Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Paper
  • Published:

High efficiency in vitro gene transfer into vascular tissues using a pseudotyped retroviral vector without pseudotransduction

Gene Therapy volume 6, pages 1876–1883 (1999)Cite this article

Abstract

Murine leukemia virus (MuLV)-derived retroviral vectors have had limited application in vascular gene therapy because of low transduction efficiency of vascular tissues, both in vitro and in vivo. In this study, we compared the gene transfer efficiency of two retroviral vectors: amphotropic MuLV and a MuLV vector pseudotyped with the vesicular stomatitis virus G glycoprotein (VSV-G) envelope. Target vascular tissues included human endothelial cells (EC), smooth muscle cells (SMC) and saphenous veins (SV). Transduction efficiency of human EC and SMC was significantly higher for VSV-G pseudotyped MuLV vector (90%) than for Amphotropic MuLV (20%). Luminal surface en face analysis of transduced cultured SV showed a six- to 10-fold greater transduction efficiency with VSV-G pseudotyped MuLV. The tissue plasminogen activator (tPA) gene was transduced into EC using each vector. Four days following transduction, a 12-fold higher tPA antigen concentration and a 38-fold higher tPA enzymatic activity was measured from cells transduced with the VSV-G pseudotyped vectors as compared with the amphotropic MuLV. There was no detectable pseudotransduction (protein transfer) associated with the VSV-G MuLV vector. Both AZT inhibition of reverse transcriptase and cell division arrest by gamma irradiation inhibited transduction, indicating that viral transduction correlated with RNA reverse transcription and cell proliferation. MuLV pseudotyped with the VSV-G envelope glycoprotein is an effective retroviral vector for vascular gene therapy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Wilson JM et al. Implantation of vascular grafts lined with genetically modified endothelial cells Science 1989 244: 1344–1346

    Article  CAS  PubMed  Google Scholar 

  2. Nabel EG et al. Recombinant gene expression in vivo within endothelial cells of the arterial wall Science 1989 244: 1342–1344

    Article  CAS  PubMed  Google Scholar 

  3. Dichek DA, Lee SW, Nguyen NH . Characterization of recombinant plasminogen activator production by primate endothelial cells transduced with retroviral vectors Blood 1994 84: 504–516

    CAS  PubMed  Google Scholar 

  4. Geary RL et al. Gene transfer in baboons using prosthetic vascular grafts seeded with retrovirally transduced smooth muscle cells: a model for local and systemic gene therapy Hum Gene Ther 1994 5: 1211–1216

    Article  CAS  PubMed  Google Scholar 

  5. Ekhterae D, Stanley JC . Retroviral vector-mediated transfer and expression of human tissue plasminogen activator gene in human endothelial and vascular smooth muscle cells J Vasc Surg 1995 21: 953–962

    Article  CAS  PubMed  Google Scholar 

  6. Rade JJ, Schulick AH, Virmani R, Dichek DA . Local adenoviral-mediated expression of recombinant hirudin reduces neointima formation after arterial injury Nature Med 1996 2: 293–8

    Article  CAS  PubMed  Google Scholar 

  7. Laitinen M et al. Gene transfer into the carotid artery using an adventitial collar: comparison of the effectiveness of the plasmid-liposome complexes, retroviruses, pseudotyped retroviruses, and adenoviruses Hum Gene Ther 1997 8: 1645–1650

    Article  CAS  PubMed  Google Scholar 

  8. Rekhter MD et al. Gene transfer into normal and atherosclerotic human blood vessels (see comments) Circ Res 1998 82: 1243–1252

    Article  CAS  PubMed  Google Scholar 

  9. Miller AD, Miller DG, Garcia JV, Lynch CM . Use of retroviral vectors for gene transfer and expression Meth Enzymol 1993 217: 581–599

    Article  CAS  Google Scholar 

  10. Inaba M et al. Retroviral gene transfer: effects on endothelial cell phenotype J Surg Res 1998 78: 31–36

    Article  CAS  PubMed  Google Scholar 

  11. Zelenock JA et al. Improved retroviral transduction efficiency of vascular cells in vitro and in vivo during clinically relevant incubation periods using centrifugation to increase viral titers J Vasc Surg 1997 26: 119–127

    Article  CAS  PubMed  Google Scholar 

  12. Friedmann T, Yee JK . Pseudotyped retroviral vectors for studies of human gene therapy Nature Med 1995 1: 275–277

    Article  CAS  PubMed  Google Scholar 

  13. Schnell MJ et al. Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles Proc Natl Acad Sci USA 1996 93: 11359–11365

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Burns JC et al. Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells (see comments) Proc Natl Acad Sci USA 1993 90: 8033–8037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Chen ST et al. Generation of packaging cell lines for pseudotyped retroviral vectors of the G protein of vesicular stomatitis virus by using a modified tetracycline inducible system Proc Natl Acad Sci USA 1996 93: 10057–10062

