Extraction of human Langerhans cells: a method for isolation of epidermis-resident dendritic cells

Citation
V. Pena-cruz et al., Extraction of human Langerhans cells: a method for isolation of epidermis-resident dendritic cells, J IMMUNOL M, 255(1-2), 2001, pp. 83-91
Citations number
22
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Immunology
Journal title
JOURNAL OF IMMUNOLOGICAL METHODS
ISSN journal
0022-1759 → ACNP
Volume
255
Issue
1-2
Year of publication
2001
Pages
83 - 91
Database
ISI
SICI code
0022-1759(20010901)255:1-2<83:EOHLCA>2.0.ZU;2-B
Abstract
Langerhans cells (LCs) are immature dendritic cells in the epidermis that p lay a central role in T-lymphocyte mediated skin immunity, Upon activation with antigenic stimuli, they differentiate drastically into mature dendriti c cells while migrating from the epidermis to regional lymph nodes. Thus, i n order to study biological details of immature LCs, it is crucial to isola te epidermis-resident, immature LCs without dermal dendritic cell contamina tion. Methods for extracting LCs from human skin as well as in vitro deriva tion of LC-like cells from hematopoietic progenitor cells have been describ ed previously, but the cell preparations can potentially contain a signific ant number of dendritic cells that are not identical to epidermal LCs. Here , we describe a technique by which purely epidermis-resident LCs are extrac ted from human skin. Following digestion of human skin with dispase, the ep idermis was separated mechanically without any attached dermal component. T he trypsinized epidermal cells were then fractionated by centrifugation wit h a discontinuous density gradient composed of bovine albumin and sodium me trizoate. The LC-enriched preparation thus obtained contained 80% to > 90% CD1a(+), E-cadherin(+) cells that expressed Birbeck granules and the Lag pr otein. Consistent with their being at an immature stage, the freshly isolat ed LCs lacked the expression of CD83, a marker for mature dendritic cells. The purified LCs were able to activate allogeneic T cells, indicating that the cells retained T-cell stimulation ability even after extraction. Thus, the present work offers an opportunity for precise in vitro studies of epid ermal LCs. (C) 2001 Elsevier Science BN. All rights reserved.