Expression of human alpha 1-antitrypsin in mouse after in vivo gene transfer to hepatocytes by small liposomes.
A plasmid (pTG7101) containing the full-length human alpha 1-antitrypsin gene was encapsulated in small liposomes and used for "in vivo" gene transfer to mouse hepatocytes, by i.v. injection (100 ng DNA/mouse and dose). The expression of human protein was evaluated by microspectrophotometry after human alpha 1-antitrypsin immunoperoxidase reaction on liver cryosections and the presence in mouse plasma of de novo synthesized protein was detected by ELISA analysis. Our results indicate that a single dose of encapsulated plasmid induces the expression of human alpha 1-antitrypsin in mouse hepatocytes and a large effect (70%) remains two weeks after treatment. However, no effect was observed when mice were treated with buffer or free plasmid (100 ng/mouse) plus an equivalent lipid dose of empty liposomes. In addition, whereas no additive effect was observed after repetitive treatment-doses, the partial hepatectomy three hours after a single treatment-dose, significantly increased the presence of human alpha 1-antitrypsin in mice plasma.
Department of Pharmacology, Faculty of Medicine and Dentistry, University of Valencia, Spain.
S F
SF
Aliño
J
J
Crespo
M
M
Bobadilla
M
M
Lejarreta
C
C
Blaya
A
A
Crespo
D000818:Animals; D002478:Cells, Cultured; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D007700:Kinetics; D008081:Liposomes; D008099:Liver / Q000378:metabolism*; D051379:Mice; D010957:Plasmids; D012016:Reference Values; D013997:Time Factors; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis*
D004337:Drug Carriers; D008081:Liposomes; D000515:alpha 1-Antitrypsin