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.
Division of Thoracic and Cardiovascular Surgery, Niigata University Graduate School of Medicine and Dental Science, Niigata, Japan. kentsuchi@hotmail.com
Masanori
M
Tsuchida
Terumoto
T
Koike
Masaru
M
Takekubo
Hiroyuki
H
Hirahara
Haruo
H
Hanawa
Hiroki
H
Maruyama
Jun-ichi
J
Miyazaki
Jun-ichi
J
Hayashi
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