Successful in vivo and ex vivo transfection of pulmonary artery segments in lung isografts.
OBJECTIVE
Gene transfer to lung grafts may be useful in ameliorating ischemia-reperfusion injury and rejection. Efficient gene transfection to the whole organ may prove problematic. Proximal pulmonary artery endothelial transfection might provide beneficial downstream effects on the whole graft. The aim of this study was to determine the feasibility of transfecting proximal pulmonary artery segments in lung isografts.


METHODS
Male Fischer rats were divided into six groups. In vivo transfection: In group I (n = 7), a proximal segment of the left pulmonary artery was isolated and injected with saline solution by means of a catheter inserted through the right ventricle. After an exposure period of 20 minutes, clamps were removed and blood flow was restored. In group II (n = 7), the isolated arterial segments were injected with adenovirus carrying the Escherichia coli LacZ gene encoding for beta-galactosidase. Ex vivo transfection: In group III (n = 5), arterial segments were injected ex vivo with saline solution and in group IV (n = 5) with the adenovirus construct. In group V (n = 6), arteries were injected with saline solution and in group VI (n = 11) with liposome chloramphenicol acetyl transferase cDNA. In groups I to IV, animals were killed on postoperative day 3 and transgene expression was assessed by Bluo-Gal staining. In groups V and VI, animals were killed on postoperative day 2 and transgene expression was assessed by chloramphenicol acetyl transferase activity assay.


RESULTS
Transgene expression was detected grossly and microscopically in endothelial and smooth muscle cells of pulmonary artery segments from all surviving animals of groups II and IV. In group VI, chloramphenicol acetyl transferase activity was significant in all assessed arterial segments.


CONCLUSIONS
Significant transgene expression is observed in proximal pulmonary artery segments after both in vivo and ex vivo exposure.
Yano
M
M
Division of Cardiothoracic Surgery, Washington University School of Medicine, St. Louis, Mo., USA.
Boasquevisque
C H
CH
Scheule
R K
RK
Botney
M D
MD
Cooper
J D
JD
Patterson
G A
GA
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000008:administration & dosage; D004730:Endothelium, Vascular; D005240:Feasibility Studies; D005822:Genetic Vectors; D006084:Graft Rejection / Q000517:prevention & control; D008081:Liposomes; D016040:Lung Transplantation / Q000502:physiology; D008297:Male; D011651:Pulmonary Artery / Q000201:enzymology; D051381:Rats; D011916:Rats, Inbred F344; D015427:Reperfusion Injury / Q000517:prevention & control; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014185:Transplantation, Isogeneic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; D008081:Liposomes; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase