title: Calcium mobilization in human myeloid cells results in acquisition of individual dendritic cell-like characteristics through discrete signaling pathways.
abstract: We have shown previously that calcium ionophore (CI) treatment of various myeloid origin cells results in rapid acquisition of properties associated with mature, activated dendritic cells. These properties include increased CD83 and costimulatory molecule expression, tendencies to form dendritic processes, loss of CD14 expression by monocytes, and typically an enhanced capacity to sensitize T lymphocytes to Ag. We here analyze the intracellular signaling pathways by which CI induces acquisition of such properties. Thapsigargin, which raises intracellular Ca2+ levels by antagonizing its sequestration, induced immunophenotypic and morphologic changes that paralleled CI treatment. CI-induced activation was broadly attenuated by the Ca2+ chelating compound EGTA and by calmodulin antagonists trifluoperazine dimaleate and W-7. However, antagonists of signaling pathways downstream to calmodulin displayed more selective inhibitory effects. Calcineurin antagonists cyclosporin A and the FK-506 analogue, ascomycin, diminished costimulatory molecule and CD83 expression, as well as formation of dendritic processes in CI-treated myeloid cells, and strongly attenuated the T cell allosensitizing capacity of CI-treated HL-60 cells. These calcineurin antagonists displayed minimal effect on CI-induced CD14 down-regulation in monocytes. In contrast, the calmodulin-dependent protein kinase antagonists, K252a and KT5926, while displaying only modest effects on CI-induced costimulatory molecule and CD83 expression, strongly blocked CD14 down-regulation. These results are consistent with a Ca2+-dependent mechanism for CI-induced differentiation of myeloid cells, and indicate that multiple discrete signaling pathways downstream to calcium mobilization and calmodulin activation may be essential in regulating this process.
authors.affiliation: Medicine Branch, National Cancer Institute, Bethesda, MD 20892, USA. gkoski@box-g.nih.gov
authors.forename: G K
authors.initials: GK
authors.lastname: Koski
authors.forename: G N
authors.initials: GN
authors.lastname: Schwartz
authors.forename: D E
authors.initials: DE
authors.lastname: Weng
authors.forename: B J
authors.initials: BJ
authors.lastname: Czerniecki
authors.forename: C
authors.initials: C
authors.lastname: Carter
authors.forename: R E
authors.initials: RE
authors.lastname: Gress
authors.forename: P A
authors.initials: PA
authors.lastname: Cohen
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis; D018952:Antigens, CD34 / Q000032:analysis; D065095:Calcineurin Inhibitors; D020013:Calcium Signaling / Q000276:immunology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D003713:Dendritic Cells / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D015536:Down-Regulation / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018922:HL-60 Cells / Q000187:drug effects / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000096:biosynthesis / Q000494:pharmacology; D016130:Immunophenotyping; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D016559:Tacrolimus / Q000031:analogs & derivatives / Q000494:pharmacology; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D065095:Calcineurin Inhibitors; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D016572:Cyclosporine; D004791:Enzyme Inhibitors; D007136:Immunoglobulins; D007166:Immunosuppressive Agents; D007476:Ionophores; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D016559:Tacrolimus; D000097802:CD83 Antigen; C058028:immunomycin