2.50
Hdl Handle:
http://hdl.handle.net/10541/85739
Title:
Transferrin receptor-mediated gene transfer to the corneal endothelium.
Authors:
Tan, Peng H; King, William J; Chen, Daxin; Awad, Hana M; Mackett, Mike; Lechler, Robert I; Larkin, D Frank; George, Andrew J
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.
Affiliation:
Department of Immunology, Imperial College, Hammersmith Hospital, London, UK.
Citation:
Transferrin receptor-mediated gene transfer to the corneal endothelium. 2001, 71 (4):552-60 Transplantation
Journal:
Transplantation
Issue Date:
27-Feb-2001
URI:
http://hdl.handle.net/10541/85739
PubMed ID:
11258435
Type:
Article
Language:
en
ISSN:
0041-1337
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorTan, Peng Hen
dc.contributor.authorKing, William Jen
dc.contributor.authorChen, Daxinen
dc.contributor.authorAwad, Hana Men
dc.contributor.authorMackett, Mikeen
dc.contributor.authorLechler, Robert Ien
dc.contributor.authorLarkin, D Franken
dc.contributor.authorGeorge, Andrew Jen
dc.date.accessioned2009-11-10T10:37:49Z-
dc.date.available2009-11-10T10:37:49Z-
dc.date.issued2001-02-27-
dc.identifier.citationTransferrin receptor-mediated gene transfer to the corneal endothelium. 2001, 71 (4):552-60 Transplantationen
dc.identifier.issn0041-1337-
dc.identifier.pmid11258435-
dc.identifier.urihttp://hdl.handle.net/10541/85739-
dc.description.abstractBACKGROUND: 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.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshChloroquine-
dc.subject.meshCytomegalovirus-
dc.subject.meshEndothelium, Corneal-
dc.subject.meshGene Expression-
dc.subject.meshGene Transfer Techniques-
dc.subject.meshGenetic Vectors-
dc.subject.meshHumans-
dc.subject.meshInterleukin-10-
dc.subject.meshPromoter Regions, Genetic-
dc.subject.meshRabbits-
dc.subject.meshReceptors, Transferrin-
dc.subject.meshTime Factors-
dc.subject.meshTransfection-
dc.subject.meshbeta-Galactosidase-
dc.titleTransferrin receptor-mediated gene transfer to the corneal endothelium.en
dc.typeArticleen
dc.contributor.departmentDepartment of Immunology, Imperial College, Hammersmith Hospital, London, UK.en
dc.identifier.journalTransplantationen

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