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, and Calcium Ionophore Under Serum-Free Conditions Promote Rapid Dendritic Cell-Like Differentiation in CD14+ Monocytes Through Distinct Pathways That Activate NF-
B
*
Division of Basic Sciences, Frederick Cancer Research and Development Center, National Cancer Institute, Frederick, MD 21702;
Medicine Branch, National Cancer Institute, Bethesda, MD 20814; and
The Center for Surgery Research, Cleveland Clinic Foundation, Cleveland, OH 44195
To facilitate the study of signaling pathways involved in myeloid
dendritic cell (DC) differentiation, we have developed a serum-free
culture system in which human CD14+ peripheral blood
monocytes differentiate rapidly in response to bacterial LPS, TNF-
,
or calcium ionophore (CI). Within 48–96 h, depending on the inducing
agent, the cells acquire many immunophenotypical, morphological,
functional, and molecular properties of DC. However, there are
significant differences in the signaling pathways used by these agents,
because 1) LPS-induced, but not CI-induced, DC differentiation required
TNF- production; and 2) cyclosporin A inhibited differentiation
induced by CI, but not that induced by LPS. Nevertheless, all three
inducing agents activated members of the NF-
B family of
transcription factors, including RelB, suggesting that despite
differences in upstream elements, the signaling pathways all involve
NF-B. In this report we also demonstrate and offer an explanation
for two observed forms of the RelB protein and show that RelB can be
induced in myeloid cells, either directly or indirectly, through a
calcium-dependent and cyclosporin A-sensitive
pathway.
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