T
Takekubo
Masaru
M
Hirahara
Hiroyuki
H
Hanawa
Haruo
H
Maruyama
Hiroki
H
Miyazaki
Jun-ichi
J
Hayashi
Jun-ichi
J
D000818:Animals; D001989:Bronchiolitis Obliterans / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D004269:DNA, Bacterial; D004279:DNA, Viral; D018274:Electroporation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007273:Injections, Intramuscular; D016753:Interleukin-10 / Q000235:genetics* / Q000276:immunology / Q000627:therapeutic use; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D014132:Trachea / Q000276:immunology / Q000473:pathology / Q000637:transplantation*; D014135:Tracheal Stenosis / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D019076:Transgenes / Q000276:immunology; D014184:Transplantation, Homologous / Q000276:immunology
D004269:DNA, Bacterial; D004279:DNA, Viral; D016753:Interleukin-10