Abstract
Dendritic cells (DCs) are uniquely specialized antigen-presenting cells (APC) that play critical roles in both the stimulation and regulation of immune responses, including T-cell responses to transplanted organs. The inherent tolerogenicity of non-activated or “immature” DCs is well documented. Importantly, the infusion of DCs that are made resistant to activating inflammatory stimuli by “conditioning” through exposure to clinically approved immunosuppressants, such as corticosteroids, deoxyspergualin, and recently, rapamycin (RAPA), has produced encouraging outcomes in experimental models. Indeed, the infusion of RAPA-conditioned, host-derived DCs, pulsed with alloantigen, prolongs allograft survival. In particular, when the RAPA-conditioned DCs are delivered repeatedly or in combination with a short course of immunosuppression indefinite allograft survival is observed, typically associated with increased Foxp3+ T-regulatory cells (Treg). Herein, we detail the procedures to generate and characterize RAPA-conditioned murine DCs (RAPA-DCs) ex vivo and in vivo. RAPA-DCs represent a pharmacologically conditioned DC population that promotes allograft survival and enriches for antigen-specific T-regulatory cells (Treg). DCs conditioned with immunosuppressive agents, like RAPA, represent novel and clinically applicable vectors or “negative” cellular vaccines, which can be loaded with donor antigen, and potentially used to promote/maintain organ transplant tolerance.
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Banchereau, J., and Steinman, R.M. (1998) Dendritic cells and the control of immunity. Nature 392, 245–252.
Steinman, R.M., Hawiger, D., and Nussenzweig, M.C. (2003) Tolerogenic dendritic cells. Annu Rev Immunol 21, 685–711.
Banchereau, J., Briere, F., Caux, C., Davoust, J., Lebecque, S., Liu, Y.J., Pulendran, B., and Palucka, K. (2000) Immunobiology of dendritic cells. Annu Rev Immunol 18, 767–811.
Lotze, M.T., and Thomson, A.W., Dendritic Cells: Biology and Clinical Applications, pp. 1–794, second edition, M.T. Lotze and A.W. Thomson (Eds.), 2001, London, San Diego: Academic Press.
Morelli, A.E., and Thomson, A.W. (2003) Dendritic cells: regulators of alloimmunity and opportunities for tolerance induction. Immunol Rev 196, 125–146.
Morelli, A.E., and Thomson, A.W. (2007) Tolerogenic dendritic cells and the quest for transplant tolerance. Nat Rev Immunol 7, 610–621.
Fehervari, Z., and Sakaguchi, S. (2004) Control of Foxp3+CD25+CD4+ regulatory cell activation and function by dendritic cells. Int Immunol 16, 1769–1780.
Wing, K., Fehérvári, Z., and Sakaguchi, S. (2006) Emerging possibilities in the development and function of regulatory T cells. Int Immunol 18, 991–1000.
Levings, M.K., Gregori, S., Tresoldi, E., Cazzaniga, S., Bonini, C., and Roncarolo, M.G. (2005) Differentiation of Tr1 cells by immature dendritic cells requires IL-10 but not CD25+CD4+ Tr cells. Blood 105, 1162–1169.
Ueno, H., Klechevsky, E., Morita, R., Aspord, C., Cao, T., Matsui, T., Di Pucchio, T., Connolly, J., Fay, J.W., Pascual, V., Palucka, A.K., and Banchereau, J. (2007) Dendritic cell subsets in health and disease. Immunol Rev 219, 118–142.
Lotze, M.T., Zeh, H.J., Rubartelli, A., Sparvero, L.J., Amoscato, A.A., Washburn, N.R., Devera, M.E., Liang, X., Tör, M., and Billiar, T. (2007) The grateful dead: damage-associated molecular pattern molecules and reduction/oxidation regulate immunity. Immunol Rev 220, 60–81.
Fujii, S., Liu, K., Smith, C., Bonito, A.J., and Steinman, R.M. (2004) The linkage of innate to adaptive immunity via maturing dendritic cells in vivo requires CD40 ligation in addition to antigen presentation and CD80/86 costimulation. J Exp Med 199, 1607–1618.
Steinman, R.M., and Banchereau, J. (2007) Taking dendritic cells into medicine. Nature 449, 419–426.
Shortman, K., and Naik, S.H. (2007) Steady-state and inflammatory dendritic-cell development. Nat Rev Immunol 7, 19–30.
