Abstract
Human monocyte-derived dendritic cells (moDC) are commonly used as a research tool to investigate interactions between antigen-presenting cells and T cells. Generation of these cells involves the isolation of CD14 positive monocytes from peripheral blood and their in vitro differentiation into immature moDC by the cytokines GM-CSF and IL-4. Their functional characteristics can then be manipulated by maturing these cells with a cocktail of agents, which can be tailored to induce either immune activating or tolerogenic properties. Here, we describe a protocol for the generation of moDC with stable tolerogenic function, referred to as tolerogenic dendritic cells. These cells have been developed as an immunotherapeutic tool for the treatment of autoimmune disease but have also proven useful to dissect mechanisms of T cell tolerance induction in vitro.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, Muramatsu S, Steinman RM (1992) Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med 176(6):1693–1702. https://doi.org/10.1084/jem.176.6.1693
Sallusto F, Lanzavecchia A (1994) Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha. J Exp Med 179(4):1109–1118. https://doi.org/10.1084/jem.179.4.1109
Romani N, Gruner S, Brang D, Kämpgen E, Lenz A, Trockenbacher B, Konwalinka G, Fritsch PO, Steinman RM, Schuler G (1994) Proliferating dendritic cell progenitors in human blood. J Exp Med 180(1):83–93. https://doi.org/10.1084/jem.180.1.83
Cella M, Engering A, Pinet V, Pieters J, Lanzavecchia A (1997) Inflammatory stimuli induce accumulation of MHC class II complexes on dendritic cells. Nature 388(6644):782–787. https://doi.org/10.1038/42030
Hilkens CM, Kalinski P, de Boer M, Kapsenberg ML (1997) Human dendritic cells require exogenous interleukin-12-inducing factors to direct the development of naive T-helper cells toward the Th1 phenotype. Blood 90(5):1920–1926. https://doi.org/10.1182/blood.V90.5.1920
de Jong EC, Vieira PL, Kalinski P, Kapsenberg ML (1999) Corticosteroids inhibit the production of inflammatory mediators in immature monocyte-derived DC and induce the development of tolerogenic DC3. J Leukoc Biol 66(2):201–204. https://doi.org/10.1002/jlb.66.2.201
Ten Brinke A, Hilkens CM, Cools N, Geissler EK, Hutchinson JA, Lombardi G, Lord P, Sawitzki B, Trzonkowski P, Van Ham SM, Martinez-Caceres EM (2015) Clinical use of Tolerogenic dendritic cells-harmonization approach in European collaborative effort. Mediat Inflamm 2015:471719. https://doi.org/10.1155/2015/471719
Hilkens CM, Isaacs JD, Thomson AW (2010) Development of dendritic cell-based immunotherapy for autoimmunity. Int Rev Immunol 29(2):156–183. https://doi.org/10.3109/08830180903281193
Navarro-Barriuso J, Mansilla MJ, Martínez-Cáceres EM (2018) Searching for the transcriptomic signature of immune tolerance induction-biomarkers of safety and functionality for Tolerogenic dendritic cells and regulatory macrophages. Front Immunol 9:2062. https://doi.org/10.3389/fimmu.2018.02062
Anderson AE, Swan DJ, Sayers BL, Harry RA, Patterson AM, von Delwig A, Robinson JH, Isaacs JD, Hilkens CM (2009) LPS activation is required for migratory activity and antigen presentation by tolerogenic dendritic cells. J Leukoc Biol 85(2):243–250. https://doi.org/10.1189/jlb.0608374
Anderson AE, Sayers BL, Haniffa MA, Swan DJ, Diboll J, Wang XN, Isaacs JD, Hilkens CM (2008) Differential regulation of naïve and memory CD4+ T cells by alternatively activated dendritic cells. J Leukoc Biol 84(1):124–133. https://doi.org/10.1189/jlb.1107744
Anderson AE, Swan DJ, Wong OY, Buck M, Eltherington O, Harry RA, Patterson AM, Pratt AG, Reynolds G, Doran JP, Kirby JA, Isaacs JD, Hilkens CM (2017) Tolerogenic dendritic cells generated with dexamethasone and vitamin D3 regulate rheumatoid arthritis CD4+ T cells partly via transforming growth factor-β1. Clin Exp Immunol 187(1):113–123. https://doi.org/10.1111/cei.12870
Spiering R, Jansen MAA, Wood MJ, Fath AA, Eltherington O, Anderson AE, Pratt AG, van Eden W, Isaacs JD, Broere F, Hilkens CM (2019) Targeting of tolerogenic dendritic cells to heat-shock proteins in inflammatory arthritis. J Transl Med 17:375. https://doi.org/10.1186/s12967-019-2128-4
Harry RA, Anderson AE, Isaacs JD, Hilkens CM (2010) Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis. Ann Rheum Dis 69(11):2042–2050. https://doi.org/10.1136/ard.2009.126383
Bell GM, Anderson AE, Diboll J, Reece R, Eltherington O, Harry RA, Fouweather T, MacDonald C, Chadwick T, McColl E, Dunn J, Dickinson AM, Hilkens CM, Isaacs JD (2017) Autologous tolerogenic dendritic cells for rheumatoid and inflammatory arthritis. Ann Rheum Dis 76(1):227–234. https://doi.org/10.1136/annrheumdis-2015-208456
Acknowledgements
The authors thank Ioana Nicoresu for demonstrating the layering of blood onto a density gradient medium (Fig. 1) and Daniel Beckers for critically reading the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Hilkens, C.M.U., Diboll, J., Cooke, F., Anderson, A.E. (2023). In Vitro Generation of Human Tolerogenic Monocyte-Derived Dendritic Cells. In: Baldari, C.T., Dustin, M.L. (eds) The Immune Synapse. Methods in Molecular Biology, vol 2654. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3135-5_31
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3135-5_31
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3134-8
Online ISBN: 978-1-0716-3135-5
eBook Packages: Springer Protocols