Characterization of osteoclast precursors in human blood
Characterization of osteoclast precursors in human blood
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DOI：
10.1046/j.1365-2141.2000.02379.x
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发表时间：
2000-11-01
影响因子：
6.5
通讯作者：
Lacey, DL
中科院分区：
文献类型：
--
作者：
Shalhoub, V;Elliott, G;Lacey, DL
破骨细胞前体（OCPs）在外周血（PB）的单核部分中循环，但其丰度和表面特征尚不清楚。先前的研究表明，小鼠骨髓中马槟榔处理的OCP上的NF-κ B受体激活剂（RANK）与骨保护素配体（OPGL/TRANCE/RANKL/ODF）相互作用，启动破骨细胞分化。因此，我们使用荧光形式的人OPGL（Hu-OPGL-F），通过荧光激活细胞分选分析，在未处理的外周血单核细胞（PBMC）中鉴定可能表达RANK的OCP。单核细胞[CD 14-藻红蛋白（PE）抗体（Ab）阳性（+）细胞，10-15%的PBMC]均（98-100%）与Hu-OPGL-F共标记（n > 18）。T淋巴细胞（CD 3-PE Ab（+）细胞，占PBMC的66%）不结合Hu-OPGL-F;然而，B细胞（CD 19-PE Ab（+）细胞，占PBMC的9%）也为Hu-OPGL-F阳性。所有Hu-OPGL-F+单核细胞还与CD 33、CD 61、CD 11b、CD 38、CD 45和CD 54 Ab共标记，但不与CD 34或CD 56 Ab共标记。Hu-OPGL-F结合呈剂量依赖性，并与过量Hu-OPGL竞争。当Hu-OPGL-F+、CD 14-PE Ab（+）、CD 33-PE Ab（+）、Hu-OPGL-F+/CD 14-PE Ab（+）或Hu-OPGL-F+/CD 33-PE Ab（+）细胞与OPGL（20 ng/ml）和集落刺激因子（CSF）-1（25 ng/ml）一起培养时，OC样细胞很容易发育。因此，所有新鲜分离的单核细胞均表现出可置换的Hu-OPGL-F结合，表明PB中OCP上存在RANK;此外，在完全不存在其他细胞类型的情况下，在含OPGL和CSF-1的培养基中，纯化的PB单核细胞群中的OCP形成破骨细胞样细胞。
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.