title: Expression of an anti-CD4 single-chain antibody fragment from the donor cornea can prolong corneal allograft survival in inbred rats.
abstract: OBJECTIVE
  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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Ophthalmology, Flinders Medical Centre, Bedford Park, Adelaide, SA 5042, Australia.
authors.forename: Sarah Louise
authors.initials: SL
authors.lastname: Appleby
authors.forename: Claire F
authors.initials: CF
authors.lastname: Jessup
authors.forename: Lauren A
authors.initials: LA
authors.lastname: Mortimer
authors.forename: Kirsty
authors.initials: K
authors.lastname: Kirk
authors.forename: Helen M
authors.initials: HM
authors.lastname: Brereton
authors.forename: Douglas John
authors.initials: DJ
authors.lastname: Coster
authors.forename: Chuan K
authors.initials: CK
authors.lastname: Tan
authors.forename: Keryn A
authors.initials: KA
authors.lastname: Williams
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D003315:Cornea / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation / Q000502:physiology*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors; D006085:Graft Survival / Q000502:physiology*; D015948:Keratoplasty, Penetrating; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D011920:Rats, Inbred WF; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology; D014019:Tissue Donors; D014162:Transfection; D014184:Transplantation, Homologous
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D057127:Single-Chain Antibodies; C054562:yellow fluorescent protein, Bacteria