Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/80621
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Type: Journal article
Title: Expression of an anti-CD4 single-chain antibody fragment from the donor cornea can prolong corneal allograft survival in inbred rats
Author: Appleby, S.
Jessup, C.
Mortimer, L.
Kirk, K.
Brereton, H.
Coster, D.
Tan, C.
Williams, K.
Citation: British Journal of Ophthalmology, 2013; 97(1):101-105
Publisher: British Med Journal Publ Group
Issue Date: 2013
ISSN: 0007-1161
1468-2079
Statement of
Responsibility: 
Sarah Louise Appleby, Claire F Jessup, Lauren A Mortimer, Kirsty Kirk, Helen M Brereton, Douglas John Coster, Chuan K Tan, Keryn A Williams
Abstract: <h4>Aim</h4>To investigate whether expression of an anti-CD4 antibody fragment (scFv) by a lentivector-transduced donor cornea can prolong rat corneal allograft survival.<h4>Methods</h4>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.<h4>Results</h4>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).<h4>Conclusion</h4>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.
Keywords: Cornea
CD4-Positive T-Lymphocytes
Animals
Rats, Inbred F344
Rats, Inbred WF
Rats
Adenoviridae
Bacterial Proteins
Luminescent Proteins
Fluorescent Dyes
Enzyme-Linked Immunosorbent Assay
Keratoplasty, Penetrating
Transplantation, Homologous
Flow Cytometry
Transfection
Graft Survival
Gene Expression Regulation
Genes, Reporter
Genetic Vectors
Tissue Donors
Male
Single-Chain Antibodies
Rights: Copyright status unknown
DOI: 10.1136/bjophthalmol-2012-302360
Appears in Collections:Aurora harvest
Paediatrics publications

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