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,§
*
Department of Cell Biology and Ludwig Institute for Cancer Research,
Department of Laboratory Medicine,
Molecular Cardiobiology Program, and
§
Section of Immunobiology, Yale University School of Medicine, New Haven, CT 06520
Dendritic cells (DCs) play a critical role as APCs in the induction
of the primary immune response. Their capacity for Ag processing and
presentation is tightly regulated, controlled by a terminal
developmental sequence accompanied by striking changes in morphology,
organization, and function. The maturation process, which converts DCs
from cells adapted for Ag accumulation to cells adapted for T cell
stimulation, remains poorly understood due in part to difficulties in
the culture and manipulation of DCs of defined lineages. To address
these issues, we have devised conditions for the culture of a single DC
type, Langerhans cells (LCs), using CD34+ cells from
G-CSF-mobilized patients. Homogenous populations of LCs, replete with
abundant immunocytochemically demonstrable Birbeck granules, could be
stably maintained as immature DCs for long periods in culture. Unlike
other human DC preparations, the LCs remained fully differentiated
after cytokine removal. Following exposure to TNF-
, LPS, or CD40
ligand, the LCs could be synchronously induced to mature. Depending on
the agent used, distinct types of LCs emerged differing in their
capacity for T cell stimulation, IL-12 production, intracellular
localization of MHC products, and overall morphology. Most
interestingly, the expression of different sets of Toll family
receptors is induced or down-regulated according to the maturation
stimulus provided. These results strongly suggest that different
proinflammatory stimuli might drive distinct developmental
events.
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