Low-dose endobronchial gene transfer to ameliorate lung graft ischemia-reperfusion injury.
OBJECTIVE
This study was undertaken to determine whether low-dose endobronchial transfer to the donor of the gene for human interleukin 10 would decrease ischemia-reperfusion injury in lung transplantation.


METHODS
Experiments used male Fischer rats. Donor animals underwent right thoracotomy. A catheter was introduced into the left main bronchus, and vector was instilled. Group I (n = 6) received 2 x 10(7) plaque-forming units of adenovirus encoding human interleukin 10, group II (n = 6) received an adenovirus control encoding beta-galactosidase, and group III (n = 6) received saline solution. After instillation the left main bronchus was clamped for 60 minutes. Lungs were removed 24 hours later and stored in low-potassium dextran glucose solution for 18 hours before left lung transplantation. Graft function was assessed at 24 hours immediately before the animals were killed. Ratio of wet to dry weight and tissue myeloperoxidase activity were measured. Transgenic expression of human interleukin 10 was evaluated by means of enzyme-linked immunosorbent assay and immunohistochemical assay.


RESULTS
Arterial oxygenation was significantly improved in group I relative to groups II and III (257.6 +/- 59.7 mm Hg vs 114.6 +/- 66.9 mm Hg and 118.6 +/- 91.1 mm Hg, P =.008 and P =.007, respectively). Neutrophil sequestration, as measured by myeloperoxidase activity, was also significantly reduced in group I relative to groups II and III (0.141 +/- 0.025 vs 0.304 +/- 0.130 and 0.367 +/- 0.153 Delta optical density units/[min. mg protein], P =.029 and P =.004, respectively). Enzyme-linked immunosorbent assay and immunohistochemical assay demonstrated the expression of human interleukin 10 in transfected lungs only.


CONCLUSIONS
Low-dose endobronchial transfer to the donor of the gene for human interleukin 10 ameliorated ischemia-reperfusion injury in rodent lung transplantation by improving graft oxygenation and reducing neutrophil sequestration. Only 2 x 10(7) plaque-forming units of adenoviral vector were required for functional transgenic expression. Endobronchial gene transfer to lung grafts may be a useful delivery route even at low doses.
Tagawa
Tsutomu
T
Division of Cardiothoracic Surgery and the Department of Surgery, Washington University School of Medicine, Barnes-Jewish Hospital, St Louis, MO 63110-1013, USA.
Suda
Takashi
T
Daddi
Niccolò
N
Kozower
Benjamin D
BD
Kanaan
Samer A
SA
Mohanakumar
T
T
Patterson
G Alexander
GA
D000818:Animals; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D007150:Immunohistochemistry; D016753:Interleukin-10 / Q000096:biosynthesis / Q000235:genetics; D008168:Lung / Q000378:metabolism*; D016040:Lung Transplantation; D008297:Male; D009195:Peroxidase / Q000378:metabolism; D011659:Pulmonary Gas Exchange / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344; D015427:Reperfusion Injury / Q000209:etiology* / Q000517:prevention & control*
D016753:Interleukin-10; D009195:Peroxidase