title: Importance of combined treatment with IL-10 and IL-4, but not IL-13, for inhibition of monocyte release of the Ca(2+)-binding protein MRP8/14.
abstract: Expression of the two myeloic related proteins MRP8 and MRP14 is restricted to distinct stages of monocytic differentiation. Heterodimeric MRP8/14 complexes (27E10 antigen) have been shown to represent their biologically active forms. In this study, we investigated the effects of Th2-cytokines on release of these proteins from freshly obtained blood monocytes and monocytes cultured for 7 days in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Monocytes were stimulated with pokeweed mitogen (PWM) in the presence or absence of interleukin-13 (IL-13), IL-4 and IL-10, and secretion of MRP8, MRP14 and MRP8/14 was assessed by using a sandwich enzyme-linked immunosorbent assay system. Peripheral monocytes secreted significantly increased amounts of MRP14 and MRP8/14 but not MRP8 under stimulation with PWM. IL-10 and IL-4, but not IL-13, down-regulated the PWM-stimulated MRP8/14 secretion in a dose-dependent manner. Maximal inhibition required that IL-10 and IL-4 be added up to 1 h before or simultaneous with PWM. A combination of IL-10 and IL-4 even at suboptimal concentrations significantly suppressed protein secretion much more than using IL-10 or IL-4 at a doubled concentration alone. Peripheral monocytes cultured for 7 days in the presence of GM-CSF showed two-to threefold higher protein levels compared with freshly obtained blood monocytes but responded inefficiently to either IL-4, IL-13, or IL-10 alone. However, treatment with IL-10 in combination with IL-4 but not IL-13 strongly suppressed MRP14 and MRP8/14 release by these cells. The unresponsiveness of 7-day-cultured blood macrophages suggests that more differentiated and activated cells may lose their ability to respond to anti-inflammatory cytokines. Combined cytokine treatment may therefore more effectively control the progression of chronic inflammatory processes.
authors.affiliation: Department of Medicine B, University of Münster, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Lügering
authors.forename: T
authors.initials: T
authors.lastname: Kucharzik
authors.forename: A
authors.initials: A
authors.lastname: Lügering
authors.forename: G
authors.initials: G
authors.lastname: Winde
authors.forename: C
authors.initials: C
authors.lastname: Sorg
authors.forename: W
authors.initials: W
authors.lastname: Domschke
authors.forename: R
authors.initials: R
authors.lastname: Stoll
mesh_terms: D000943:Antigens, Differentiation / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D040501:Calgranulin A; D040502:Calgranulin B; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000276:immunology; D006801:Humans; D016753:Interleukin-10 / Q000276:immunology; D018793:Interleukin-13 / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D007378:Interleukins / Q000276:immunology*; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism; D011043:Pokeweed Mitogens / Q000276:immunology
chemical_list: D000943:Antigens, Differentiation; D002135:Calcium-Binding Proteins; D040501:Calgranulin A; D040502:Calgranulin B; D018793:Interleukin-13; D007378:Interleukins; D011043:Pokeweed Mitogens; D016753:Interleukin-10; D015847:Interleukin-4