Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs.
BACKGROUND
Experiments were designed to study the efficiency, distribution, and toxicity of transbronchial adenoviral-mediated transfer of endothelial constitutive nitric oxide synthase (ecNOS) gene to transplanted lungs.


METHODS
Syngeneic orthotopic single-lung transplantation in the rat was performed after airway administration (300 microL, 1 x 10(9) pfu/mL) of either the ecNOS gene or the marker gene beta-Gal (control group) to donor lungs (n=4 each). After 4 days, transgene expression, inflammation, and the presence of apoptosis in the transplanted lungs were assessed by molecular, immunohistochemical, and histologic techniques.


RESULTS
Gene transfer was confirmed by a positive polymerase chain reaction signal for the recombinant ecNOS gene, and recombinant messenger RNA by reverse transcription polymerase chain reaction. Positive immunohistochemical staining for ecNOS was present in more than 75% of pneumocytes only in ecNOS transduced lungs. Calcium-dependent nitric oxide synthase activity was increased in ecNOS- compared with betaGal-transduced lungs (2,139+/-756 versus 47+/-28 pmol x mg protein(-1) x h(-1); p < 0.05). Minimal to mild inflammation was observed in all transplanted lungs; fewer than 0.5% of cells in both groups were apoptotic.


CONCLUSIONS
Transbronchial transfer of ecNOS gene to the transplanted lung results in transduction of pneumocytes with expression of a functionally active transgene product.
Jeppsson
A
A
Department of Surgery, Mayo Clinic and Foundation, Rochester, Minnesota 55905, USA.
Pellegrini
C
C
O'Brien
T
T
Miller
V M
VM
Tazelaar
H D
HD
McGregor
C G
CG
D000256:Adenoviridae; D000818:Animals; D017209:Apoptosis; D001980:Bronchi; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D007150:Immunohistochemistry; D008168:Lung / Q000201:enzymology* / Q000473:pathology; D016040:Lung Transplantation; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III; D016133:Polymerase Chain Reaction; D051381:Rats; D011917:Rats, Inbred Lew; D019076:Transgenes; D001616:beta-Galactosidase / Q000235:genetics
D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D001616:beta-Galactosidase