Enhanced distribution of adenovirus-mediated gene transfer to lung parenchyma by perfluorochemical liquid.
Although gene therapy holds great promise for the treatment of inherited and acquired diseases of the lung, a number of issues including efficient delivery and distribution of genes to pulmonary target cells must still be addressed. In this study we evaluated the use of perfluorochemical (PFC) liquid as a vehicle for delivery of recombinant adenovirus (AdCBlacZ) to lungs of juvenile rabbits. Virus was instilled into trachea of rabbits, and 4 days later the lungs were removed, cut into multiple pieces, and assayed for beta-galactosidase (beta-Gal) activity. Total lung expression of the beta-Gal reporter gene was increased two- to three-fold by instillation of the virus (10(11) particles/kg body weight) in saline (1.5 ml/kg) simultaneously with perflubron liquid (15 ml/kg) compared to virus+saline alone (control). Uniformity of beta-Gal activity between lobes was significantly improved by the PFC liquid. In perflubron-treated lungs approximately 45% of the lung pieces had beta-Gal-specific activity values within 50-150% of the mean specific activity for the total lung, compared to only approximately 15% of the pieces in control lungs. More of total lobar beta-Gal activity was recovered in the distal lung tissue (approximately two-fold greater than controls, p < 0.05). Morphological assessment of X-Gal-stained, fresh-frozen lung sections showed increased levels and more complete staining of alveolar wall cells in the PFC group. These data indicate that the PFC liquid perflubron enhances distribution of virus-mediated gene expression to the lung parenchyma in healthy rabbits. PFC liquid may be a useful treatment vehicle for accessing distal spaces of the damaged or diseased lung.
Lisby
D A
DA
Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Ballard
P L
PL
Fox
W W
WW
Wolfson
M R
MR
Shaffer
T H
TH
Gonzales
L W
LW
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001835:Body Weight / Q000502:physiology; D004274:DNA, Recombinant / Q000235:genetics; D005240:Feasibility Studies; D005466:Fluorocarbons; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008168:Lung / Q000000981:diagnostic imaging / Q000502:physiology*; D009929:Organ Size / Q000502:physiology; D014677:Pharmaceutical Vehicles; D011817:Rabbits; D011859:Radiography; D001616:beta-Galactosidase / Q000235:genetics
D004274:DNA, Recombinant; D005466:Fluorocarbons; D014677:Pharmaceutical Vehicles; D001616:beta-Galactosidase