title: Surfactant protein A-polylysine conjugates for delivery of DNA to airway cells in culture.
abstract: Surfactant protein A (SP-A) was modified by covalent linkage with polylysine of average M(r) 21 kD ([Lys]21kD-SP-A) and utilized to transfect human airway epithelial cells (H441) in vitro. Transfection of H441 cells was more efficient with [Lys]21kD-SP-A than with polylysine-DNA or unmodified SP-A-DNA complexes. Transfection with [Lys]21kD-SP-A was effective at a protein-to-DNA molar ratio of 400:1 and in the presence of an exogenous surfactant preparation, Survanta. Transfection with [Lys]21kD-SP-A was reduced in the presence of unmodified SP-A consistent with the concept of a receptor mediated uptake of protein-DNA complexes. Increased transfection efficiency correlated with decreasing diameter of the [Lys]21kD-SP-A-DNA complexes, and these complexes bound to the cell surface and pseudopodia of H441 cells. Transfection was enhanced by co-incubation with replication-deficient adenovirus. Cotransfection by [Lys]21kD-SP-A-DNA and [Lys]10kD-SP-B resulted in an additive level of reporter gene (CAT) expression. [Lys]21kD-SP-A-DNA is likely to be useful as a component of a surfactant-based DNA delivery system for transfection of airway cells.
authors.affiliation: Division of Pulmonary Biology, Children's Hospital Medical Center, Cincinnati OH 45229-3039, USA.
authors.forename: G F
authors.initials: GF
authors.lastname: Ross
authors.forename: R E
authors.initials: RE
authors.lastname: Morris
authors.forename: G
authors.initials: G
authors.lastname: Ciraolo
authors.forename: K
authors.initials: K
authors.lastname: Huelsman
authors.forename: M
authors.initials: M
authors.lastname: Bruno
authors.forename: J A
authors.initials: JA
authors.lastname: Whitsett
authors.forename: J E
authors.initials: JE
authors.lastname: Baatz
authors.forename: T R
authors.initials: TR
authors.lastname: Korfhagen
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D001688:Biological Products; D004247:DNA / Q000235:genetics / Q000648:ultrastructure; D008168:Lung / Q000166:cytology*; D008854:Microscopy, Electron; D011107:Polylysine / Q000737:chemistry; D011510:Proteolipids / Q000737:chemistry / Q000648:ultrastructure; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D001688:Biological Products; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D011107:Polylysine; D004247:DNA; C072197:beractant