Gene transfer of endothelial nitric oxide synthase to pulmonary allografts: impact on acute rejection.
Experiments were designed to study whether overexpression of nitric oxide (NO) from endothelial nitric oxide synthase (eNOS) affects acute rejection. Allogenic, orthotopic single-lung transplantation was performed after transbronchial adenoviral-mediated gene transfer (3 x 10(8) pfu) of either of eNOS or beta-galactosidase to donor lungs of rats (n = 6 each). No immunosuppression was used. After 4 days, transplanted lungs were prepared for enzyme activity, cGMP and histology. Calcium-dependent NOS activity, reflecting eNOS, was greater in eNOS-transduced lungs (587 +/- 97 vs 2.1 +/- 1.4 pmol/mg protein per h, P <0.001). In contrast, calcium-independent NOS activity, reflecting iNOS, was comparable. Concentrations of cGMP were higher in eNOS-transduced lungs (13.2 +/- 2.3 vs 4.9 +/- 0.5 pmol/mg protein). Positive immunostaining for eNOS was present in pneumocytes only in eNOS-transduced lungs. No difference in histological grade of rejection was observed. eNOS gene transfer to pulmonary allografts results in a functionally active transgene product and increased NO production. Increasing NO from eNOS does not affect histogically identified acute rejection.
Jeppsson
A
A
Mayo Clinic and Foundation, Rochester, Minnesota, USA. anders.jeppsson@sahlgrenska.se
Pellegrini
C
C
O'Brien
T
T
Miller
V M
VM
Tazelaar
H D
HD
Taner
C B
CB
McGregor
C G
CG
D000256:Adenoviridae; D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D018014:Gene Transfer Techniques; D006084:Graft Rejection / Q000473:pathology*; D016040:Lung Transplantation / Q000276:immunology / Q000473:pathology / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000235:genetics* / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D014184:Transplantation, Homologous; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D000242:Cyclic AMP; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496318:Nos2 protein, rat; C496316:Nos3 protein, rat; D001616:beta-Galactosidase