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.
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References
Steinman, R. M., Hawiger, D., and Nussenzweig, M. C. (2003) Tolerogenic dendritic cells. Annu. Rev. Immunol. 21, 767–811.
Morelli, A. E. and Thomson, A. W. (2003) Dendritic cells: regulators of alloimmunity and opportunities for tolerance induction. Immunol. Rev. 196, 125–146.
Fairchild, P. J. and Austyn, J. M. (1990) Thymic dendritic cells: phenotype and function. Int. Rev. Immunol. 6, 187–196.
Fairchild, P. J. and Waldmann, H. (2000) Dendritic cells and prospects for transplantation tolerance. Curr. Opin. Immunol. 12, 528–535.
Waldmann, H., Chen, T.-C., Graca, L., et al. (2006) Regulatory T cells in transplantation. Sem. Immunol. 18(2): 111–119.
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.
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.
Chen, L. (2004) Co-inhibitory molecules of the B7-CD28 family in the control of T-cell immunity. Nat. Rev. Immunol. 4, 336–347.
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.
Mellor, A. L. and Munn, D. H. (2004)IDO expression by dendritic cells: Tolerance and tryptophan catabolism. Nat. Rev. Immunol. 4, 762–774.
Jenne, L., Schuler, G., and Steinkasserer, A. (2001) Viral vectors for dendritic cell-based immunotherapy. Trends Immunol. 22, 102–102.
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.
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.
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.
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.
Thomson, J. A., Itskovitz-Eldor, J., Shapiro, S. S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.
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.
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© 2007 Humana Press Inc., Totowa, NJ
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Fairchild, P.J., Nolan, K.F., Waldmann, H. (2007). Genetic Modification of Dendritic Cells Through the Directed Differentiation of Embryonic Stem Cells. In: Fairchild, P.J. (eds) Immunological Tolerance. Methods in Molecular Biology™, vol 380. Humana Press. https://doi.org/10.1007/978-1-59745-395-0_4
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DOI: https://doi.org/10.1007/978-1-59745-395-0_4
Publisher Name: Humana Press
Print ISBN: 978-1-58829-652-8
Online ISBN: 978-1-59745-395-0
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