Genetic Engineering of Dendritic Cells Using Retrovirus-Based Gene Transfer Techniques

  • Seema S. Ahuja
Part of the Methods in Molecular Biology book series (MIMB, volume 156)


Dendritic cells (DC) can be genetically engineered to constitutively express a gene of interest that could be either an immune-modulating cytokine, or an antigen (Ag) derived from a tumor/pathogen. There are numerous strategies for ex vivo transfer of genes or Ags into DCs. These include nonviral techniques (such as electroporation and liposomes) or recombinant viral-based vectors (e.g., retrovirus, adenovirus, herpes simplex virus, and avian and vaccinia viruses) (1-4 ). The major advantages of retrovirus vectors are that they are effective in achieving stable and highly efficient transduction of genes into primary cells, such as DCs/monocytes/macrophages (see Note 1). The major disadvantages are that the cells to be transduced must be in mitosis (i.e., actively dividing for reverse transcription and stable viral integration), and small size genes are more efficiently transduced than larger ones.


Antibiotic Resistance Gene Multiple Cloning Site Transduction Efficiency Polymerase Chain Reaction Amplicon Viral Supernatant 
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  1. 1.
    Danos, O. and Mulligan, R. C. (1988) Safe and efficient generation of recombi-nant retroviruses with amphotropic and ecotropic host ranges. Proc. Natl. Acad. Sci. USA 85, 6460–6464.CrossRefPubMedGoogle Scholar
  2. 2.
    Miller, A.D. and Rosman, G. J. (1989) Improved retroviral vectors for gene transfer and expression. Biotechniques 7, 980–990.PubMedGoogle Scholar
  3. 3.
    Bodine, D. (1997) Principles of gene therapy, in Cell Therapy (Ed Morstyn, G. and Sheridan, W.) Cambridge University Press, UK, pp. 91–109.Google Scholar
  4. 4.
    Cepko, C., Altshuler, D., Fekete, D., and Wu, D. (1992) Transduction of genes using retrovirus vectors, in Short Protocols in Molecular Biology (Ausubel, F.M., ed.), John Wiley, New York, 9–30–9–45.Google Scholar
  5. 5.
    Ahuja, S. S., Mummidi, S., Malech, H. L., and Ahuja, S. K. (1998) Human dendritic cell (DC)-based anti-infective therapy: engineering DCs to secrete functional interferon-γ and interleukin-12. J. Immunol. 161, 868–876.PubMedGoogle Scholar
  6. 6.
    Ahuja, S. S., Reddick, R. L., Sato, N., Montalbo, E., Kostecki, V., Zhao, W., Dolan, M. J., Melby, P.C., and Ahuja, S. K. (1999) Dendritic cells (DC)-based anti-infective strategies: DCs engineered to secret IL-12 are a potent vaccine in a murine model of an intracellular infection. J. Immunol. 163, 3890–3897.PubMedGoogle Scholar
  7. 7.
    Ahuja, S. S., Brown, M. R., Fleischer, T. A., Ahuja, S. K., and Malech, H. L. (1996) Autocrine activation of hemopoietic progenitor-derived myelomonocytic cells by IFN-γ gene transfer. J. Immunol. 156, 4345–4353.PubMedGoogle Scholar
  8. 8.
    Specht, J. M., Wang, G., Do, M. T., Lam, J. S., Royal, R-E., Reeves, M. E., Rothenberg, S. A., and Hwu, P. (1997) Dendritic cells retrovirally trasduced with a model antigen gene are therapeutically effective against established pulmonary metastasis. J. Exp. Med. 186, 1213–1221.CrossRefPubMedGoogle Scholar
  9. 9.
    Nishioka, Y., Lu, L., Lotze, M. T., Thara, H., and Thompson, A. W. (1997) Effective tumor immunotherapy using bone marrow-derived dendritic cells genetically engineered to exprfess interleukin 12. J. Immunother. 20, 419–425.CrossRefGoogle Scholar
  10. 10.
    Bahnson, A. B., Dunigan, J. T., Baysal, B. E., Mohney, T., Atchison, R. W., Nimgaonkar, M. T., Ball, E. D., and Barranger, J. A.. (1995) Centrifugal enhancement of retroviral mediated gene transfer. J. Virol. Meth. 54, 131–143.CrossRefGoogle Scholar
  11. 11.
    Takayama, T., Nishioka, Y., Lu, L., Lotze, T., Tahara, H., and Thompson, A. W. (1998) Retroviral delivery of viral interleukin-10 into myeloid dendritic cells markedly inhibits their allostimulatory activity and promotes the induction of T cell hyporesponsiveness. Transplantation 66, 1567–1574.CrossRefPubMedGoogle Scholar
  12. 12.
    Lee, W-C., Zhang, C., Qian, S., Wan, Y., Gauldie, J., Mi, Z., Robbins, P. D., Thompson, A. W., and Lu, L. (1998) Phenotype, function and in-vivo migration and survival of allogeneic dendritic cell progenitors genetically engineered to express TGF-β1. Transplantation 66, 1810–1817.CrossRefPubMedGoogle Scholar
  13. 13.
    Davis, I. D.and Lotze, M. T. (1999) Cytokine gene therapy, in Dendritic Cells (Lotze, M. T. and Thompson, A. W., eds.), Academic Press, London, pp. 823–853.Google Scholar
  14. 14.
    Lieschke, G. J., Rao, P. K., Gately, M. K., and Mulligan, R. C. (1997) Bioactive murine and human interleukin IL-12 fusion proteins which retain antitumor activity in vivo. Nat. Biotech. 15, 35–40.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Seema S. Ahuja
    • 1
  1. 1.Department of MedicineUniversity of Texas Health Science Center at San AntonioSan Antonio

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