Sehgal, S.N. (1998) Rapamune (RAPA, rapamycin, sirolimus): mechanism of action immunosuppressive effect results from blockade of signal transduction and inhibition of cell cycle progression. Clin Biochem 31, 335–340.
Kahan, B.D., and Camardo, J.S. (2001) Rapamycin: clinical results and future opportunities. Transplantation 72, 1181–1193.
Hackstein, H., Taner, T., Zahorchak, A.F., Morelli, A.E., Logar, A.J., Gessner, A., and Thomson, A.W. (2003) Rapamycin inhibits IL-4-induced dendritic cell maturation in vitro and dendritic cell mobilization and function in vivo. Blood 101, 4457–4463.
Woltman, A.M., van der Kooij, S.W., Coffer, P.J., Offringa, R., Daha, M.R., and van Kooten, C. (2003) Rapamycin specifically interferes with GM-CSF signaling in human dendritic cells, leading to apoptosis via increased p27KIP1 expression. Blood 101, 1439–1445.
Monti, P., Mercalli, A., Leone, B.E., Valerio, D.C., Allavena, P., and Piemonti, L. (2003) Rapamycin impairs antigen uptake of human dendritic cells. Transplantation 75, 137–145.
Taner, T., Hackstein, H., Wang, Z., Morelli, A.E., and Thomson, A.W. (2005) Rapamycin-treated, alloantigen-pulsed host dendritic cells induce Ag-specific T cell regulation and prolong graft survival. Am J Transplant 5, 228–236.
Turnquist, H.R., Raimondi, G., Zahorchak, A.F., Fischer, R.T., Wang, Z., and Thomson, A.W. (2007) Rapamycin-conditioned dendritic cells are poor stimulators of allogeneic CD4+ T cells, but enrich for antigen-specific Foxp3+ T regulatory cells and promote organ transplant tolerance. J Immunol 178, 7018–7031.
Horibe, E.K., Sacks, J., Unadkat, J., Raimondi, G., Wang, Z., Ikeguchi, R., Marsteller, D., Ferreira, L.M., Thomson, A.W., Lee, W.P., and Feili-Hariri, M. (2008) Rapamycin-conditioned, alloantigen-pulsed dendritic cells promote indefinite survival of vascularized skin allografts in association with T regulatory cell expansion. Transpl Immunol 18, 307–318.
Ikeguchi, R., Sacks, J.M., Unadkat, J.V., Solari, M., Horibe, E.K., Thomson, A.W., Lee, A.W., and Feili-Hariri, M. (2008) Long-Term survival of limb allografts induced by pharmacologically conditioned, donor alloantigen-pulsed dendritic cells without maintenance immunosuppression. Transplantation 85, 237–246.
Fedoric, B. and Krishnan, R. (2008) Rapamycin downregulates the inhibitory receptors ILT2, ILT3, ILT4 on human dendritic cells and yet induces T cell hyporesponsiveness independent of FoxP3 induction. Immunol Lett 8. (available ahead of print online 22 July 2008)
Turnquist, H.R., Sumpter, T.L., Tsung, A., Zahorchak, A.F., Nakao, A., Nau, G.J., Liew, F.Y., Geller, D.A., and Thomson, A.W. (2008) IL-1beta-driven ST2L expression promotes maturation resistance in rapamycin-conditioned dendritic cells. J Immunol 181, 62–72.
Sordi, V., Bianchi, G., Buracchi, C., Mercalli, A., Marchesi, F., D’Amico, G., Yang, C.H., Luini, W., Vecchi, A., Mantovani, A., Allavena, P., and Piemonti, L. (2006) Differential effects of immunosuppressive drugs on chemokine receptor CCR7 in human monocyte-derived dendritic cells: selective upregulation by rapamycin. Transplantation 82, 826–834.
Acknowledgments
These protocols were developed with the support of National Institutes of Health (NIH) grants to A.W.T. (R01AI41011 and R01AI60994). H.R.T. was supported by non-concurrent fellowships from the American Society of Transplantation and the NIH (T32CA082084 and F32AI072940). R.F. is in receipt of a research training fellowship from the NIH (T32 DK71492).
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Turnquist, H.R., Fischer, R.T., Thomson, A.W. (2010). Pharmacological Modification of Dendritic Cells to Promote Their Tolerogenicity in Transplantation. In: Naik, S. (eds) Dendritic Cell Protocols. Methods in Molecular Biology, vol 595. Humana Press. https://doi.org/10.1007/978-1-60761-421-0_8
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DOI: https://doi.org/10.1007/978-1-60761-421-0_8
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