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.

  • Brief Communication
  • Published:

Adeno-retroviral chimeric viruses as in vivo transducing agents

Abstract

Several hybrid viral gene transfer systems have been described that exploit the favorable features of the two parent viral species. We have developed a hybrid adeno-retroviral vector system to generate a retroviral vector in situ. The system consists of adenoviruses encoding MoMLV gag.pol (Axtet.gag.pol), the VSV-G viral envelope (Axtet.VSV-G), the retroviral vector LXSN expressing the neomycin phosphotransferase gene (AV-LXSN) and a transcriptional regulator to control expression of gag.pol and envelope (AV-rtTA). In vitro, biologically active retroviral vector preparations were generated following adeno-retroviral transduction of 9L rat glioma cells. In vivo the transcomplementing adeno-retroviruses were co-administered intratumorally into subcutaneous 9L glioma tumors in rats and human A375 melanoma xenografts in nude mice. In the 9L rat model, G418R cell cultures were only obtained when 9L cells were harvested from tumors injected with all four transcomplementing adeno-retroviruses. Molecular analysis of DNA extracted from 9L G418R populations derived both in vitro and in vivo showed appropriate integration of the LXSN proviral sequence. Tumor cells were harvested 1, 3 and 4 weeks after adeno-retrovirus administration to the human A375 xenografts. The percentage of G418R colonies recovered from tumors transduced with all of the transcomplementing adeno-retroviruses increased with time, whereas no increase was observed in tumors transduced with AV-LXSN alone. DNA extracted from G418R A375 cell populations showed the presence of integrated proviral sequences only in animals that received the full complement of adeno-retroviruses. These results demonstrate that adenoviral vectors expressing transcomplementing genes for retroviral proteins and retroviral vector RNAs can be used for in situ transduction of target cells.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Fisher K et al. A novel adenovirus-adeno-associated virus hybrid vector that displays efficient rescue and delivery of the AAV genome Hum Gene Ther 1996 7: 2079–2087

    Article  CAS  PubMed  Google Scholar 

  2. Wang S, Vos J-M . A hybrid Herpesvirus infectious vector based on Epstein–Barr virus and Herpes Simplex virus type 1 for gene transfer into human cells in vitro and in vivo J Virol 1996 70: 8422–8430

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Li K, Garoff H . Production of infectious recombinant Moloney murine leukemia virus particles in BHK cells using Semliki Forest virus-derived RNA expression vectors Proc Natl Acad Sci USA 1996 93: 11658–11663

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Johnston KM et al. HSV/AAV hybrid amplicon vectors extend transgene expression in human glioma cells Hum Gene Ther 1997 8: 359–370

    Article  CAS  PubMed  Google Scholar 

  5. Fraefel C et al. Gene transfer into hepatocytes mediated by helper virus-free HSV/AAV hybrid vectors Mol Med 1997 3: 813–825

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Walfors J, Xanthopoulos KG, Morgan RA . Semliki forest virus-mediated production of retroviral vector RNA in retroviral packaging cells Human Gene Ther 1997 8: 2031–2041

    Article  Google Scholar 

  7. Feng M et al. Stable in vivo gene transduction via a novel adenoviral/retroviral chimeric vector Nat Biotech 1997 15: 866–870

    Article  CAS  Google Scholar 

  8. Ramsey WJ et al. Adenovirus vectors as transcomplementing templates for the production of replication defective retroviral vectors Biochem Biophys Res Commun 1998 246: 912–919

    Article  CAS  PubMed  Google Scholar 

  9. Giard DJ et al. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors J Natl Cancer Inst 1973 51: 1417–1423

    Article  CAS  PubMed  Google Scholar 

  10. Gershwin ME et al. Immunobiology of heterotransplanted human tumors in nude mice J Natl Cancer Inst 1977 58: 1455–1463

    Article  CAS  PubMed  Google Scholar 

  11. Hallenbeck PL et al. Replication of E1 deleted adenoviral gene therapy vectors in tumor cell lines (submitted for publication)

  12. Culver KW et al. In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors Science 1992 256: 1550–1552

    Article  CAS  PubMed  Google Scholar 

  13. Ram Z et al. Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells Nature Med 1997 3: 1354–1361

    Article  CAS  PubMed  Google Scholar 

  14. Yoshida Y, Hamada H . Adenovirus-mediated inducible gene expression through tetracycline-controllable transactivator with nuclear localization signal Biochem Biophys Res Commun 1997 230: 426–430

    Article  CAS  PubMed  Google Scholar 

  15. Yoshida Y, Emi N, Hamada H . VSV-G-pseudotyped retroviral packaging through adenovirus-mediated inducible gene expression Biochem Biophys Res Commun 1997 232: 379–382

    Article  CAS  PubMed  Google Scholar 

  16. Fishman GI, Kaplan M, Buttrick PM . Tetracycline-regulated cardiac gene expression in vivo J Clin Invest 1994 93: 1864–1868

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. McGrory WJ, Bautista DS, Graham FL . A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5 Virology 1988 163: 614–617

    Article  CAS  PubMed  Google Scholar 

  18. Graham FL, Prevec L . Adenovirus-based expression vectors and recombinant vaccines Biotechnology 1992 20: 363–390

    CAS  PubMed  Google Scholar 

  19. Okada T et al. Development of long lasting anti-tumor immunity followed by thymidine kinase-mediated killing of rat experimental glioma (submitted for publication)

  20. Barth RJJ et al. Unique murine tumor-associated antigens identified by tumor infiltrating lymphocytes J Immunol 1990 144: 1531–1537

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We wish to thank J Jagadeesh for technical assistance, Julie Lekstrom-Himes for providing pAV-rtTA and M Shah for access to unpublished data and R Morgan for advice, support and a critical appraisal of this manuscript. NJC is supported by a NIH, Fogarty Fellowship.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Caplen, N., Higginbotham, J., Scheel, J. et al. Adeno-retroviral chimeric viruses as in vivo transducing agents. Gene Ther 6, 454–459 (1999). https://doi.org/10.1038/sj.gt.3300835

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3300835

Keywords

This article is cited by

Search

Quick links