Adenovirus mediated gene transfer into rat lung grafts at the time of harvest.
OBJECTIVE
New methods to introduce genetic material into cells in vivo may revolutionize current treatment modalities. Expression of functional genes in lung allografts could be used as a prophylactic strategy for reperfusion injury and rejection. We studied the feasibility of ex vivo adenovirus mediated transfection of rat lung allografts.


METHODS
In group I (n = 3) donor rat lungs (Fisher) were flushed with Low Potassium Dextran Glucose (LPDG) solution (20 ml, 20 degrees C). 4 x 10(11) viral particles of adenovirus 5 containing the E. coli lacZ reporter gene coding for beta-galactosidase (AdCMV-beta-Gal) were added to the last milliliter of the flush solution. Lung grafts were stored for 3.5 h at room temperature followed by syngenic orthotopic transplantation (Fisher to Fisher) using a microsurgical cuff technique. On postoperative day 5 the heart lung block was extracted and flushed with x-Gal (beta-Gal substrate) and kept in x-Gal for 3 h at 37 degrees C. Color development was observed macroscopically and plastic embedded sections were used for histologic examination. Group II grafts (n = 3) served as controls and were flushed without adenovirus.


RESULTS
X-Gal stained the transfected lung grafts blue, indicating high reporter gene expression. Control lungs did not stain with x-Gal. In group I histological examination demonstrated transfection predominantly in type II pneumocytes. Surprisingly endothelial cells showed no beta-Gal activity.


CONCLUSIONS
This study demonstrates that ex vivo transfection of lung grafts at the time of harvest is a feasible method of gene transfer and results in gene expression after transplantation.
Schmid
R A
RA
Department of Surgery, Barnes Hospital, Washington University School of Medicine, St. Louis, MO 63110, USA.
Narita
M
M
Boasquevisque
C H
CH
Ando
K
K
Botney
M D
MD
Cooper
J D
JD
Schwartz
A L
AL
Patterson
G A
GA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology; D005240:Feasibility Studies; D015870:Gene Expression; D018014:Gene Transfer Techniques; D016040:Lung Transplantation / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011916:Rats, Inbred F344; D009927:Tissue and Organ Procurement; D014184:Transplantation, Homologous; D001616:beta-Galactosidase / Q000235:genetics
D001426:Bacterial Proteins; D001616:beta-Galactosidase