Antibody targeted gene transfer to endothelium

Journal of Gene Medicine - Tập 5 Số 4 - Trang 311-323 - 2003
Peng Tan1, Maria Manunta1, Navid Ardjomand1,2, S Xue1, Frank Larkin1,2, Dorian O. Haskard3, Kenneth M. Taylor4, Andrew J.T. George1
1Department of Immunology, Division of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 ONN, UK.
2Moorfields Eye Hospital, City Road, London EC1V 2PD, UK
3National Heart and Lung Institute, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 ONN, UK
4Department of Cardiothoracic Surgery, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 ONN, UK

Tóm tắt

AbstractBackgroundOne of the drawbacks of the currently available vectors for gene therapy is the lack of selectivity in gene delivery. We have therefore investigated a strategy to generate immunoliposomes to target non‐viral vectors to cell surface receptors on endothelium.Materials and methodsWe have developed a novel method of coupling antibodies (Abs) to liposomes complexed to DNA, using mild heat treatment to aggregate the immunoglobulin G (IgG). The interaction of plasmid DNA, liposomes and Abs was measured using a gel retardation assay and a resonant mirror biosensor. The size of the transfection complex was determined by light scattering, and the binding and internalization of the complex to cells was followed using flow cytometry. The transfection ability was tested on cell lines and primary cells in vitro and human corneal or vascular tissues ex vivo.ResultsThe interaction of antibodies with liposomes is relatively stable (t1/2 ≅ 45 min). The size of the liposome, Ab and DNA complex was found to be around 500 nm in 4% BSA. The addition of anti‐transferrin receptor Abs increased the internalization of the liposome‐DNA complex into cells. Abs against both transferrin receptor and E‐selectin were shown to augment transfection efficiency of liposomes to cell expressing the appropriate antigens. They are also shown to be efficient in mediating gene delivery to corneal and vascular tissues ex vivo.ConclusionsWe have shown that our novel vector is capable of in vitro and ex vivo gene delivery to cells and human tissues including cornea, artery and vein. In particular, an Ab against E‐selectin was effective at selectively delivering genes to activated endothelial cells expressing the adhesion molecule. Such a strategy will have applications for targeting these tissues prior to transplantation or autologous grafting, and, in the longer term, may allow in vivo targeting of gene therapy to inflammatory sites. Copyright © 2002 John Wiley & Sons, Ltd.

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Journal of Gene Medicine

Tập 5 Số 4

311-323

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