Abstract
Skin dendritic cells (DC) are strategically positioned at the body’s second largest epithelial border to the environment. Hence they are the first antigen presenting cells that encounter invading pathogens and environmental antigens, including contact sensitizers and carcinogens penetrating the skin. Moreover, DC have the unique ability to induce immunity or tolerance and thus take center stage in regulating innate and adaptive immune responses. Skin DC can be divided into several phenotypically and functionally distinct subtypes. The three main subsets are Langerin+ epidermal Langerhans cells (LC) and Langerin+ as well as Langerinneg dermal DC. In the steady state skin DC form a dense network to survey the periphery for pathogens and harmful substances breaching the cutaneous barrier. During inflammation DC become rapidly activated and start their migration to skin-draining lymph nodes where they initiate antigen-specific T cell responses. The homeostasis and mobilization of DC in the skin can be visualized by immunofluorescent staining of epidermal and dermal sheet preparations or skin sections. Here, we describe in detail how inflammation can be induced in the skin with tape stripping or FITC painting and how the skin DC network can be monitored using immunofluorescence microscopy and flow cytometry.
Note: In the printed book and online version of chapter 3, an asterisk (*) was mistakenly omitted after the author Patrizia Stoitzner’s name. It has now been inserted to show that she and Björn E. Clausen have contributed equally to this chapter.
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References
Clausen BE, Stoitzner P (2015) Functional specialization of skin dendritic cell subsets in regulating T cell responses. Front Immunol 6:534
Romani N, Clausen BE, Stoitzner P (2010) Langerhans cells and more: langerin-expressing dendritic cell subsets in the skin. Immunol Rev 234:120–41
Jiang A, Bloom O, Ono S, Cui W, Unternaehrer J, Jiang S, Whitney JA, Connolly J, Banchereau J, Mellman I (2007) Disruption of E-cadherin-mediated adhesion induces a functionally distinct pathway of dendritic cell maturation. Immunity 27:610–24
Steinman RM, Hawiger D, Nussenzweig MC (2003) Tolerogenic dendritic cells. Annu Rev Immunol 21:685–711
Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392:245–52
Dalod M, Chelbi R, Malissen B, Lawrence T (2014) Dendritic cell maturation: functional specialization through signaling specificity and transcriptional programming. EMBO J 33:1104–16
Steinman RM, Banchereau J (2007) Taking dendritic cells into medicine. Nature 449:419–26
Henri S, Poulin LF, Tamoutounour S, Ardouin L, Guilliams M, de Bovis B, Devilard E, Viret C, Azukizawa H, Kissenpfennig A, Malissen B (2010) CD207+ CD103+ dermal dendritic cells cross-present keratinocyte-derived antigens irrespective of the presence of langerhans cells. J Exp Med 207:189–206
Tamoutounour S, Guilliams M, Montanana Sanchis F, Liu H, Terhorst D, Malosse C, Pollet E, Ardouin L, Luche H, Sanchez C, Dalod M, Malissen B, Henri S (2013) Origins and functional specialization of macrophages and of conventional and monocyte-derived dendritic cells in mouse skin. Immunity 39:925–38
Geissmann F, Manz MG, Jung S, Sieweke MH, Merad M, Ley K (2010) Development of monocytes, macrophages, and dendritic cells. Science 327:656–61
Malissen B, Tamoutounour S, Henri S (2014) The origins and functions of dendritic cells and macrophages in the skin. Nat Rev Immunol 14:417–28
Seré K, Baek JH, Ober-Blöbaum J, Müller-Newen G, Tacke F, Yokota Y, Zenke M, Hieronymus T (2012) Two distinct types of Langerhans cells populate the skin during steady state and inflammation. Immunity 37:905–16
Pulendran B, Tang H, Denning TL (2008) Division of labor, plasticity, and crosstalk between dendritic cell subsets. Curr Opin Immunol 20:61–7
Honda T, Nakajima S, Egawa G, Ogasawara K, Malissen B, Miyachi Y, Kabashima K (2010) Compensatory role of Langerhans cells and langerin-positive dermal dendritic cells in the sensitization phase of murine contact hypersensitivity. J Allergy Clin Immunol 125:1154–1156.e2
Noordegraaf M, Flacher V, Stoitzner P, Clausen BE (2010) Functional redundancy of Langerhans cells and Langerin + dermal dendritic cells in contact hypersensitivity. J Invest Dermatol 130:2752–9
Kautz-Neu K, Noordegraaf M, Dinges S, Bennett CL, John D, Clausen BE, von Stebut E (2011) Langerhans cells are negative regulators of the anti-Leishmania response. J Exp Med 208:885–91
Igyártó BZ, Haley K, Ortner D, Bobr A, Gerami-Nejad M, Edelson BT, Zurawski SM, Malissen B, Zurawski G, Berman J, Kaplan DH (2011) Skin-resident murine dendritic cell subsets promote distinct and opposing antigen-specific T helper cell responses. Immunity 35:260–72
Flacher V, Tripp CH, Mairhofer DG, Steinman RM, Stoitzner P, Idoyaga J, Romani N (2014) Murine Langerin + dermal dendritic cells prime CD8+ T cells while Langerhans cells induce cross-tolerance. EMBO Mol Med 6:1191–204
Holzmann S, Tripp CH, Schmuth M, Janke K, Koch F, Saeland S, Stoitzner P, Romani N (2004) A model system using tape stripping for characterization of Langerhans cell-precursors in vivo. J Invest Dermatol 122:1165–74
Juhlin L, Shelley WB (1977) New staining techniques for the Langerhans cell. Acta Derm Venereol 57:289–96
Lessard RJ, Wolff K, Winkelmann RK (1968) The disappearance and regeneration of Langerhans cells following epidermal injury. J Invest Dermatol 50:171–9
Streilein JW, Lonsberry LW, Bergstresser PR (1982) Depletion of epidermal langerhans cells and Ia immunogenicity from tape-stripped mouse skin. J Exp Med 155:863–71
Wood LC, Jackson SM, Elias PM, Grunfeld C, Feingold KR (1992) Cutaneous barrier perturbation stimulates cytokine production in the epidermis of mice. J Clin Invest 90:482–7
Stoitzner P, Zanella M, Ortner U, Lukas M, Tagwerker A, Janke K, Lutz MB, Schuler G, Echtenacher B, Ryffel B, Koch F, Romani N (1999) Migration of langerhans cells and dermal dendritic cells in skin organ cultures: augmentation by TNF-alpha and IL-1beta. J Leukoc Biol 66:462–70
Acknowledgements
The authors would like to thank the members of the Clausen and the Stoitzner laboratories for their support and the members of the animal facilities of the University Medical Center Mainz and the Medical University of Innsbruck for responsible animal husbandry. J.L.O. is a fellow of the Fritz-Thyssen Foundation (1O.15.1.O2OMN), D.O. is supported by a scholarship from the Austrian Science Fund (FWF-T-007370), B.E.C. is supported by the DFG (CL 419/2-1) and the Research Center for Immunotherapy (FZI) Mainz, and P.S. is supported by grants form the Austrian Science Fund (P-27001-B13, ZFW011010-015).
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Ober-Blöbaum, J.L. et al. (2017). Monitoring Skin Dendritic Cells in Steady State and Inflammation by Immunofluorescence Microscopy and Flow Cytometry. In: Clausen, B., Laman, J. (eds) Inflammation. Methods in Molecular Biology, vol 1559. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6786-5_3
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DOI: https://doi.org/10.1007/978-1-4939-6786-5_3
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