Abstract
In the human skin, various types of antigen-presenting cells (APC) are present. In the epidermis, they are identified ultrastructurally as Langerhans cells (LC) by the presence of Birbeck granules. LC are considered to belong to the family of dendritic cells (DC) that are important for the initiation of immune responses (1). In the dermis, macrophages and DC are present (2,3). The expression of CD1a molecules can be used to identify DC in the skin (4,5), because macrophages do not express this marker. In vivo, these skin DC are supposed to take up antigens penetrating in the skin. Thereafter, they migrate via the afferent lymphatics into the draining lymph nodes, where a T-cell response can be initiated (6,7). During migration, the DC mature into potent APC. Besides an increase in MHC class II expression, adhesion (8,9) and B7 co-stimulatory molecules (10) are up-regulated. Most research on skin DC has been carried out with cells isolated from enzyme digested skin (8-10). In this chapter, we describe a method to obtain DC from human skin without enzymes, by making use of their migratory capacities. The cells migrate “spontaneously” out of the skin during culture. Characterization of the cells shows that mature DC are obtained with a marker expression not influenced by enzymes.
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Steinman, R. M. (1990) The dendritic cell system and its role in immunogenicity. Annu. Rev. Immunol. 9, 271–296.
Richters, C. D., Hoekstra, M. J., van Baare, J., Du Pont, J. S., Hoefsmit, E. C., and M., Kamperdijk, E. W. A. (1994) Isolation and characterization of migratory human skin dendritic cells. Clin. Exp. Immunol. 98, 330–336.
Richters, C. D., Hoekstra, M. J., van Baare, J., Du Pont, J. S., Hoefsmit, E. C. M., Kamperdijk, E.W.A. (1995) Migratory properties and functional capacities of human skin dendritic cells. Br. J. Dermatol. 133, 721–727.
Murphy, G. F., Bhan, A. K. Sato, S., Mihm M. C., and Harrist, T. J. (1981) A new immunologic marker for human Langerhans cells. N. Engl. J. Med. 304, 791–792.
Meunier, L., Gonzalez-Ramos, A., and Cooper K.D. (1993) Heterogeneous populations of class II MHC+cells in human dermal cell suspensions. Identification of a small subset responsible for potent dermal antigen-presenting cell activity with features analogues to Langerhans cells. J. Immunol. 151, 4067–4080.
Silberberg-Sinakin, I., Thorbecke, G. J., Baer, R. L., and Berezowsky, V. (1976) Antigen bearing Langerhans cells in skin, dermal lymphatics and in lymph nodes. Cell Immunol. 25, 137–141.
Hoefsmit, E. C. M., Duijvestijn, A. M., and Kamperdijk, E. W. A. (1982) Relation between Langerhans cells, veiled cells and interdigitating cells. Immunobiol. 161, 225–265.
Teunissen, M. B. M., Wormmeester, J., Krieg, S. R.,et al. (1990) Human epidermal Langerhans cells undergo profound morphological and phenotypical changes during in vitro culture. J. Invest. Dermatol. 94, 166–173.
Romani, N., Lenz, A., Glessel, H., et al. (1989) Cultured Langerhans cells resemble lymphoid dendritic cells in phenotype and function. J. Invest. Dermatol., 93, 600–609.
Symington, F. W., Brady, W., and Linsley, P. S. (1993) Expression and function of B7 on human epidermal Langerhans cells. J. Immunol. 150, 1286–1295.
Cordell, J. L., Faline, B., Erber, W. N., et al. (1984) Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J. Histochem. Cytochem. 32, 219–229.
Pease, D. C. (ed.) (1964) Histological techniques for electron microscopy. Academic Press, New York.
Liou, W., Geuze, H. J., and Slot, J. W. (1996) Improving structural integrity of cryosections for immunogold labeling. Histochem. Cell. Biol. 106, 41–58.
Sallusto, F., Cella, M., Danielli, C., and Lanzavecchia, A. (1995) Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the MHC class II compartment: downregulation by cytokines and bacterial products. J. Exp. Med. 182, 389–400.
Witmer-Pack, M. D., Olivier W., Valinsky, J., Schuler, G., and Steinman, R. M. (1987) Granulocyte/macrophage colony stimulating factor is essential for the viability and function of cultured murine epidermal Langerhans cells. J. Exp. Med. 166, 1484–1498.
Heufler, C., Koch, F., and Schuler, G. (1988) Granulocyte/macrophage colonystimulating factor and interleukin 1 mediate the maturation of murine epidermal Langerhans cells into potent immunostimulatory dendritic cells. J. Exp. Med. 167, 700–705.
Richters, C. D., Hoekstra, M. J., Hoefsmit, E. C. M., Kamperdijk, E. W. A. (1995) Phenotype of cells migrated from human skin explants. Adv. Exp. Med. Biol. 378, 247–251.
Richters, C. D., Reits, E. A. J., van Pelt, A. M., et al. (1996) Effect of low dose UVB irradiation on the migratory properties and functional capacities of human skin dendritic cells. Clin. Exp. Immunol. 104, 191–197.
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Richters, C.D., Hoekstra, M.J., du Pont, J.S., Kreis, R.W., Kamperdijk, E.W.A. (2001). Isolation of Human Skin Dendritic Cells by In Vitro Migration. In: Robinson, S.P., Stagg, A.J., Knight, S.C. (eds) Dendritic Cell Protocols. Methods in Molecular Medicine™, vol 64. Humana Press. https://doi.org/10.1385/1-59259-150-7:145
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DOI: https://doi.org/10.1385/1-59259-150-7:145
Publisher Name: Humana Press
Print ISBN: 978-0-89603-584-3
Online ISBN: 978-1-59259-150-3
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