Exosurf enhances adenovirus-mediated gene transfer to alveolar type II cells.
We assessed the role of surfactant replacement mixtures in the enhancement of adenovirus-mediated gene transfer to pulmonary epithelial cells both in vitro and in vivo. A549 cells, a pulmonary epithelium-derived adenocarcinoma cell line, were incubated with either media alone or media containing 10 microg phospholipid/ml Exosurf or Infasurf for 50 min followed by addition of a replication-deficient adenovirus (E1-deleted) expressing the luciferase reporter gene [AdCMV-Luc; 10 plaque-forming units (PFU)/cell] for 4 h. Pretreatment with Exosurf, but not Infasurf, at 37 degrees C, but not at 4 degrees C, enhanced luciferase activity in A549 cells 24 h later by 156% (P < 0.01). Intratracheal instillation of AdCMV-Luc (2 x 10(9) PFU) into rats resulted in luciferase expression mainly in alveolar macrophages and to a smaller extent in alveolar type II (ATII) cells 24 h later. However, when the AdCMV-Luc instillation was preceded by Exosurf (250 microl; 25 mg/ml), a 10-fold increase in ATII cell luciferase activity was noted. Preincubation of cultured ATII cells with Exosurf also enhanced their transfection by AdCMV-Luc by 515% (P < 0.001). The results of these studies provide a new strategy for targeting ATII cells for gene delivery.
Manuel
S M
SM
Department of Anesthesiology, University of Alabama at Birmingham, 35233-6810, USA.
Guo
Y
Y
Matalon
S
S
D000230:Adenocarcinoma; D000256:Adenoviridae; D000818:Animals; D002478:Cells, Cultured; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005233:Fatty Alcohols / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D008175:Lung Neoplasms; D008297:Male; D010767:Phosphorylcholine; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D005233:Fatty Alcohols; D011663:Pulmonary Surfactants; D010767:Phosphorylcholine; D011092:Polyethylene Glycols; C047049:dipalmitoylphosphatidylcholine, hexadecanol, tyloxapol drug combination; D008156:Luciferases