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Expression of an anti-CD4 single-chain antibody fragment from the donor cornea can prolong corneal allograft survival in inbred rats
  1. Sarah Louise Appleby1,
  2. Claire F Jessup1,
  3. Lauren A Mortimer1,
  4. Kirsty Kirk1,
  5. Helen M Brereton1,
  6. Douglas John Coster1,
  7. Chuan K Tan2,
  8. Keryn A Williams1
  1. 1Department of Ophthalmology, Flinders University, Adelaide, South Australia, Australia
  2. 2Department of Paediatrics, University of Adelaide, Adelaide, South Australia, Australia
  1. Correspondence to Dr Keryn A Williams, Department of Ophthalmology, Flinders Medical Centre, Bedford Park, Adelaide, SA 5042, Australia; keryn.williams{at}


Aim To investigate whether expression of an anti-CD4 antibody fragment (scFv) by a lentivector-transduced donor cornea can prolong rat corneal allograft survival.

Methods Inbred Fischer 344 rats received penetrating corneal allografts from Wistar-Furth donors after a 3 h transduction of the donor cornea with a lentivector carrying anti-CD4scFv cDNA (Lv-CD4scFv), a lentivector carrying the reporter gene-enhanced yellow fluorescence protein (LV-eYFP), or an adenoviral vector carrying anti-CD4 scFv cDNA (Ad-CD4scFv). Unmodified controls were also performed. Graft survival was assessed by corneal clarity, and rejection was confirmed histologically.

Results In organ-cultured corneas, expression of anti-CD4 scFv was detected at 2 days post-transduction with the adenoviral vector, compared with 5 days post-transduction with the lentivector, and was 10-fold higher than the former. More inflammation was observed in Ad-CD4scFv-modified allografts than in Lv-CD4scFv-modified grafts at 15 days postsurgery (p=0.01). The median time to rejection for unmodified, LV-eYFP and Ad-CD4scFv grafts was day 17, compared with day 22 for Lv-CD4scFv grafts (p≤0.018).

Conclusion Donor corneas transduced with a lentiviral vector carrying anti-CD4scFv cDNA showed a modest but significant prolongation in graft survival compared with unmodified, Lv-eYFP and Ad-CD4scFv grafts. However, rejection still occurred in all Lv-CD4scFv grafts, indicating that sensitisation may have been delayed but was not prevented.

  • Cornea
  • Treatment other
  • Experimental &#8211 animal models
  • Experimental &#8211 laboratory

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