title: Feline immunodeficiency virus and retrovirus-mediated adventitial ex vivo gene transfer to rabbit carotid artery using autologous vascular smooth muscle cells.
abstract: We have developed an ex vivo gene transfer technique to rabbit arterial wall using autologous smooth muscle cells (SMCs). SMCs were harvested from rabbit ear artery, transduced in vitro with vesicular stomatitis virus G-glycoprotein pseudotyped retrovirus or feline immunodeficiency virus (FIV) and returned to the adventitial surface of the carotid artery using a periadventitial silicone collar or collagen sheet placed around the artery. Beta-galactosidase (lacZ) and human apolipoprotein E3 (apoE3) cDNAs were used as transgenes. After retrovirus-mediated gene transfer of lacZ the selected cells implanted with high efficiency and expressed lacZ marker gene at a very high level 7 and 14 days after the operation. The level of lacZ expression decreased thereafter but was still detectable 12 weeks after the gene transfer, and was exclusively localized to the site of cell implantation inside the collar. Utilizing FIV vector expressing apoE3, low levels of apoE were measured from serum collected from a low-density lipoprotein receptor deficient Watanabe heritable hyperlipidemic rabbits 1 month after the gene transfer. The physiological effect of apoE expression was detected as transiently elevated serum cholesterol levels. The results indicate that the model can be used for high efficiency local gene transfer in arteries, e.g. during vascular surgery. The model is also valuable for studying expression, stability and safety of new gene transfer vectors and their expression products in vivo.
authors.affiliation: Department of Biotechnology and Molecular Medicine, A I Virtanen Institute for Molecular Sciences, University of Kuopio, PO Box 1627, FIN-70211 Kuopio, Finland.
authors.forename: Hanna M
authors.initials: HM
authors.lastname: Kankkonen
authors.forename: Mikko P
authors.initials: MP
authors.lastname: Turunen
authors.forename: Mikko O
authors.initials: MO
authors.lastname: Hiltunen
authors.forename: Pauliina
authors.initials: P
authors.lastname: Lehtolainen
authors.forename: Jonna
authors.initials: J
authors.lastname: Koponen
authors.forename: Pia
authors.initials: P
authors.lastname: Leppänen
authors.forename: Anna-Mari
authors.initials: AM
authors.lastname: Turunen
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
mesh_terms: D000818:Animals; D053318:Apolipoprotein E3; D001057:Apolipoproteins E / Q000235:genetics; D002339:Carotid Arteries / Q000166:cytology / Q000502:physiology; D002415:Cats; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D016089:Immunodeficiency Virus, Feline; D008562:Membrane Glycoproteins / Q000235:genetics; D032389:Myocytes, Smooth Muscle / Q000502:physiology* / Q000637:transplantation; D011817:Rabbits; D014161:Transduction, Genetic / Q000379:methods; D014182:Transplantation, Autologous; D014652:Vascular Diseases / Q000628:therapy; D014656:Vascular Surgical Procedures; D014759:Viral Envelope Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D053318:Apolipoprotein E3; D001057:Apolipoproteins E; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D001616:beta-Galactosidase