Prolongation of sheep corneal allograft survival by ex vivo transfer of the gene encoding interleukin-10

Transplantation. 2001 May 15;71(9):1214-20. doi: 10.1097/00007890-200105150-00006.


Background: Modification of a donor cornea by gene therapy ex vivo has potential to modulate irreversible rejection, the major cause of corneal graft failure. Our aim was to transfer the gene encoding mammalian IL-10 to ovine donor corneas and to determine subsequent orthotopic corneal allograft survival in an outbred sheep model.

Methods: The replicative capacity of ovine corneal endothelium was determined by autoradiography after deliberate injury. A replication-defective adenovirus was used to deliver the lacZ reporter gene to ovine corneas and transfected corneas were organ-cultured in vitro to allow transfection efficiency, duration of reporter gene expression, and toxicity attributable to the vector to be determined. A cDNA encoding full-length ovine IL-10 was cloned into an adenoviral vector that was used to transfect donor corneas ex vivo before transplantation. Orthotopic penetrating corneal transplantation was performed in outbred sheep.

Results: Sheep corneal endothelium was found to be essentially amitotic. Transfection of > 70% corneal endothelial cells was achieved with the viral vector and expression was maintained for 28 days in vitro. IL-10 mRNA was detectable in transfected, organ-cultured corneas for 21 days in vitro. Donor corneas transfected with cDNA encoding IL-10 showed significantly prolonged survival after penetrating keratoplasty (median 55 days, range 19 > or =300 days) compared with control corneas (median 20.5 days, range 18-32 days, P=0.011).

Conclusion: Local gene therapy-mediated expression of the immunomodulatory cytokine IL-10 has the potential to reduce the incidence of corneal graft rejection and to prolong corneal allograft survival.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Corneal Transplantation / immunology*
  • Endothelium, Corneal / metabolism
  • Gene Transfer Techniques
  • Genes, Reporter
  • Graft Rejection / genetics
  • Graft Survival / physiology
  • Interleukin-10 / genetics*
  • RNA, Messenger / metabolism
  • Sheep
  • Transplantation, Homologous / immunology


  • RNA, Messenger
  • Interleukin-10