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Gene transfer and expression of a non-viral polycation-based vector in CD4+ cells

Gene Therapy volume 6, pages 1774–1778 (1999)Cite this article

Abstract

CD4-selective targeting of an antibody-polycation-DNA complex was investigated. The complex was synthesized with the anti-CD4 monoclonal antibody B-F5, polylysine268 (pLL) and either the pGL3 control vector containing the luciferase reporter gene or the pGeneGrip vector containing the green fluorescent protein (GFP) gene. B-F5-pLL-DNA complexes inhibited the binding of 125I-B-F5 to CD4+Jurkat cells, while complexes synthesised either without B-F5 or using a non-specific mouse IgG1 antibody had little or no effect. Expression of the luciferase reporter gene was achieved in Jurkat cells using the B-F5-pLL-pGL3 complex and was enhanced in the presence of PMA. Negligible luciferase activity was detected with the non-specific antibody complex in Jurkat cells or with the B-F5-pLL-pGL3 complex in the CD4− K-562 cells. Using complexes synthesised with the pGeneGrip vector, the transfection efficiency in Jurkat and K-562 cells was examined using confocal microscopy. More than 95% of Jurkat cells expressed GFP and the level of this expression was markedly enhanced by PMA. Negligible GFP expression was seen in K-562 cells or when B-F5 was replaced by a non-specific antibody. Using flow cytometry, fluorescein-labelled complex showed specific targeting to CD4+ cells in a mixed cell population from human peripheral blood. These studies demonstrate the selective transfection of CD4+ T-lymphoid cells using a polycation-based gene delivery system. The complex may provide a means of delivering anti-HIV gene therapies to CD4+ cells in vivo.

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Acknowledgements

We wish to thank John Wijdenes of Diaclone, Besançon, France for his generous gift of the antibody, B-F5. This work was supported in part by the National Health and Medical Research Council of Australia, the Raine Medical Research Foundation and the AIDS Trust of Australia.

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Authors and Affiliations

  1. Department of Pharmacology, University of Western Australia, Western Australia, Australia

    R L Puls & R F Minchin

  2. Laboratory for Cancer Medicine, Royal Perth Hospital, Perth, Western Australia, Australia

    R F Minchin

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  1. R L Puls
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  2. R F Minchin
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Puls, R., Minchin, R. Gene transfer and expression of a non-viral polycation-based vector in CD4+ cells. Gene Ther 6, 1774–1778 (1999). https://doi.org/10.1038/sj.gt.3301022

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