title: Characterization of osteoclast precursors in human blood.
abstract: Osteoclast precursors (OCPs) circulate in the mononuclear fraction of peripheral blood (PB), but their abundance and surface characteristics are unknown. Previous studies suggest that the receptor activator for NF-kappaB (RANK) on cytokine-treated OCPs in mouse bone marrow interacts with osteoprotegerin ligand (OPGL/TRANCE/RANKL/ODF) to initiate osteoclast differentiation. Hence, we used a fluorescent form of human OPGL (Hu-OPGL-F) to identify possible RANK-expressing OCPs in untreated peripheral blood mononuclear cells (PBMCs) using fluorescence-activated cell sorting analysis. Monocytes [CD14-phycoerythrin (PE) antibody (Ab) positive (+) cells, 10-15% of PBMCs] all (98-100%) co-labelled with Hu-OPGL-F (n > 18). T lymphocytes (CD3-PE Ab+ cells, 66% of PBMCs) did not bind Hu-OPGL-F; however, B cells (CD19-PE Ab+ cells, 9% of PBMCs) were also positive for Hu-OPGL-F. All Hu-OPGL-F+ monocytes also co-labelled with CD33, CD61, CD11b, CD38, CD45 and CD54 Abs, but not CD34 or CD56 Abs. Hu-OPGL-F binding was dose dependent and competed with excess Hu-OPGL. When Hu-OPGL-F+, CD14-PE Ab+, CD33-PE Ab+, Hu-OPGL-F+/CD14-PE Ab+ or Hu-OPGL-F+/CD33-PE Ab+ cells were cultured with OPGL (20 ng/ml) and colony-stimulating factor (CSF)-1 (25 ng/ml), OC-like cells readily developed. Thus, all freshly isolated monocytes demonstrate displaceable Hu-OPGL-F binding, suggesting the presence of RANK on OCPs in PB; also, OCPs within a purified PB monocyte population form osteoclast-like cells in the complete absence of other cell types in OPGL and CSF-1 containing medium.
authors: Department of Pharmacology/Pathology, Amgen Inc., Thousand Oaks, CA 91320, USA. shalhoub@amgen.com
authors: V
authors: V
authors: Shalhoub
authors: G
authors: G
authors: Elliott
authors: L
authors: L
authors: Chiu
authors: R
authors: R
authors: Manoukian
authors: M
authors: M
authors: Kelley
authors: N
authors: N
authors: Hawkins
authors: E
authors: E
authors: Davy
authors: G
authors: G
authors: Shimamoto
authors: J
authors: J
authors: Beck
authors: S A
authors: SA
authors: Kaufman
authors: G
authors: G
authors: Van
authors: S
authors: S
authors: Scully
authors: M
authors: M
authors: Qi
authors: M
authors: M
authors: Grisanti
authors: C
authors: C
authors: Dunstan
authors: W J
authors: WJ
authors: Boyle
authors: D L
authors: DL
authors: Lacey
mesh_terms: D001402:B-Lymphocytes / Q000378:metabolism; D002352:Carrier Proteins; D002454:Cell Differentiation; D002478:Cells, Cultured; D003115:Colony-Stimulating Factors / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism* / Q000648:ultrastructure; D008562:Membrane Glycoproteins; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D009000:Monocytes / Q000378:metabolism; D010010:Osteoclasts / Q000378:metabolism* / Q000648:ultrastructure; D053244:Osteoprotegerin; D011485:Protein Binding; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D003115:Colony-Stimulating Factors; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D053244:Osteoprotegerin; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C505686:TNFRSF11A protein, human; C506030:TNFRSF11B protein, human; C505688:TNFSF11 protein, human; C505687:Tnfrsf11a protein, mouse; C506031:Tnfrsf11b protein, mouse; C505689:Tnfsf11 protein, mouse