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    Transferrin receptor-mediated gene transfer to the corneal endothelium.

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    Authors
    Tan, Peng H
    King, William J
    Chen, Daxin
    Awad, Hana M
    Mackett, Mike
    Lechler, Robert I
    Larkin, D Frank
    George, Andrew J
    Affiliation
    Department of Immunology, Imperial College, Hammersmith Hospital, London, UK.
    Issue Date
    2001-02-27
    
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    Abstract
    BACKGROUND: The application of gene therapy to prevent allograft rejection requires the development of noninflammatory vectors. We have therefore investigated the use of a nonviral system, transferrin-mediated lipofection, to transfer genes into the cornea with the aim of preventing corneal graft rejection. METHODS: Rabbit and human corneas were cultured ex vivo and transfected with either lipofection alone or in conjunction with transferrin. The efficiency of transfection, localization, and kinetics of marker gene expression were determined. Strategies to increase gene expression, using chloroquine and EDTA, were investigated. In addition to a marker gene, a gene construct encoding viral interleukin 10 (vIL-10) was transfected and its functional effects were examined in vitro. RESULTS: Transferrin, liposome, and DNA were demonstrated to interact with each other, forming a complex. This complex was found to deliver genes selectively to the endothelium of corneas resulting in gene expression. Treatment of corneas with chloroquine and EDTA increased the transfection efficiency eight-fold and threefold, respectively. We also demonstrated that constructs encoding vIL-10 could be delivered to the endothelium. Secreted vIL-10 was shown to be functionally active by inhibition of a mixed lymphocyte reaction. CONCLUSIONS: Our data indicate that transferrin-mediated lipofection is a comparatively efficient nonviral method for delivering genes to the corneal endothelium. Its potential for use in preventing graft rejection is shown by the ability of this system to induce vIL-10 expression at secreted levels high enough to be functional.
    Citation
    Transferrin receptor-mediated gene transfer to the corneal endothelium. 2001, 71 (4):552-60 Transplantation
    Journal
    Transplantation
    URI
    http://hdl.handle.net/10541/85739
    PubMed ID
    11258435
    Type
    Article
    Language
    en
    ISSN
    0041-1337
    Collections
    All Paterson Institute for Cancer Research

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