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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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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