title: Transfection efficiency and toxicity following delivery of naked plasmid DNA and cationic lipid-DNA complexes to ovine lung segments.
abstract: We defined, using a novel large animal model system, the acute pathologic response to localized pulmonary administration of either naked plasmid DNA (pDNA) or cationic lipid-pDNA complexes (pDNA:GL67) and related such responses to concomitant indicators of transfection efficiency, namely levels of chloramphenicol acetyl transferase (CAT) protein and mRNA in specific lung tissue compartments. We instilled doses of 0.2, 1, and 5 mg pDNA to spatially distinct lung segments in six anesthetized sheep and doses of 0.2, 1, and 5 mg pDNA:GL67 to a further six sheep. Twenty-four hours after gene delivery the sheep were euthanized and necropsy examination with sampling of relevant tissues was carried out. Levels of plasmid-derived CAT-specific mRNA and CAT protein in samples derived from segments treated with either pDNA or pDNA:GL67 increased in relation to the administered dose. Levels of mRNA and protein expression were greater for pDNA:GL67 than for pDNA alone. A significant correlation was observed between mRNA and protein expression in samples derived from airways treated with pDNA:GL67. Histopathological changes following administration of both pDNA and pDNA:GL67 were characterized by a neutrophilic inflammation predominantly oriented on airways. The severity of the inflammatory response appeared to correlate with the administered dose of DNA and was generally more severe for pDNA:GL67.
authors.affiliation: Medical Genetics Section, School of Molecular & Clinical Medicine, and Department of Veterinary Pathology, College of Medicine and Veterinary Medicine, University of Edinburg, Edinburgh EH8 9AG, United Kingdom.
authors.forename: Michael
authors.initials: M
authors.lastname: Emerson
authors.forename: Louise
authors.initials: L
authors.lastname: Renwick
authors.forename: Stephen
authors.initials: S
authors.lastname: Tate
authors.forename: Susan
authors.initials: S
authors.lastname: Rhind
authors.forename: Elspeth
authors.initials: E
authors.lastname: Milne
authors.forename: Hazel Alsop
authors.initials: HA
authors.lastname: Painter
authors.forename: A Christopher
authors.initials: AC
authors.lastname: Boyd
authors.forename: Gerry
authors.initials: G
authors.lastname: McLachlan
authors.forename: Uta
authors.initials: U
authors.lastname: Griesenbach
authors.forename: Seng H
authors.initials: SH
authors.lastname: Cheng
authors.forename: Deborah R
authors.initials: DR
authors.lastname: Gill
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Hyde
authors.forename: Alison
authors.initials: A
authors.lastname: Baker
authors.forename: Eric W
authors.initials: EW
authors.lastname: Alton
authors.forename: David J
authors.initials: DJ
authors.lastname: Porteous
authors.forename: D David S
authors.initials: DD
authors.lastname: Collie
mesh_terms: D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D008055:Lipids / Q000494:pharmacology* / Q000633:toxicity; D008168:Lung / Q000187:drug effects* / Q000473:pathology; D010957:Plasmids / Q000494:pharmacology* / Q000633:toxicity; D012756:Sheep / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D008055:Lipids; D015500:Chloramphenicol O-Acetyltransferase