title: Bacterial lipopolysaccharide, TNF-alpha, and calcium ionophore under serum-free conditions promote rapid dendritic cell-like differentiation in CD14+ monocytes through distinct pathways that activate NK-kappa B.
abstract: 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-alpha, 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-alpha 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-kappaB family of transcription factors, including RelB, suggesting that despite differences in upstream elements, the signaling pathways all involve NF-kappaB. 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.
authors.affiliation: Division of Basic Sciences, Frederick Cancer Research and Development Center, National Cancer Institute, MD 21702, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Lyakh
authors.forename: G K
authors.initials: GK
authors.lastname: Koski
authors.forename: W
authors.initials: W
authors.lastname: Telford
authors.forename: R E
authors.initials: RE
authors.lastname: Gress
authors.forename: P A
authors.initials: PA
authors.lastname: Cohen
authors.forename: N R
authors.initials: NR
authors.lastname: Rice
mesh_terms: D000017:ABO Blood-Group System / Q000276:immunology; D000595:Amino Acid Sequence; D015703:Antigens, CD; D000001:Calcimycin / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D002467:Cell Nucleus / Q000276:immunology / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins / Q000502:physiology; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D006131:Growth Inhibitors / Q000276:immunology; D006133:Growth Substances / Q000502:physiology; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D007136:Immunoglobulins / Q000096:biosynthesis; D016130:Immunophenotyping; D007476:Ionophores / Q000494:pharmacology; D007958:Leukocyte Count; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis*; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D016328:NF-kappa B / Q000096:biosynthesis / Q000378:metabolism* / Q000502:physiology; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D020033:Protein Isoforms / Q000096:biosynthesis; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D015398:Signal Transduction / Q000276:immunology*; D051841:Transcription Factor RelB; D014157:Transcription Factors / Q000096:biosynthesis / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000276:immunology / Q000494:pharmacology*; D000097802:CD83 Antigen
chemical_list: D000017:ABO Blood-Group System; D015703:Antigens, CD; D000001:Calcimycin; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins; D006131:Growth Inhibitors; D006133:Growth Substances; D007106:Immune Sera; D007136:Immunoglobulins; D007476:Ionophores; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D020033:Protein Isoforms; D011518:Proto-Oncogene Proteins; D051841:Transcription Factor RelB; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D000097802:CD83 Antigen; C497593:RELB protein, human