title: Liposome-mediated gene transfer in rat lung transplantation: A comparison between the in vivo and ex vivo approaches.
abstract: OBJECTIVE
  We compared the efficacy of in vivo and ex vivo liposome transfection in rat lung transplantation.
  
  
  METHODS
  (1) Chloramphenicol acetyltransferase group: Fischer rats underwent isogeneic transplantation (n = 4 per group). Recipients were put to death on postoperative day 2 for chloramphenicol acetyltransferase activity. Ex vivo setting: Grafts received cDNA complexed or not with liposomes and were transplanted after 1.5 or 10 hours at 10 degreesC. In vivo setting: Donors were intravenously injected with cDNA complexed or not with liposomes. Lungs were harvested after 1.5 or 10 hours, preserved at 10 degreesC, and transplanted. (2) Transforming growth factor-beta1 group: Brown-Norway rats served as donors and Fischer rats as recipients. All grafts were preserved for 3 hours at 10 degreesC. On postoperative day 5, arterial oxygenation and histologic rejection scores were assessed. Ex vivo setting: Grafts received transforming growth factor-beta1 sense (n = 8) or antisense (n = 7) complexed with liposomes or cDNA alone (n = 5). In vivo setting: Donors were intravenously injected with liposome:transforming growth factor-beta1 sense cDNA (n = 7). Exposure time was 3 hours.
  
  
  RESULTS
  (1) Chloramphenicol acetyltransferase-transfection was superior in the ex vivo group but was not statistically different for longer exposure times. (2) Transforming growth factor-beta1-arterial oxygenation was superior in the ex vivo liposome:sense group. cDNA alone was inefficient. Rejection scores were not statistically different between ex vivo and in vivo liposome:sense groups but were better when the ex vivo liposome:sense group was compared with the cDNA alone or the antisense groups.
  
  
  CONCLUSIONS
  (1) With current liposome technology, the ex vivo route is superior to the in vivo approach; (2) cDNA alone does not provide transgene expression at levels to produce a functional effect.
authors.affiliation: Division of Cardiothoracic Surgery, Department of Surgery, and Department of Pathology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: C H
authors.initials: CH
authors.lastname: Boasquevisque
authors.forename: B N
authors.initials: BN
authors.lastname: Mora
authors.forename: M
authors.initials: M
authors.lastname: Boglione
authors.forename: J K
authors.initials: JK
authors.lastname: Ritter
authors.forename: R K
authors.initials: RK
authors.lastname: Scheule
authors.forename: N
authors.initials: N
authors.lastname: Yew
authors.forename: L
authors.initials: L
authors.lastname: Debruyne
authors.forename: L
authors.initials: L
authors.lastname: Qin
authors.forename: J S
authors.initials: JS
authors.lastname: Bromberg
authors.forename: G A
authors.initials: GA
authors.lastname: Patterson
mesh_terms: D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D018076:DNA, Complementary; D015870:Gene Expression; D005796:Genes; D017930:Genes, Reporter; D008081:Liposomes; D008168:Lung / Q000201:enzymology; D016040:Lung Transplantation / Q000473:pathology / Q000502:physiology; D008297:Male; D051381:Rats; D011914:Rats, Inbred BN; D011916:Rats, Inbred F344; D014162:Transfection / Q000379:methods*; D016212:Transforming Growth Factor beta / Q000235:genetics; D019076:Transgenes; D014185:Transplantation, Isogeneic
chemical_list: D018076:DNA, Complementary; D008081:Liposomes; D016212:Transforming Growth Factor beta; D015500:Chloramphenicol O-Acetyltransferase