Document Detail


CD1a and CD1c cell sorting yields a homogeneous population of immature human Langerhans cells.
MedLine Citation:
PMID:  12969546     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
There is increasing evidence that ex vivo generated Langerhans cells (LCs) cannot fully substitute for their physiological counterparts in normal epidermis when studying the immunobiology of this prototype of a tissue-residing immature dendritic cell (DC). Here, we present CD1-based magnetic-activated cell-sorting (MACS) protocols for the effective isolation of human epidermal LCs. CD1c selection yielded a homogeneous population of pure and viable HLA-DR(+)/CD1a(+) DCs, with the ultrastructural features, surface antigen expression and cytokine profile, characteristic of epidermis-resident immature LCs. The immature state and functional integrity were established by allogeneic mixed lymphocyte reactions showing a weak stimulatory capacity of freshly isolated cells and upregulation upon stimulation. Characterizing the cells in more detail, we could demonstrate for the first time that normal human LCs express CXCR4, CD40 ligand (CD40L), and Fas and Fas ligand (FasL). The observed constitutive transcription of TGF-beta suggests that the viability and immature state of epidermal LCs are maintained not only by the TGF-beta production from the microenvironment, but also in an autocrine or paracrine manner. LPS and IFN-omega stimulated the expression of the inflammatory cytokines TNF-alpha and IL-1beta, and there was secretion of IL-12p70 after CD40 ligation. Remarkably, the CD1-sorted LCs showed no loss of their Birbeck granules and CD1a expression upon culturing and no spontaneous phenotypic and functional maturation into potent antigen-presenting cells (APCs). We conclude that human epidermal LCs obtained by the CD1c cell-sorting protocol are optimal candidates with which to elucidate the properties and capabilities of immature cells and to develop immunotherapeutic vaccines.
Authors:
Matthias Peiser; Andreas Grützkau; Reinhard Wanner; Gerhard Kolde
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of immunological methods     Volume:  279     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  2003 Aug 
Date Detail:
Created Date:  2003-09-12     Completed Date:  2003-11-04     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  41-53     Citation Subset:  IM    
Affiliation:
Department of Dermatology and Allergy, Charité, Humboldt-University of Berlin, D-10117 Berlin, Germany.
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MeSH Terms
Descriptor/Qualifier:
Antigens, CD
Antigens, CD1 / immunology*
Antigens, CD80 / immunology
Flow Cytometry / methods*
HLA-DR Antigens / immunology
Humans
Immunoglobulins / immunology
Langerhans Cells / cytology,  immunology*,  ultrastructure
Membrane Glycoproteins / immunology
Microscopy, Electron
Skin / cytology,  immunology
Chemical
Reg. No./Substance:
0/Antigens, CD; 0/Antigens, CD1; 0/Antigens, CD80; 0/CD83 antigen; 0/HLA-DR Antigens; 0/Immunoglobulins; 0/Membrane Glycoproteins

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