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ory DS, Neugeboren BA, Mulligan RC . A stable human-derived packaging cell line for production of high titer retrovirus/vesicular stomatitis virus G pseudotypes Proc Natl Acad Sci USA 1996 93: 11400–11406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Arai T et al. A new system for stringent, high-titer vesicular stomatitis virus G protein-pseudotyped retrovirus vector induction by introduction of Cre recombinase into stable prepackaging cell lines J Virol 1998 72: 1115–1121

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Liu ML, Winther BL, Kay MA . Pseudotransduction of hepatocytes by using concentrated pseudotyped vesicular stomatitis virus G glycoprotein (VSV-G)-Moloney murine leukemia virus-derived retrovirus vectors: comparison of VSV-G and amphotropic vectors for hepatic gene transfer J Virol 1996 70: 2497–2502

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Gallardo HF, Tan C, Ory D, Sadelain M . Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes Blood 1997 90: 952–957

    CAS  PubMed  Google Scholar 

  20. An DS et al. High-efficiency transduction of human lymphoid progenitor cells and expression in differentiated T cells J Virol 1997 71: 1397–1404

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Sharma S, Cantwell M, Kipps TJ, Friedmann T . Efficient infection of a human T-cell line and of human primary peripheral blood leukocytes with a pseudotyped retrovirus vector Proc Natl Acad Sci USA 1996 93: 11842–11847

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Miller DG, Adam MA, Miller AD . Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection (published erratum appears in Mol Cell Biol 1992 Jan;12(1):433) Mol Cell Biol 1990 10: 4239–4242

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Sinclair AM et al. Interaction of vesicular stomatitis virus-G pseudotyped retrovirus with CD34+ and CD34+ CD38− hematopoietic progenitor cells Gene Therapy 1997 4: 918–927

    Article  CAS  PubMed  Google Scholar 

  24. Schlegel R, Tralka TS, Willingham MC, Pastan I . Inhibition of VSV binding and infectivity by phosphatidylserine: is phosphatidylserine a VSV-binding site? Cell 1983 32: 639–646

    Article  CAS  PubMed  Google Scholar 

  25. Conti C, Mastromarino P, Ciuffarella MG, Orsi N . Characterization of rat brain cellular membrane components acting as receptors for vesicular stomatitis virus. Brief report Arch Virol 1988 99: 261–269

    Article  CAS  PubMed  Google Scholar 

  26. Kavanaugh MP et al. Cell-surface receptors for gibbon ape leukemia virus and amphotropic murine retrovirus are inducible sodium-dependent phosphate symporters Proc Natl Acad Sci USA 1994 91: 7071–7075

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. DuBridge RB et al. Analysis of mutation in human cells by using an Epstein–Barr virus shuttle system Mol Cell Biol 1987 7: 379–387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Eton D et al. Genetic engineering of stent-grafts with a highly efficient pseudotyped retroviral vector J Vasc Surg 1999 29: 863–873

    Article  CAS  PubMed  Google Scholar 

  29. Voyta JC, Via DP, Butterfield CE, Zetter BR . Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein J Cell Biol 1984 99: 2034–2040

    Article  CAS  PubMed  Google Scholar 

  30. Han JY et al. Identification of envelope protein residues required for the expended host range of 10A1 murine leukemia virus J Virol 1997 71: 8103–8108

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Soneoka Y et al. A transient three-plasmid expression system for the production of high titer retroviral vectors Nucleic Acids Res 1995 23: 628–633

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. MacKrell AJ, Soong NW, Curtis CM, Anderson WF . Identification of a subdomain in the Moloney murine leukemia virus envelope protein involved in receptor binding J Virol 1996 70: 1768–1774

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Yee JK, Friedmann T, Burns JC . Generation of high-titer pseudotyped retroviral vectors with very broad host range Meth Cell Biol 1994 43 Pt A: 99–112

    Article  CAS  Google Scholar 

  34. Morgan RA et al. Analysis of the functional and host range-determining regions of the murine ectropic and amphotropic retrovirus envelope proteins J Virol 1993 67: 4712–4721

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by American Heart Association, Pacific Vascular Research Foundation, May R Wright Foundation, Culpepper Foundation, James H Zumberge Fund. We thank Dr Aileen Takahashi for the critical reading of the manuscript.

Author information

Authors and Affiliations

  1. Department of Surgery/Vascular Division, University of Southern California School of Medicine, Los Angeles, CA, USA

    H Yu, D Eton, Y Wang, S R Kumar, L Tang & T T Terramani

  2. Gene Therapy Laboratories, University of Southern California School of Medicine, Los Angeles, CA, USA

    C Benedict & W F Anderson

  3. House Ear Institute, Los Angeles, CA, USA

    G Hung

Authors
  1. H Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D Eton
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S R Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. L Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T T Terramani
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. C Benedict
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. G Hung
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. W F Anderson
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yu, H., Eton, D., Wang, Y. et al. High efficiency in vitro gene transfer into vascular tissues using a pseudotyped retroviral vector without pseudotransduction. Gene Ther 6, 1876–1883 (1999). https://doi.org/10.1038/sj.gt.3301019

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301019

Keywords

This article is cited by

Search

Advanced search

Quick links