Financial disclosure: This work was supported by the NHMRC, the Ophthalmic Research Institute of Australia and the Flinders Medical Centre Foundation.
Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea
Article first published online: 28 FEB 2010
© 2010 The Authors. Journal compilation © 2010 Royal Australian and New Zealand College of Ophthalmologists
Clinical & Experimental Ophthalmology
Volume 38, Issue 4, pages 405–413, May/June 2010
How to Cite
Parker, D. G., Coster, D. J., Brereton, H. M., Hart, P. H., Koldej, R., Anson, D. S. and Williams, K. A. (2010), Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea. Clinical & Experimental Ophthalmology, 38: 405–413. doi: 10.1111/j.1442-9071.2010.02261.x
- Issue published online: 11 JUN 2010
- Article first published online: 28 FEB 2010
- Received 29 September 2009; accepted 12 January 2010.
- adenoviral vector;
- corneal transplantation;
- gene therapy;
- lentiviral vector
Background: Gene transfer to a donor cornea ex vivo can modulate corneal graft failure in experimental animal models. We compared a lentiviral vector (LV) carrying the transgene ovine interleukin 10 (IL10) with a comparable adenoviral vector (Ad) for its ability to transduce ovine and human corneas and to modulate ovine corneal allograft survival.
Methods: The LV carrying the ovine IL10 gene was used to transduce ovine and human corneas in vitro. LV-mediated gene expression in corneal endothelium was assessed by real-time quantitative reverse-transcriptase polymerase chain reaction, at varying doses and duration of transduction. The effect of ex vivo transduction of the donor cornea with LV-SV40-IL10 was assessed following orthotopic corneal transplantation in outbred sheep.
Results: Expression of IL10 mRNA in Ad-CMV-IL10-transduced ovine corneas was 103-fold higher than in LV-SV40-IL10-transduced corneas (P < 0.0001), and 107-fold higher than in non-transduced controls. IL10 was secreted rapidly from Ad-CMV-IL10-transduced, organ-cultured corneas, peaking at 13–15 days. IL10 secreted from LV-SV40-IL10-transduced corneas increased 20-fold compared with controls, but had not reached a plateau at 15 days. Gene expression driven by LV-SV40-IL10 varied with vector dose and transduction time, but was less than with Ad-CMV-IL10 at both mRNA and protein levels. Gene expression driven by LV-SV40-IL10 was faster in the human cornea than the ovine cornea. Corneal allograft survival was prolonged by a median of 7 days in the LV-SV40-IL10-treated recipients, compared with the control group (P = 0.026).
Conclusion: Although lentiviral vectors show some promise for corneal gene therapy, they are less efficient than adenoviral vectors.