Skip to main content
SpringerLink
Log in

Methods for Retrovirus-Mediated Gene Transfer into Primary T-Lymphocytes

  • Protocol
Gene Therapy Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 7))

Abstract

Recombinant retroviruses are efficient vectors for introducing genes into many mammalian cell types. They are useful in the context of clinical as well as experimental applications, owing to their ability to generate high-titer and helper-free viral stocks. Retroviral vectors are especially appropriate for the transduction of primary lymphocytes, because gene transfer is mediated by nonimmunogenic vectors and stable vector integration in the target genome is achieved. Stable integration in cells undergoing clonal expansion ensures that the foreign genetic material will be faithfully transmitted to the cells’ progeny.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Miller, D. G., Adam, M. A., and Miller, A. D. (1990) Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection Mol Cell. Biol 10, 4239–4292.

    CAS  Google Scholar 

  2. Roe, T. Y., Reynolds, T. C., Yu, G., and Brown, P. O. (1993) Integration of murine leukemia virus DNA depends on mitosis. EMBO J. 12, 2099–2108.

    Article  CAS  Google Scholar 

  3. Kim, J. W., Closs, E. I., Albritton, L. M., and Cunningham, J. M. (1991) Transport of cationic amino acids by the mouse ecotropic retrovirus receptor. Nature 352, 726–728.

    Article  Google Scholar 

  4. Wang, H., Kavanaugh, M P., North, R. A, and Kabat, D. (1991) Cell-surface receptor for ecotropic murine retroviruses is a basic amino-acid transporter. Nature 352, 729–731.

    Article  CAS  Google Scholar 

  5. Miller, D. G, Edwards, R. H., and Miller, A. D. (1994) Cloning of the cellular receptor for amphotropic murine retroviruses reveals homology to that for the gibbon ape leukemia virus. Proc. Natl Acad. Sci USA 91, 78–82.

    Article  CAS  Google Scholar 

  6. van Zeijl, M., Johann, S. V., Closs, E., Cunningham, J., Eddy, R., Shows, T. B., and O’Hara, B. (1994) A human amphotropic retrovirus receptor is a second member of the gibbon ape leukemia virus receptor family. Proc Natl Acad Sci USA 91, 1168–1172.

    Article  Google Scholar 

  7. Cooper, N. R., Jensen, F. C., Welsh, R. M, Jr., and Oldstone, M. B. A. (1976) Lysis of RNA tumor viruses by human serum: direct antibody-independent triggering of the classical complement pathway. J Exp Med. 144, 970–982.

    Article  CAS  Google Scholar 

  8. Vieillard, V., Lauret, E., Rousseau, V., and De Maeyer, E. (1994) Blocking of retroviral infection at a step prior to reverse transcription in cells transformed to constitutively express interferon /gb. Proc Natl Acad Sci USA 91, 2689–2693.

    Article  CAS  Google Scholar 

  9. Zinkernagel, R. M. (1993) Immunity to viruses, in Fundumental Immunology, 3rd ed., Raven, New York, pp 1211–1250.

    Google Scholar 

  10. Traunecker, A, Luke, W., and Karjalainen, K (1988) Soluble CD4 molecules neutralize human immunodeficiency virus type 1. Nature 331, 84–86.

    Article  CAS  Google Scholar 

  11. Fathman, C G, Fitch, F. W, Denis, K. A., and Witte, O. N (1989) Long term culture of immunocompetent cells, in Fundamental Immunology, 2nd ed., Raven, New York, pp. 803–815.

    Google Scholar 

  12. Yamada, G., Kitamura, Y., Sonoda, H., Harada, H., Taki, S., Mulligan, R. C., Osawa, H., Diamantstein, T., Yokoyama, S., and Taniguchi, T. (1987) Retroviral expression of the human IL-2 gene in a murine T cell line results in cell growth autonomy and tumorigenicity. EMBO J. 6, 2705–2709.

    Article  CAS  Google Scholar 

  13. Rosenberg, S. A., Aegersold, P., Cornetta, K., Kasid, A., Morgan, R., Moen, R., Karson, E. M., Lotze, M. T., Yang, J. C., Topalian, S. L., Merino, M. J., Culver, K., Miller, A. D., Blaese, R. M., and Anderson, W. F. (1990) Gene transfer into humans: immunotherapy of patients with advanced melanoma using tumor infiltrating lymphocytes modified by retroviral gene transduction N Engl J. Med 323, 570–578.

