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.

  • Research Article
  • Published:

Characterization of HIV-1 vectors with gammaretrovirus envelope glycoproteins produced from stable packaging cells

Gene Therapy volume 11, pages 591–598 (2004)Cite this article

Abstract

We have recently described a novel, stable human immunodeficiency virus type 1 (HIV-1) vector packaging system, STAR. High-titre HIV-1 vectors bearing gammaretrovirus envelopes (Env) are continuously produced from STAR cells. Here we compare the properties of such vectors, with the amphotropic murine leukaemia virus (MLV-A) Env, a modified gibbon ape leukaemia virus (GALV) Env and two modified versions of the cat endogenous retrovirus RD114 Env, produced from STAR cells, to transiently produced HIV-1 vectors with vesicular stomatitis virus G protein (VSV-G). Our results indicate that gammaretrovirus pseudotypes from STAR cells are relatively stable at 37°C and are resistant to inactivation by freeze/thaw cycling or incubation with human sera. HIV-1(VSV-G) was, however, sensitive to freeze/thaw when harvested in serum-free media and was readily inactivated in human sera. Furthermore, the titre of ‘gamma-retrovirus’ pseudotypes, but not HIV-1(VSV-G), could be increased by the use of a combination of polybrene and spinoculation. All pseudotypes could be efficiently concentrated, but soluble gammaretrovirus Env could act as an inhibitor of infection.

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
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Naldini L et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263–267.

    Article  CAS  PubMed  Google Scholar 

  2. Sparacio S, Pfeiffer T, Schaal H, Bosch V . Generation of a flexible cell line with regulatable, high-level expression of HIV Gag/Pol particles capable of packaging HIV-derived vectors. Mol Ther 2001; 3: 602–612.

    Article  CAS  PubMed  Google Scholar 

  3. Kaul M, Yu H, Ron Y, Dougherty JP . Regulated lentiviral packaging cell line devoid of most viral cis-acting sequences. Virology 1998; 249: 167–174.

    Article  CAS  PubMed  Google Scholar 

  4. Klages N, Zufferey R, Trono D . A stable system for the high-titer production of multiply attenuated lentiviral vectors. Mol Ther 2000; 2: 170–176.

    Article  CAS  PubMed  Google Scholar 

  5. Kim VN, Mitrophanous K, Kingsman SM, Kingsman AJ . Minimal requirement for a lentivirus vector based on human immunodeficiency virus type 1. J Virol 1998; 72: 811–816.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Kotsopoulou E et al. A Rev-independent human immunodeficiency virus type 1 (HIV-1)-based vector that exploits a codon-optimized HIV-1 gag-pol gene. J Virol 2000; 74: 4839–4852.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Zufferey R et al. Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotechnol 1997; 15: 871–875.

    Article  CAS  PubMed  Google Scholar 

  8. Ikeda Y et al. Continuous high-titer HIV-1 vector production. Nat Biotechnol 2003; 21: 569–572.

    Article  CAS  PubMed  Google Scholar 

  9. Miller AD et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J Virol 1991; 65: 2220–2224.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Cosset FL et al. High-titer packaging cells producing recombinant retroviruses resistant to human serum. J Virol 1995; 69: 7430–7436.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Sommerfelt MA, Weiss RA . Receptor interference groups of 20 retroviruses plating on human cells. Virology 1990; 176: 58–69.

    Article  CAS  PubMed  Google Scholar 

  12. Uckert W et al. Efficient gene transfer into primary human CD8+ T lymphocytes by MuLV-10A1 retrovirus pseudotype. Hum Gene Ther 2000; 11: 1005–1014.

    Article  CAS  PubMed  Google Scholar 

  13. Kelly PF et al. Highly efficient gene transfer into cord blood nonobese diabetic/severe combined immunodeficiency repopulating cells by oncoretroviral vector particles pseudotyped with the feline endogenous retrovirus (RD114) envelope protein. Blood 2000; 96: 1206–1214.

    CAS  PubMed  Google Scholar 

  14. Hanawa H et al. Comparison of various envelope proteins for their ability to pseudotype lentiviral vectors and transduce primitive hematopoietic cells from human blood. Mol Ther 2002; 5: 242–251.

    Article  CAS  PubMed  Google Scholar 

  15. Blaese RM et al. T-lymphocyte directed gene therapy for ADA-SCID: initial trail results after 4 years. Science 1995; 270: 475–480.

    Article  CAS  PubMed  Google Scholar 

  16. Bordingnon C et al. Gene therapy in peripheral blood lymphocytes and bone marrow for ADA-immunodeficient patients. Science 1995; 270: 470–475.

    Article  Google Scholar 

  17. Cavazzana-Calvo M et al. Gene therapy of human severe combined immunodeficiency (SCID)-X1 disease. Science 2000; 288: 669–672.

    Article  CAS  PubMed  Google Scholar 

  18. Abonour R et al. Efficient retrovirus-mediated transfer of the multidrug resistance 1 gene into autologous human long-term repopulating hematopoietic stem cells. Nat Med 2000; 6: 652–658.

    Article  CAS  PubMed  Google Scholar 

  19. Pinter A et al. Localization of the labile disulfide bond between SU and TM of the murine leukemia virus envelope protein complex to a highly conserved CWLC motif in SU that resembles the active-site sequence of thiol-disulfide exchange enzymes. J Virol 1997; 71: 8073–8077.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Gliniak BC, Kozak SL, Jones RT, Kabat D . Disulfide bonding controls the processing of retroviral envelope glycoproteins. J Biol Chem 1991; 266: 22991–22997.

    CAS  PubMed  Google Scholar 

  21. Takeuchi Y et al. Type C retrovirus inactivation by human complement is determined by both the viral genome and the producer cell. J Virol 1994; 68: 8001–8007.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Christodoulopoulos I, Cannon PM . Sequences in the cytoplasmic tail of the gibbon ape leukemia virus envelope protein that prevent its incorporation into lentivirus vectors. J Virol 2001; 75: 4129–4138.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Stitz J et al. Lentiviral vectors pseudotyped with envelope glycoproteins derived from gibbon ape leukemia virus and murine leukemia virus 10A1. Virology 2000; 273: 16–20.

    Article  CAS  PubMed  Google Scholar 

  24. Sandrin V et al. Lentiviral vectors pseudotyped with a modified RD114 envelope glycoprotein show increased stability in sera and augmented transduction of primary lymphocytes and CD34+ cells derived from human and nonhuman primates. Blood 2002; 100: 823–832.

    Article  CAS  PubMed  Google Scholar 

  25. Marandin A et al. Retrovirus-mediated gene transfer into human CD34+38low primitive cells capable of reconstituting long-term cultures in vitro and nonobese diabetic-severe combined immunodeficiency mice in vivo. Hum Gene Ther 1998; 9: 1497–1511.

    Article  CAS  PubMed  Google Scholar 

  26. Duisit G, Salvetti A, Moullier P, Cosset FL . Functional characterization of adenoviral/retroviral chimeric vectors and their use for efficient screening of retroviral producer cell lines. Hum Gene Ther 1999; 10: 189–200.

    Article  CAS  PubMed  Google Scholar 

  27. Takeuchi Y et al. Sensitization of cells and retroviruses to human serum by (alpha 1-3) galactosyltransferase. Nature 1996; 379: 85–88.

    Article  CAS  PubMed  Google Scholar 

  28. DePolo NJ et al. VSV-G pseudotyped lentiviral vector particles produced in human cells are inactivated by human serum. Mol Ther 2000; 2: 218–222.

    Article  CAS  PubMed  Google Scholar 

  29. Takeuchi Y et al. Sensitization of rhabdo-, lenti-, and spumaviruses to human serum by galactosyl(alpha1-3)galactosylation. J Virol 1997; 71: 6174–6178.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Welsh RM, O'Donnell CL, Reed DJ, Rother RP . Evaluation of the Galalpha1-3Gal epitope as a host modification factor eliciting natural humoral immunity to enveloped viruses. J Virol 1998; 72: 4650–4656.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Reiser J . Production and concentration of pseudotyped HIV-1-based gene transfer vectors. Gene Therapy 2000; 7: 910–913.

    Article  CAS  PubMed  Google Scholar 

  32. VandenDriessche T, Naldini L, Collen D, Chuah MK . Oncoretroviral and lentiviral vector-mediated gene therapy. Methods Enzymol 2002; 346: 573–589.

    Article  CAS  PubMed  Google Scholar 

  33. Cosset FL et al. Retroviral retargeting by envelopes expressing an N-terminal binding domain. J Virol 1995; 69: 6314–6322.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Slingsby JH et al. Analysis of 4070A envelope levels in retroviral preparations and effect on target cell transduction efficiency. Hum Gene Ther 2000; 11: 1439–1451.

    Article  CAS  PubMed  Google Scholar 

  35. 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 

  36. Pinter A et al. Structural domains of endogenous murine leukemia virus gp70s containing specific antigenic determinants defined by monoclonal antibodies. Virology 1982; 116: 499–516.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by Biotechnology and Biological Sciences Research Council, Oxford BioMedica (UK) Ltd, Cancer Research UK and the Medical Research Council, UK. We thank Nigel Field for assistance in the column fractionation experiments.

Author information

Authors and Affiliations

  1. Department of Immunology and Molecular Pathology, Windeyer Institute of Medical Sciences, University College London, London, UK

    BL Strang, Y Ikeda, MKL Collins & Y Takeuchi

  2. Vectorologie Retrovirale et Therapie Genique, Ecole Normale Superieure de Lyon, Lyon, France

    F-L Cosset

Authors
  1. BL Strang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y Ikeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. F-L Cosset
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. MKL Collins
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Y Takeuchi
    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

Strang, B., Ikeda, Y., Cosset, FL. et al. Characterization of HIV-1 vectors with gammaretrovirus envelope glycoproteins produced from stable packaging cells. Gene Ther 11, 591–598 (2004). https://doi.org/10.1038/sj.gt.3302189

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302189

Keywords

Search

Advanced search

Quick links