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Shuttle of lentiviral vectors via transplanted cells in vivo

Gene Therapy volume 12, pages 67–74 (2005)Cite this article

Abstract

Lentiviral vectors have turned out to be an efficient method for stable gene transfer in vitro and in vivo. Not only do fields of application include cell marking and tracing following transplantation in vivo, but also the stable delivery of biological active proteins for gene therapy. A variety of cells, however, need immediate transplantation after preparation, for example, to prevent cell death, differentiation or de-differentiation. Although these cells are usually washed several times following lentiviral transduction, there may be the risk of viral vector shuttle via transplanted cells resulting in undesired in vivo transduction of recipient cells. We investigated whether infectious lentiviral particles are transmitted via ex vivo lentivirally transduced cells. To this end, we explored potential viral shuttle via ex vivo lentivirally transduced cardiomyocytes in vitro and following transplantation into the brain and peripheral muscle. We demonstrate that, even after extensive washing, infectious viral vector particles can be detected in cell suspensions. Those lentiviral vector particles were able to transduce target cells in transwell experiments. Moreover, transmitted vector particles stably transduced resident cells of the recipient central nervous system and muscle in vivo. Our results of lentiviral vector shuttle via transduced cardiomyocytes are significant for both ex vivo gene therapy and for lentiviral cell tracing, in particular for investigation of stem cell differentiation in transplantation models and co-cultivation systems.

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References

  1. Naldini L et al. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci USA 1996; 93: 11382–11388.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Zennou V et al. Loss of viral fitness associated with multiple Gag and Gag-Pol processing defects in human immunodeficiency virus type 1 variants selected for resistance to protease inhibitors in vivo. J Virol 1998; 72: 3300–3306.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Fleury S et al. Multiply attenuated, self-inactivating lentiviral vectors efficiently deliver and express genes for extended periods of time in adult rat cardiomyocytes in vivo. Circulation 2003; 107: 2375–2382.

    Article  CAS  PubMed  Google Scholar 

  4. Stitz J et al. A novel lentivirus vector derived from apathogenic simian immunodeficiency virus. Virology 2001; 291: 191–197.

    Article  CAS  PubMed  Google Scholar 

  5. Blomer U et al. Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector. J Virol 1997; 71: 6641–6649.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Kafri T et al. Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors. Nat Genet 1997; 17: 314–317.

    Article  CAS  PubMed  Google Scholar 

  7. Scherr M, Eder M . Gene transfer into hematopoietic stem cells using lentiviral vectors. Curr Gene Ther 2002; 2: 45–55.

    Article  CAS  PubMed  Google Scholar 

  8. Englund U et al. The use of a recombinant lentiviral vector for ex vivo gene transfer into the rat CNS. Neuroreport 2000; 11: 3973–3977.

    Article  CAS  PubMed  Google Scholar 

  9. Condorelli G et al. Cardiomyocytes induce endothelial cells to trans-differentiate into cardiac muscle: implications for myocardium regeneration. Proc Natl Acad Sci USA 2001; 98: 10733–10738.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ericson C, Wictorin K, Lundberg C . Ex vivo and in vitro studies of transgene expression in rat astrocytes transduced with lentiviral vectors. Exp Neurol 2002; 173: 22–30.

    Article  CAS  PubMed  Google Scholar 

  11. Arsenijevic Y, Weiss S . Insulin-like growth factor-I is a differentiation factor for postmitotic CNS stem cell-derived neuronal precursors: distinct actions from those of brain-derived neurotrophic factor. J Neurosci 1998; 18: 2118–2128.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Arsenijevic Y et al. Isolation of multipotent neural precursors residing in the cortex of the adult human brain. Exp Neurol 2001; 170: 48–62.

    Article  CAS  PubMed  Google Scholar 

  13. Bonci D et al. ‘Advanced’ generation lentiviruses as efficient vectors for cardiomyocyte gene transduction in vitro and in vivo. Gene Therapy 2003; 10: 630–636.

    Article  CAS  PubMed  Google Scholar 

  14. Sakoda T, Kasahara N, Hamamori Y, Kedes L . A high-titer lentiviral production system mediates efficient transduction of differentiated cells including beating cardiac myocytes. J Mol Cell Cardiol 1999; 31: 2037–2047.

    Article  CAS  PubMed  Google Scholar 

  15. Zhao J et al. Lentiviral vectors for delivery of genes into neonatal and adult ventricular cardiac myocytes in vitro and in vivo. Basic Res Cardiol 2002; 97: 348–358.

    Article  CAS  PubMed  Google Scholar 

  16. Blömer U et al. Lentiviral gene transfer and stable transgene expression in cardiomyoctes in vitro and in vivo. Mol Ther 2002; 5: 242.

    Article  Google Scholar 

  17. Conti C, Mastromarino P, Orsi N . Role of membrane phospholipids and glycolipids in cell-to-cell fusion by VSV. Comp Immunol Microbiol Infect Dis 1991; 14: 303–313.

    Article  CAS  PubMed  Google Scholar 

  18. Carneiro FA et al. Membrane fusion induced by vesicular stomatitis virus depends on histidine protonation. J Biol Chem 2003; 278: 13789–13794.

    Article  CAS  PubMed  Google Scholar 

  19. Nagata S, Kurata T, Ueno Y, Chiba J . Vesicular stomatitis virus-mediated cell fusion subsequent to virus adsorption at different pH values. Jpn J Med Sci Biol 1991; 44: 171–180.

    Article  CAS  PubMed  Google Scholar 

  20. Takehara M . Effect of certain inhibitors of glycoprotein synthesis on cell fusion induced by vesicular stomatitis virus. Microbiol Immunol 1979; 23: 167–176.

    Article  CAS  PubMed  Google Scholar 

  21. Haas DL, Case SS, Crooks GM, Kohn DB . Critical factors influencing stable transduction of human CD34(+) cells with HIV-1-derived lentiviral vectors. Mol Ther 2000; 2: 71–80.

    Article  CAS  PubMed  Google Scholar 

  22. Zufferey R et al. Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 1998; 72: 9873–9880.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Zufferey R, Donello JE, Trono D, Hope TJ . Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. J Virol 1999; 73: 2886–2892.

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Scherr M et al. Quantitative determination of lentiviral vector particle numbers by real-time PCR. Biotechniques 2001; 31: 520 522, 524, passim.

    Article  CAS  PubMed  Google Scholar 

  25. Kofidis T et al. In vitro engineering of heart muscle: artificial myocardial tissue. J Thorac Cardiovasc Surg 2002; 124: 63–69.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the HILF program of the Hannover Medical School. Ulrike Blömer was supported by ‘Deutsche Stiftung Querschnittslähmung’ and Ina Gruh by the ‘Graduiertenförderung’ of the Hannover Medical School and Niedersachsen Stiftung/CORTISS. pHR'SIN-CEW, pCMVΔR8.91 and pMD.G were kind gifts of D Trono. We are grateful to M Schweizer, ZMNH Hamburg, for providing time on the confocal microscope and to Patricia Schult-Dietrich, who performed the p24 assay.

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  1. Neurosurgery–Spine Surgery, Osnabrück, Germany

    U Blömer

  2. Leibnitz Research Labs for Biotechnology and Artificial Organs (LEBAO), Department of Thoracic and Cardiovascular Surgery, Hannover Medical School, Germany

    U Blömer, I Gruh, H Witschel, A Haverich & U Martin

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Blömer, U., Gruh, I., Witschel, H. et al. Shuttle of lentiviral vectors via transplanted cells in vivo. Gene Ther 12, 67–74 (2005). https://doi.org/10.1038/sj.gt.3302384

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