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Evidence for nonspecific adsorption of targeted retrovirus vector particles to cells

Gene Therapy volume 8, pages 1088–1096 (2001)Cite this article

Abstract

The ability to specifically target a cell-type is important for the development of vectors for in vivo gene therapy. In order to produce retrovirus vectors targeting ovarian cancer cells, which specifically overexpress α folate receptor (αFR), a single chain antibody was fused as an N-terminal extension of the ecotropic and amphotropic murine leukemia virus (MLV) envelope glycoproteins. Vector particles bearing the modified glycoproteins were produced and analysed. Although conventional FACS studies indicated that viral particles bearing the modified Env could bind to ovarian cancer cells, targeted infection was not achieved. The initial step of virus–cell interaction was further studied using an immunofluorescence technique, which allows visualisation of single retrovirus particles. Vectors bearing chimeric or wild-type glycoproteins bound equally well to cells with or without the targeted receptor, although soluble chimeric glycoproteins bound specifically to FBP. Our results indicate that the incorporation of specific ligands to the virus envelope does not necessarily result in significant enhancement of vector particle binding. A similar interaction was also observed using Env-defective virus particles, suggesting that cellular factors incorporated into the lipid envelope play a dominant role in promoting initial adsorption of virus particles to cells. Significant implications arise from these observations on the interpretation of previous reports on ‘targeted’ vectors, and for the development of vectors for in vivo gene therapy protocols.

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Acknowledgements

This work was funded by GlaxoWellcome and the Medical Research Council. MP was supported by a EU TMR grant (contr number FMBICT961804). We thank Centocor for provision of Mov18 monoclonal antibody, Dr J Benard for Igrov1 cells, Dr LH Evans for 83A25 rat antibody, and Professor G Palu' for his continued support and interest.

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Author notes

  1. M Pizzato

    Present address: Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, 02115, MA, USA

Authors and Affiliations

  1. Wohl Virion Centre, Windeyer Institute of Medical Sciences, University College London, Stevenage, UK

    M Pizzato, RA Weiss & Y Takeuchi

  2. Virology Unit, GlaxoWellcome, Medicines Research Centre, Stevenage, UK

    M Pizzato & ED Blair

  3. Discovery Genetics, GlaxoWellcome Inc., NC, USA

    M Fling & J Kopf

  4. Department of Experimental Oncology, Unit of Molecular Therapies, Istituto Nazionale per lo Studio e la Cura dei Tumori, Milano, Italy

    A Tomassetti

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Pizzato, M., Blair, E., Fling, M. et al. Evidence for nonspecific adsorption of targeted retrovirus vector particles to cells. Gene Ther 8, 1088–1096 (2001). https://doi.org/10.1038/sj.gt.3301494

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