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Enhanced pseudotyping efficiency of HIV-1 lentiviral vectors by a rabies/vesicular stomatitis virus chimeric envelope glycoprotein

Gene Therapy volume 19, pages 761–774 (2012)Cite this article

Abstract

Rabies virus glycoprotein (RVG) can pseudotype lentiviral vectors, although at a lower efficiency to that of vesicular stomatitis virus glycoprotein (VSVG). Transduction with VSVG-pseudotyped vectors of rodent central nervous system (CNS) leads to local neurotropic gene transfer, whereas with RVG-pseudotyped vectors additional disperse transduction of neurons located at distal efferent sites occurs via axonal retrograde transport. Attempts to produce high-titre RVG-pseudotyped lentiviral vectors for preclinical and clinical trials has to date been problematic. We have constructed several chimeric RVG/VSVG glycoproteins and found that a construct bearing the external/transmembrane domain of RVG and the cytoplasmic domain of VSVG shows increased incorporation onto HIV-1 lentiviral particles and has increased infectivity in vitro in 293T cells and in differentiated neuronal cell lines of human, rat and murine origin. Stereotactic application of vector pseudotyped with this RVG/VSVG chimera in the rat striatum resulted in efficient gene transfer at the site of injection showing both neuronal and glial tropism. Distal neuronal transduction in the substantia nigra, thalamus and olfactory bulb via retrograde axonal transport also occurs after intrastriatal administration of chimera-pseudotyped vectors at similar levels to that observed with a RVG-pseudotyped vector. This is the first report of distal transduction in the olfactory bulb. The enhanced pseudotyping with this envelope should enable easier production of higher-titre pseudotyped lentiviral vectors that exhibit efficient local and dispersed neuronal transduction in the CNS.

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Acknowledgements

We thank Dr Edward Wright (University College London, London, UK) for the kind gift of the SNB1 anti-rabies virus GP antibody, and Professor Richard Reynolds and Dr Owain Howell for use of and help with fluorescence microscopy to document histological staining of brain sections. We also thank Dr Egle Solito and Enrico Cristante for letting us use several antibodies. This work was funded by a Seventh Framework Programme European Research Council Advanced Grant No. 23314 to NDM supporting DCJC, AT and SME.

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  1. K Vevis

    Present address: 2Current address: UCL Institute of Ophthalmology, London, UK.,

  2. R I Asfahani

    Present address: 3Current address: Dubowitz Neuromuscular Centre, UCL Institute of Child Health, London, UK.,

Authors and Affiliations

  1. Department of Gene Therapy, Division of Experimental Medicine, Centre for Neurosciences, Faculty of Medicine, Imperial College London, Hammersmith Hospital, London, UK

    D C J Carpentier, K Vevis, A Trabalza, C Georgiadis, S M Ellison, R I Asfahani & N D Mazarakis

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  1. D C J Carpentier
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Correspondence to N D Mazarakis.

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Carpentier, D., Vevis, K., Trabalza, A. et al. Enhanced pseudotyping efficiency of HIV-1 lentiviral vectors by a rabies/vesicular stomatitis virus chimeric envelope glycoprotein. Gene Ther 19, 761–774 (2012). https://doi.org/10.1038/gt.2011.124

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