Surfactant protein A-polylysine conjugates for delivery of DNA to airway cells in culture.
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.
Ross
G F
GF
Division of Pulmonary Biology, Children's Hospital Medical Center, Cincinnati OH 45229-3039, USA.
Morris
R E
RE
Ciraolo
G
G
Huelsman
K
K
Bruno
M
M
Whitsett
J A
JA
Baatz
J E
JE
Korfhagen
T R
TR
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
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