Abstract
Studies regarding cellular interactions between Langerhans cells and other skin cells are somehow hampered by the difficult cultivation of these cells in vitro. Here, we show that the human MUTZ-3 cell line can be differentiated into Langerhans-like cells in the presence of a cytokine cocktail including GM-CSF, TGF-β1 and TNF-α. We used the expression of langerin, CD1a, CCR6 and the intracellular presence of Birbeck granules to identify the differentiated MUTZ-3 cells (MUTZ-3-LCs). The aim of this study was to integrate MUTZ-3-LCs into a three-dimensional full-thickness skin model. On top of fibroblast-containing collagen matrix a mixture of primary human keratinocytes and MUTZ-3-LCs were seeded and cultured for 24 h. Subsequently, the models were lifted up to the air-liquid interface. Histological evaluation featured a fully stratified epidermis with all characteristic epidermal strata. Langerin-positive cells were detected suprabasally within the epidermis indicating that keratinocytes provide environmental conditions for long-time maintenance of MUTZ-3-LCs. These skin models provide a tool to further investigate the interactions between Langerhans-like cells and other skin cells and particularly learn more about the cutaneous immune response.
References
Birbeck MS, Breathnach AS, Everall JD (1961) An electron microscope study of basal melanocytes and high-level clear cells (Langerhans cells) in vitiligo. J Invest Dermatol 37:51
Borkowski TA, Letterio JJ, Farr AG, Udey MC (1996) A role for endogenous transforming growth factor β1 in Langerhans cell biology: the skin of transforming growth factor β1 null mice is devoid of epidermal Langerhans cells. J Exp Med 184:2417–2422
Caux C, Dezutter-Dambuyant C, Schmitt D, Banchereau J (1992) GM-CSF and TNF-alpha cooperate in the generation of dendritic Langerhans cells. Nature 360:258–261
Charbonnier AS, Kohrgruber N, Kriehuber E, Stingl G, Rot A, Maurer D (1999) Macrophage inflammatory protein 3α is involved in the constitutive trafficking of epidermal Langerhans cells. J Exp Med 190:1755–1768
Cumberbatch M, Dearman RJ, Griffiths CE, Kimber I (2000) Langerhans cell migration. Clin Exp Dermatol 25:413–418
Geissmann F, Prost C, Monnet JP, Dy M, Brousse N, Hermine O (1998) Transforming growth factor b1 in the presence of granulocyte/macrophage colony-stimulating factor and interleukin 4, induces differentiation of human peripheral blood monocytes into dendritic Langerhans cells. J Exp Med 187:961–966
Kaplan DH, Li MO, Jenison MC, Shlomchik WD, Flavell RA, Shlomchik MJ (2007) Autocrine/paracrine TGFbeta1 is required for the development of epidermal Langerhans cells. J Exp Med 204:2545–2552
Kripke ML, Munn CG, Jeevan A, Tang JM, Bucana C (1990) Evidence that cutaneous antigen-presenting cells migrate to regional lymph nodes during contact sensitization. J Immunol 145:2833–2838
Masterson AJ, Sombroek CC, de Gruijl TD, Graus YM, van der Vliet H, Lougheed SM, van den Eertwegh AJ, Pinedo HM, Scheper RJ (2002) MUTZ-3, a human cell line model for the cytokine-induced differentiation of dendritic cells from CD34+ precursors. Blood 100:701–703
Mewes KR, Raus M, Bernd A, Zöller NN, Sättler A, Graf R (2007) Elastin expression in a newly developed full-thickness skin equivalent. Skin Pharmacol Physiol 20:85–95
Pruniéras M, Regnier M, Woodley D (1983) Methods for cultivation of keratinocytes with an air-liquid interface. J Invest Dermatol 81:28s–33s
Reid CD, Stackpoole A, Meager A, Tikerpae J (1992) Interactions of tumor necrosis factor with granulocyte-macrophage colony-stimulating factor and other cytokines in the regulation of dendritic cell growth in vitro from early bipotent CD34+ progenitors in human bone marrow. J Immunol 149:2681–2688
Romani N, Holzmann S, Tripp CH, Koch F, Stoitzner PS (2003) Langerhans cells-dendritic cells of the epidermis. APMIS 111:725–740
Rosdy M, Clauss LC (1990) Terminal epidermal differentiation of human keratinocytes grown in chemically defined medium on inert filter substrates at the air-liquid interface. J Invest Dermatol 95:409–414
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 179:1109–1118
Santegoets SJ, Masterson AJ, van der Sluis PC, Lougheed SM, Fluitsma DM, van den Eertwegh AJ, Pinedo HM, Scheper RJ, de Gruijl TD (2006) A CD34+ human cell line model of myeloid dendritic cell differentiation: evidence for a CD14+CD11b+ Langerhans cell precursor. J Leukoc Biol 80:1337–1344
Strunk D, Egger C, Leitner G et al (1997) A skin homing molecule defines the Langerhans cell progenitor in human peripheral blood. J Exp Med 185:1131–1136
Valladeau J, Clair-Moninot V, Dezutter-Dambuyant C et al (2002) Identification of mouse langerin/CD207 in Langerhans cells and some dendritic cells of lymphoid tissues. J Immunol 168:782–792
Wolff K (1967) The fine structure of the Langerhans cell granule. J Cell Biol 35:468–473
Zöller NN, Kippenberger S, Thaçi D, Mewes K, Spiegel M, Sättler A, Schultz M, Bereiter-Hahn J, Kaufmann R, Bernd A (2008) Evaluation of beneficial and adverse effects of glucocorticoids on a newly developed full-thickness skin model. Toxicol In Vitro 22:747–759
Acknowledgments
We are grateful to Dr. Karsten Mewes and Dr. Lars Vierkotten (Henkel AG & Co. KGaA, Düsseldorf, Germany) for helping us making the collagen matrix. We also thank Dr. Stefan Gfrörer (Paediatric Surgery, University Hospital, Frankfurt/Main, Germany) for providing the foreskin biopsies. This work was in part supported by the Federal Ministry of Education and Research (Bundesministerium für Bildung und Forschung, BMBF; grant-no: 0315018A).
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Laubach, V., Zöller, N., Rossberg, M. et al. Integration of Langerhans-like cells into a human skin equivalent. Arch Dermatol Res 303, 135–139 (2011). https://doi.org/10.1007/s00403-010-1092-x
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DOI: https://doi.org/10.1007/s00403-010-1092-x