Cultured human Langerhans cells resemble lymphoid dendritic cells in phenotype and function.
Freshly isolated murine epidermal Langerhans cells (LC) are weak stimulators of resting T cells. Upon culture their phenotype changes, their stimulatory activity increases significantly, and they come to resemble lymphoid dendritic cells. Resident murine LC, therefore, might represent a reservoir of immature dendritic cells. We have now used enzyme cytochemistry, a panel of some 80 monoclonal antibodies, and immunofluorescence microscopy or two-color flow cytometry, as well as transmission electron microscopy, to analyse the phenotype and morphology of human LC before and after 2-4 d of bulk epidermal cell culture. In addition, LC were enriched from bulk epidermal cell cultures, and their stimulatory capacity was tested in the allogeneic mixed leukocyte reaction and the oxidative mitogenesis assay. Cultured human LC resembled human lymphoid dendritic cells in morphology, phenotype, and function. Specifically, LC became non-adherent upon culture and developed sheet-like processes (so-called "veils"), decreased their surface ATP/ADP'ase activity, and lost nonspecific esterase activity. As in the mouse, surface expression of MHC class I and II antigens increased significantly, and FcII receptors were significantly reduced. Markers that are expressed by dendritic cells (like CD40) appeared on LC following culture. Cultured human LC were potent T-cell stimulators. Our findings support the view that resident human LC, like murine LC, represent immature precursors of lymphoid dendritic cells in skin-draining lymph nodes.
Romani
N
N
Department of Dermatology, University of Innsbruck, Austria.
Lenz
A
A
Glassel
H
H
Stössel
H
H
Stanzl
U
U
Majdic
O
O
Fritsch
P
P
Schuler
G
G
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000954:Antigens, Surface / Q000032:analysis; D015815:Cell Adhesion Molecules / Q000032:analysis; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006649:Histocompatibility Antigens; D006801:Humans; D066298:In Vitro Techniques; D007801:Langerhans Cells / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D017493:Leukocyte Common Antigens; D008213:Lymphocyte Activation; D008285:Major Histocompatibility Complex; D008854:Microscopy, Electron; D011961:Receptors, Fc / Q000032:analysis
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D006649:Histocompatibility Antigens; D011961:Receptors, Fc; D017493:Leukocyte Common Antigens