Genetic Modification of Dendritic Cells Through the Directed Differentiation of Embryonic Stem Cells

  • Paul J. Fairchild
  • Kathleen F. Nolan
  • Herman Waldmann
Part of the Methods in Molecular Biology™ book series (MIMB, volume 380)

Abstract

Recent years have witnessed a progressive acceptance of the dual role played by dendritic cells (DC) in the initiation of immune responses and their specific attenuation through the induction of immunological tolerance. Nevertheless, as terminally differentiated cells of the myeloid lineage, DC share with macrophages an inherent resistance to genetic modification, greatly restricting strategies available for studying their physiology and function. Consequently, little is known of the molecular interactions provided by DC that underlie the critical decision between tolerance and immunity. Embryonic stem (ES) cells are, by contrast, relatively amenable to genetic modification. Furthermore, their propensity for self-renewal, one of the cardinal features of a stem cell, permits cloning at the single cell level and the rational design of ES cell lines, uniformly expressing a desired, mutant phenotype. Here, we describe how another defining property of ES cells, their demonstrable pluripotency, may be harnessed for their directed differentiation along the DC pathway, enabling the generation of limitless numbers of DC faithfully expressing candidate genes of interest. The protocols we outline in this chapter may, therefore, offer new opportunities for dissecting the biology of DC and the molecular basis of their unique properties.

Key Words

Dendritic cell embryonic stem cell directed differentiation tolerance genetic modification 

References

  1. 1.
    Steinman, R. M., Hawiger, D., and Nussenzweig, M. C. (2003) Tolerogenic dendritic cells. Annu. Rev. Immunol. 21, 767–811.CrossRefGoogle Scholar
  2. 2.
    Morelli, A. E. and Thomson, A. W. (2003) Dendritic cells: regulators of alloimmunity and opportunities for tolerance induction. Immunol. Rev. 196, 125–146.PubMedCrossRefGoogle Scholar
  3. 3.
    Fairchild, P. J. and Austyn, J. M. (1990) Thymic dendritic cells: phenotype and function. Int. Rev. Immunol. 6, 187–196.PubMedCrossRefGoogle Scholar
  4. 4.
    Fairchild, P. J. and Waldmann, H. (2000) Dendritic cells and prospects for transplantation tolerance. Curr. Opin. Immunol. 12, 528–535.PubMedCrossRefGoogle Scholar
  5. 5.
    Waldmann, H., Chen, T.-C., Graca, L., et al. (2006) Regulatory T cells in transplantation. Sem. Immunol. 18(2): 111–119.CrossRefGoogle Scholar
  6. 6.
    Jonuleit, H., Schmitt, E., Schuler, G., Knop, J., and Enk, E. H. (2000) Induction of IL-10-producing, non-proliferating CD4+ T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells. J. Exp. Med. 192, 1213–1222.PubMedCrossRefGoogle Scholar
  7. 7.
    Mahnke, K., Qian, Y., Knop, J., and Enk, A. H. (2003) Induction of CD4+CD25+ regulatory T cells by targeting of antigens to immature dendritic cells. Blood 101, 4862–4869.PubMedCrossRefGoogle Scholar
  8. 8.
    Chen, L. (2004) Co-inhibitory molecules of the B7-CD28 family in the control of T-cell immunity. Nat. Rev. Immunol. 4, 336–347.PubMedCrossRefGoogle Scholar
  9. 9.
    Munn, D. H., Sharma, M. D., Lee, J. R., et al. (2002) Potential regulatory function of human dendritic cells expressing indoleamine 2,3-dioxygenase. Science 297, 1867–1870.PubMedCrossRefGoogle Scholar
  10. 10.
    Mellor, A. L. and Munn, D. H. (2004)IDO expression by dendritic cells: Tolerance and tryptophan catabolism. Nat. Rev. Immunol. 4, 762–774.PubMedCrossRefGoogle Scholar
  11. 11.
    Jenne, L., Schuler, G., and Steinkasserer, A. (2001) Viral vectors for dendritic cell-based immunotherapy. Trends Immunol. 22, 102–102.PubMedCrossRefGoogle Scholar
  12. 12.
    Morelli, A. E., Larregina, A. T., Ganster, et al. (2000) Recombinant adenovirus induces maturation of dendritic cells via an NF-κB-dependent pathway. J. Virol. 74, 9617–9628.PubMedCrossRefGoogle Scholar
  13. 13.
    Fairchild, P. J., Brook, F. A., Gardner, R. L., et al. (2000) Directed differentiation of dendritic cells from mouse embryonic stem cells. Curr. Biol. 10, 1515–1518.PubMedCrossRefGoogle Scholar
  14. 14.
    Fairchild, P. J., Nolan, K. F., and Waldmann, H. (2003) Probing dendritic cell function by guiding the differentiation of embryonic stem cells. Meth. Enzymol. 365, 169–186.PubMedCrossRefGoogle Scholar
  15. 15.
    Fairchild, P. J., Nolan, K. F., Graça, L., and Waldmann, H. (2003) Stable lines of genetically modified dendritic cells from embryonic stem cells. Transplantation 76, 606–608.PubMedCrossRefGoogle Scholar
  16. 16.
    Thomson, J. A., Itskovitz-Eldor, J., Shapiro, S. S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.PubMedCrossRefGoogle Scholar
  17. 17.
    Reubinoff, B. E., Pera, M. E, Fong, C. Y., Trounson, A., and Bongso, A. (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat. Biotechnol. 18, 399–404.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Paul J. Fairchild
    • 1
  • Kathleen F. Nolan
    • 1
  • Herman Waldmann
    • 1
  1. 1.Sir William Dunn School of PathologyUniversity of OxfordOxfordUK

Personalised recommendations