    Article  CAS  Google Scholar 

  14. Rivière, I., Brose, K., and Mulligan, R. C. (1995) Effects of retroviral design on expression of human adenosine deaminase in murine bone marrow transplant recipients engrafted with genetically modified cells. Proc Natl Acad Sci USA 92, 6733–6737.

    Article  Google Scholar 

  15. Strair, R. K., Towle, M., Heald, P. W., and Smith, B. R. (1990) Retroviral-mediated transfer and expression of exogenous genes in primary lymphoid cells: assaying for a viral transactivator activity in normal and malignant cells. Blood 76, 1201–1208.

    Article  CAS  Google Scholar 

  16. Ferrari, G., Rossini, S., Giavazzi, R, Maggioni, D, Nobili, N., Soldati, M., Ungers, G., Mavilio, F., Gilboa, E., and Bordignon, C. (1991) An in vivo model of somatic cell gene therapy for human severe combined immunodeficiency. Science 251, 1363–1366.

    Article  CAS  Google Scholar 

  17. Culver, K., Cornetta, K., Morgan, R., Morecki, S., Aebersold, P., Kasid, A, Lotze, M., Rosenberg, S. A., Anderson, W F., and Blaese, R M. (1991) Lymphocytes as cellular vehicles for gene therapy in mouse and man. Proc Natl Acad. Sci USA 88, 3155–3159.

    Article  CAS  Google Scholar 

  18. Kavanaugh, M. P., Miller, D. G., Zhang, W., Law, W., Kozak, S L., Kabat, D, and Miller, A. D. (1994) Cell-surface receptors for gibbon ape leukemia virus and amphotropic murine retrovirus are inducible sodium-dependent phosphate symporters. Proc. Natl Acad. Sci. USA 91, 7071–7075.

    Article  CAS  Google Scholar 

  19. Sadelain, M. and Mulligan, R. C (1992) Efficient retroviral-mediated gene transfer into murine primary lymphocytes Ninth International Immunology Congress, Budapest.

    Google Scholar 

  20. Mavilio, F., Ferrari, G., Rossini, S., Nobili, N., Bonini, C., Casorati, G., Traversari, C., and Bordignon, C (1994) Peripheral blood lymphocytes as target cells of retroviral vector-mediated gene transfer. Blood 83, 1988–1997.

    Article  CAS  Google Scholar 

  21. Cullen, B. R., Lomedico, P. T, and Ju, G (1984) Transcriptional interference in avian retroviruses: implications for the promoter insertion model of leukemogenesis. Nature 307, 241–245.

    Article  CAS  Google Scholar 

  22. Bowtell, D. D., Cory, S., Johnson, G. R., and Gonda, T. J. (1988) Comparison of expression in hematopoietic cells by retroviral vectors carrying two genes. J. Virol 62, 2464–2473.

    Article  CAS  Google Scholar 

  23. Danos, O. and Mulligan, R. C (1988) Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host range Proc Natl Acad Sci. USA 85, 6460–6464.

    Article  CAS  Google Scholar 

  24. Miller, A. D., Garcia, J. V., von Suhr, N., Lynch, C. M., Wilson, C., and Eiden, M. V. (1991) Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J Virol 65, 2220–2224.

    Article  CAS  Google Scholar 

  25. Hantzopoulos, P. A., Suri, C., Glass, D. J., Goldfarb, M. P., and Yancopoulos, G D. (1994) The low affinity NGF receptor, p75, can collaborate with each of the TRKs to potentiate functional responses to the neurotrophins. Neuron 13, 187–201.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, USA

    Michel Sadelain

Authors
  1. Michel Sadelain
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Pittsburgh, PA, USA

    Paul D. Robbins

Rights and permissions

Reprints and permissions

Copyright information

© 1997 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Sadelain, M. (1997). Methods for Retrovirus-Mediated Gene Transfer into Primary T-Lymphocytes. In: Robbins, P.D. (eds) Gene Therapy Protocols. Methods in Molecular Medicine, vol 7. Humana, Totowa, NJ. https://doi.org/10.1385/0-89603-484-4:241

Download citation

  • DOI: https://doi.org/10.1385/0-89603-484-4:241

  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-0-89603-484-6

  • Online ISBN: 978-1-59259-591-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation