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Optimized lentiviral vector production and purification procedure prevents immune response after transduction of mouse brain

Gene Therapy volume 10, pages 1933–1940 (2003)Cite this article

Abstract

HIV-derived lentiviral vectors are efficient vehicula to deliver genes into the brain and hold great promise for future gene therapy of neurodegenerative disorders. However, administration of the current vector preparations in mouse brain was found to induce a systemic immune response to vector proteins and a modest inflammation in the brain. Moreover, serum antibodies from vector-treated animals were capable of partially neutralizing lentiviral vector-mediated transduction in cell culture. To avoid this unexpected immune reaction, we have optimized new vector production and purification protocols. Purification by sucrose gradient ultracentrifugation abolished the immune response, but vector titers also decreased substantially. Lentiviral vector production in the absence of serum in the cell culture medium equally reduced immunogenicity without affecting transduction efficiency. These results have important implications for future clinical use of lentiviral vectors, and for the use of lentiviral vectors to create animal models for neurodegenerative diseases that have an important neuroinflammatory component.

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Acknowledgements

We thank Sofie Willems for outstanding technical assistance with vector productions. The HIV-1-derived lentivirus vector system was a kind gift from Dr O Danos (Evry, France) and Dr D Trono (University of Geneva). The TRIP vectors were a generous gift from Dr P Charneau (Institut Pasteur, Paris). VB and ZD are postdoctoral fellows of the Flemish Fund for Scientific Research (FWO Vlaanderen). This work was funded by IDO Grant 98/006 and VIS Grant 00/010 from the KULeuven Research Council and the STWW Grant (980339) of the Flemish Institute supporting Scientific-Technological Research in Industry (IWT).

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Authors and Affiliations

  1. Laboratory for Experimental Neurosurgery and Neuroanatomy, Leuven, Belgium

    V Baekelandt, K Eggermont & B Nuttin

  2. Molecular Virology and Gene Therapy, Rega Institute for Medical Research, Leuven, Belgium

    M Michiels & Z Debyser

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  1. V Baekelandt
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  2. K Eggermont
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  3. M Michiels
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  4. B Nuttin
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  5. Z Debyser
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From the Gene Therapy Program, Katholieke Universiteit Leuven, Leuven, Belgium

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Baekelandt, V., Eggermont, K., Michiels, M. et al. Optimized lentiviral vector production and purification procedure prevents immune response after transduction of mouse brain. Gene Ther 10, 1933–1940 (2003). https://doi.org/10.1038/sj.gt.3302094

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