Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea (original) (raw)
- Paper
- Published: 01 July 1997
- DFP Larkin1,2,
- Z Fehervari1 nAff3,
- AP Byrnes3 nAff5,
- AM Rankin4,
- DO Haskard5,
- MJA Wood3,
- MJ Dallman4 &
- …
- AJT George1
Gene Therapy volume 4, pages 639–647 (1997) Cite this article
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Abstract
One attractive strategy to prevent or control allograft rejection is to genetically modify the donor tissue before transplantation. In this study, we have examined the feasibility of gene transfer to human corneal endothelium, using a number of recombinant adenovirus constructs. Ex vivo infection of human corneas with adenoviral vectors containing lacZ, under transcriptional control of either cytomegalovirus (CMV) or Rous sarcoma virus (RSV) promoters, provided high-level gene expression, which was largely restricted to endothelium. Expression of the reporter gene persisted at relatively high levels for up to 7 days, followed by a decline to indetectable levels by 28 days. RT-PCR analysis of lacZ transcription showed a similar picture with a short period (3–7 days) of RNA transcription after infection. In contrast, adenoviral DNA persisted for at least 56 days. Subsequently, we examined the expression of a potential therapeutic gene, CTLA-4 Ig fusion protein. Following infection of human corneas with adenoviral vectors encoding CTLA-4 Ig protein, high levels of the fusion protein were detected in corneal culture supernatants for up to 28 days. This protein was functionally active, as determined by binding to B7.1 (CD80)-expressing transfectants. This study suggests that genetic alteration of donor cornea before transplantation is a feasible approach for preventing or controlling allograft rejection. Similar gene-based strategies might also be feasible to prevent rejection of other transplanted tissues or organs.
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Author notes
- Z Fehervari
Present address: Department of Immunology, Glaxo Wellcome Medicines Research Centre, Gunnels Wood Road, Stevenage, SG1 2NY, UK - AP Byrnes
Present address: Department of Molecular Microbiology and Immunology, Johns Hopkins University, School of Public Health, 615 N Wolfe Street, Baltimore, 21205-2179, MD, USA
Authors and Affiliations
- Department of Immunology, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK
HB Oral, DFP Larkin, Z Fehervari & AJT George - Department of Pathology, Institute of Ophthalmology, London, UK
DFP Larkin - Department of Human Anatomy, University of Oxford, UK
AP Byrnes & MJA Wood - Department of Biology, Infection and Immunity Section, Imperial College of Science, Technology and Medicine, London, UK
AM Rankin & MJ Dallman - Department of Medicine, Cardiovascular Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK
DO Haskard
Authors
- HB Oral
- DFP Larkin
- Z Fehervari
- AP Byrnes
- AM Rankin
- DO Haskard
- MJA Wood
- MJ Dallman
- AJT George
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Oral, H., Larkin, D., Fehervari, Z. et al. Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea.Gene Ther 4, 639–647 (1997). https://doi.org/10.1038/sj.gt.3300443
- Received: 08 January 1997
- Accepted: 07 March 1997
- Issue date: 01 July 1997
- DOI: https://doi.org/10.1038/sj.gt.3300443