title: Adenovirus mediated gene transfer into rat lung grafts at the time of harvest.
abstract: OBJECTIVE
  New methods to introduce genetic material into cells in vivo may revolutionize current treatment modalities. Expression of functional genes in lung allografts could be used as a prophylactic strategy for reperfusion injury and rejection. We studied the feasibility of ex vivo adenovirus mediated transfection of rat lung allografts.
  
  
  METHODS
  In group I (n = 3) donor rat lungs (Fisher) were flushed with Low Potassium Dextran Glucose (LPDG) solution (20 ml, 20 degrees C). 4 x 10(11) viral particles of adenovirus 5 containing the E. coli lacZ reporter gene coding for beta-galactosidase (AdCMV-beta-Gal) were added to the last milliliter of the flush solution. Lung grafts were stored for 3.5 h at room temperature followed by syngenic orthotopic transplantation (Fisher to Fisher) using a microsurgical cuff technique. On postoperative day 5 the heart lung block was extracted and flushed with x-Gal (beta-Gal substrate) and kept in x-Gal for 3 h at 37 degrees C. Color development was observed macroscopically and plastic embedded sections were used for histologic examination. Group II grafts (n = 3) served as controls and were flushed without adenovirus.
  
  
  RESULTS
  X-Gal stained the transfected lung grafts blue, indicating high reporter gene expression. Control lungs did not stain with x-Gal. In group I histological examination demonstrated transfection predominantly in type II pneumocytes. Surprisingly endothelial cells showed no beta-Gal activity.
  
  
  CONCLUSIONS
  This study demonstrates that ex vivo transfection of lung grafts at the time of harvest is a feasible method of gene transfer and results in gene expression after transplantation.
authors.affiliation: Department of Surgery, Barnes Hospital, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Schmid
authors.forename: M
authors.initials: M
authors.lastname: Narita
authors.forename: C H
authors.initials: CH
authors.lastname: Boasquevisque
authors.forename: K
authors.initials: K
authors.lastname: Ando
authors.forename: M D
authors.initials: MD
authors.lastname: Botney
authors.forename: J D
authors.initials: JD
authors.lastname: Cooper
authors.forename: A L
authors.initials: AL
authors.lastname: Schwartz
authors.forename: G A
authors.initials: GA
authors.lastname: Patterson
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology; D005240:Feasibility Studies; D015870:Gene Expression; D018014:Gene Transfer Techniques; D016040:Lung Transplantation / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011916:Rats, Inbred F344; D009927:Tissue and Organ Procurement; D014184:Transplantation, Homologous; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D001616:beta-Galactosidase