Adenovirus mediated gene transfer into rat lung grafts at the time of harvest

Ralph A. Schmid, Masaaki Narita, Carlos H. Boasquevisque, Koei Ando, Mitchell D. Botney, Joel D. Cooper, Alan L. Schwartz, G. Alexander Patterson

Research output: Contribution to journalArticle

5 Scopus citations

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°C). 4 x 1011 viral particles of adenovirus 5 containing the E. coli lacZ reporter gene coding for β- galactosidase (AdCMV-β-Gal) were added to the last milliliter of the flash solution. Lung grafts were stored for 3.5 h at room temperature followed by syngeneic 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 (β-Gal substrate) and kept in x-Gal for 3 h at 37°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 β-Gal activity. Conclusion: 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.

Original languageEnglish
Pages (from-to)1023-1029
Number of pages7
JournalEuropean Journal of Cardio-thoracic Surgery
Volume11
Issue number6
DOIs
StatePublished - Jun 1 1997

Keywords

  • Adenovirus
  • Gene transfer
  • Lung transplantation
  • Rat

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