Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea

Douglas G. Parker, Douglas J. Coster, Helen M. Brereton, Prue H. Hart, Rachel Koldej, Donald S. Anson, Keryn Williams

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

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.

LanguageEnglish
Pages405-413
Number of pages9
JournalClinical and Experimental Ophthalmology
Volume38
Issue number4
DOIs
Publication statusPublished - 1 May 2010

Keywords

  • Adenoviral vector
  • Corneal transplantation
  • Gene therapy
  • Lentiviral vector

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Parker, Douglas G. ; Coster, Douglas J. ; Brereton, Helen M. ; Hart, Prue H. ; Koldej, Rachel ; Anson, Donald S. ; Williams, Keryn. / Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea. In: Clinical and Experimental Ophthalmology. 2010 ; Vol. 38, No. 4. pp. 405-413.
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abstract = "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.",
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Lentivirus-mediated gene transfer of interleukin 10 to the ovine and human cornea. / Parker, Douglas G.; Coster, Douglas J.; Brereton, Helen M.; Hart, Prue H.; Koldej, Rachel; Anson, Donald S.; Williams, Keryn.

In: Clinical and Experimental Ophthalmology, Vol. 38, No. 4, 01.05.2010, p. 405-413.

Research output: Contribution to journalArticle

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AU - Coster, Douglas J.

AU - Brereton, Helen M.

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AU - Anson, Donald S.

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AB - 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.

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