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Clinical Research Unit, Department of Dermatology, Johannes Gutenberg University, Mainz, Germany
Dendritic cells (DC) are characterized by their unique potential to
prime naive T cells. Epidermal Langerhans cells (LC), the DC resident
in the epidermis, gain this immunostimulatory capacity following Ag
contact in vivo or during in vitro culture of epidermal cell
suspensions. To analyze differential gene expression in maturing LC, we
constructed a highly representative cDNA library of cultivated LC (cLC)
in 
ZAP II containing 18 x 106 independent
clones. This library was screened with freshly isolated Langerhans cell
(fLC)- and cLC-derived probes for cLC-specific cDNAs. The cDNAs
identified were sequenced and analyzed by database searches. Two cDNA
fragments were identified as fragments of fascin, indicating that
fascin is differentially expressed in LC. By competitive RT-PCR, we
confirmed that fascin is highly expressed in cLC cultivated for 1, 2,
and 3 days, while no signals were obtained with fLC. Western blot and
immunofluorescence analysis revealed cLC-specific expression of fascin
on the protein level as well. Fascin is known to be involved in the
organization of the actin cytoskeleton in cytoplasmatic extensions of
nerve growth cones. Its differential expression in maturing LC
coincides with the formation of numerous dendritic projections in LC.
Their formation was inhibited by incubation of LC with fascin antisense
oligonucleotides during cultivation. Therefore, we conclude that fascin
is necessary for the formation of the dendritic processes of maturing
Langerhans cells and may thus influence T cell-LC interaction.
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