title: Prostaglandin E2 secretion, cell maturation, and CD14 expression by monocyte-derived macrophages from localized juvenile periodontitis patients.
abstract: Previous studies from our laboratory have demonstrated that freshly isolated peripheral blood monocytes from localized juvenile periodontitis (LJP) patients secrete more prostaglandin E2 (PGE2) after stimulation by lipopolysaccharide (LPS) than do monocytes from healthy subjects. However, it is not clear if the altered function of LJP monocytes is intrinsic to the cells or is induced by the persistent infection of the periodontium. The present study was designed to compare PGE2 secretion by freshly-isolated peripheral blood monocytes (FIM) from LJP and control subjects to in vitro monocyte-derived macrophages (MDM) from the same subjects. We also examined monocyte maturation into macrophage-like cells and the cell-surface expression of the LPS, receptor, CD14 during the culture period. FIM from LJP patients and controls were stimulated by different concentrations of LPS (O to 30 micrograms/ml) for 24 hours. These experiments were performed immediately after cell separation and after 10 days in culture, which allowed differentiation of monocytes into MDM. PGE2 levels in the culture media were determined using a radioimmunoassay. Cell surface expression of CD71, a cell maturation marker, and CD14 were assayed by cell-ELISA in relation to beta-2-microglobulin. LPS-stimulated FIM from LJP patients secreted 3 to 4 times more PGE2 than control FIM at all LPS concentrations tested. After 10 days in culture, the LJP MDM secretion of PGE2 reduced to control MDM level of PGE2 secretion. These levels of PGE2 secretion were comparable to PGE2 secretion from FIM of controls. Cell maturation, as verified by CD71 expression, was found not to differ between the groups. However, the expression of CD14 by LJP FIM was much lower than on control FIM (approximately equal to 50%). After 5 or 10 days in culture, MDM from both control and LJP subjects expressed comparable amounts of CD14. The results suggest that in vitro conditions reverse the hypersensitivity of LJP monocytes to LPS into control levels and CD14 expression is not correlated to the hyper-responsiveness of the cells to LPS.
authors.affiliation: Department of Periodontology, Hebrew University-Hadassah Faculty of Dental Medicine, Jerusalem, Israel.
authors.forename: L
authors.initials: L
authors.lastname: Shapira
authors.forename: W A
authors.initials: WA
authors.lastname: Soskolne
authors.forename: T E
authors.initials: TE
authors.lastname: Van Dyke
mesh_terms: D000293:Adolescent; D010520:Aggressive Periodontitis / Q000276:immunology* / Q000473:pathology; D015703:Antigens, CD / Q000032:analysis / Q000235:genetics; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis / Q000235:genetics; D016022:Case-Control Studies; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D015232:Dinoprostone / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology; D008264:Macrophages / Q000276:immunology*; D008297:Male; D009000:Monocytes / Q000276:immunology*; D011990:Receptors, Transferrin; D001613:beta 2-Microglobulin / Q000032:analysis
chemical_list: D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D011990:Receptors, Transferrin; D001613:beta 2-Microglobulin; D015232:Dinoprostone