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Simian immunodeficiency virus vector pseudotypes differ in transduction efficiency and target cell specificity in brain

Gene Therapy volume 14, pages 1330–1343 (2007)Cite this article

Abstract

Lentiviral vectors have proven to be promising tools for transduction of brain cells in vivo and in vitro. In this study, we have examined the central nervous system (CNS) transduction efficiencies and patterns of a self-inactivating simian immunodeficiency virus (SIVmac)-derived lentiviral vector pseudotyped with glycoproteins from the vesicular stomatitis virus (VSV-G), the amphotropic murine leukemia virus (MLV4070Aenv), the lymphocytic choriomeningitis virus (LCMV-GP), the Ross River virus (RRV-GP) and the rabies virus (RV-G). All glycoproteins were efficiently incorporated into SIV virions, allowing efficient transduction of neuronal cell lines as well as of primary dissociated mouse brain cell cultures. After injection of highly concentrated vector stocks into the striatum of adult mice, quantitative analyses revealed high transduction efficiency with VSV-G pseudotypes, while LCMV-GP and RV-G pseudotypes exhibited moderate transduction efficiencies. MLV4070Aenv and RRV-GP pseudotypes, however, showed only weak levels of transduction after stereotactic injection into the brain. Regarding cell tropism in vivo, VSV-G-pseudotyped SIV vectors transduced neuronal as well as glial cells, whereas all other pseudotypes preferentially transduced neuroglial cells. In addition, we analyzed the influence of the central polypurine tract (cPPT) in context of the VSV-G-pseudotyped SIV transfer vector for infection of brain cells. Deletion of the cPPT sequence from the transfer vector decreased the in vivo transduction efficiency by fourfold, and, more importantly, this modification changed the transduction pattern, since these vectors were no longer able to infect neuronal cells in vivo. Vector injection into the brain did elicit a humoral immune response in the injected hemisphere; however, no gross signs of inflammation could be detected. Analysis of the biodistribution of the vector revealed that, besides the injected brain region, no vector-specific sequences could be detected in any of the organs evaluated. These data indicate SIV vectors as efficient gene delivery vehicles for the treatment of neurodegenerative diseases.

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Acknowledgements

We are grateful to Dr Überla, Ruhr-University Bochum, Germany, for providing plasmids pViCGΔBH and pSgpΔ2; to Dr Cosset, INSERM U758, Lyon, France, for plasmid pAXF; to Dr Burns, University of California San Diego, USA, for providing plasmid pHCMV-G; to Dr von Laer, Georg-Speyer-Haus, Frankfurt, Germany, for plasmid pHCMV–LCMV-GP; to Dr Sanders, Purdue University, West Lafayette, USA, for plasmid pRRVpcDNA3.1Zeo+; and to Dr Conzelmann, Ludwig-Maximilians-University, München, Germany for providing the Rabies virus G cDNA of strain SAD B19. We also thank Helga Petznek for help with mouse work, Doris Rosenfellner for help with histological analyses, and Christopher Krewenka from the Institute of Medical Chemistry for preparation of the primary cell cultures. Special thanks are due to Gerrit Jandl for excellent technical assistance. We very much appreciate Dr Kirsti Witter, Institute of Histology and Embryology, and Dr Herbert Weissenböck, Institute of Pathology, University of Veterinary Medicine Vienna, for helpful discussions on stereological analyses and pathological advice, respectively. Quantification of infected cells was supported by project MSM0021620819 awarded by the Ministry of Education, Youth and Sports of the Czech Republic.

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  1. B Liehl

    Present address: 5Current address: German Institute of Human Nutrition, Nuthetal, Germany,

Authors and Affiliations

  1. Research Institute for Virology and Biomedicine, University of Veterinary Medicine, Vienna, Austria

    B Liehl, J Hlavaty, H Unger & W H Günzburg

  2. Institute of Medical Chemistry, University of Veterinary Medicine, Vienna, Austria

    R Moldzio

  3. Department of Histology and Embryology, Faculty of Medicine Pilsen, Charles University Prague, Pilsen, Czech Republic

    Z Tonar

  4. Austrianova Biotechnology GmbH, Vienna, Austria

    B Salmons & M Renner

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Liehl, B., Hlavaty, J., Moldzio, R. et al. Simian immunodeficiency virus vector pseudotypes differ in transduction efficiency and target cell specificity in brain. Gene Ther 14, 1330–1343 (2007). https://doi.org/10.1038/sj.gt.3302988

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