Characterization of osteoclast precursors in human blood
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 osteoproteg...
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| Veröffentlicht in: | British journal of haematology 2000-11, Vol.111 (2), p.501-512 |
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| creator | SHALHOUB, V ELLIOTT, G VAN, G SCULLY, S QI, M GRISANTI, M DUNSTAN, C BOYLE, W. J LACEY, D. L CHIU, L MANOUKIAN, R KELLEY, M HAWKINS, N DAVY, E SHIMAMOTO, G BECK, J KAUFMAN, S. A |
| description | 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. |
| doi_str_mv | 10.1046/j.1365-2141.2000.02379.x |
| format | Article |
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J ; LACEY, D. L ; CHIU, L ; MANOUKIAN, R ; KELLEY, M ; HAWKINS, N ; DAVY, E ; SHIMAMOTO, G ; BECK, J ; KAUFMAN, S. A</creator><creatorcontrib>SHALHOUB, V ; ELLIOTT, G ; VAN, G ; SCULLY, S ; QI, M ; GRISANTI, M ; DUNSTAN, C ; BOYLE, W. J ; LACEY, D. L ; CHIU, L ; MANOUKIAN, R ; KELLEY, M ; HAWKINS, N ; DAVY, E ; SHIMAMOTO, G ; BECK, J ; KAUFMAN, S. A</creatorcontrib><description>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.</description><identifier>ISSN: 0007-1048</identifier><identifier>EISSN: 1365-2141</identifier><identifier>DOI: 10.1046/j.1365-2141.2000.02379.x</identifier><identifier>PMID: 11122091</identifier><identifier>CODEN: BJHEAL</identifier><language>eng</language><publisher>Oxford: Blackwell</publisher><subject>B-Lymphocytes - metabolism ; Biological and medical sciences ; Carrier Proteins ; Cell Differentiation ; Cell differentiation, maturation, development, hematopoiesis ; Cell physiology ; Cells, Cultured ; Colony-Stimulating Factors - pharmacology ; Dose-Response Relationship, Drug ; Flow Cytometry ; Fundamental and applied biological sciences. Psychology ; Glycoproteins - metabolism ; Glycoproteins - pharmacology ; Hematology ; Humans ; Leukocytes, Mononuclear - metabolism ; Leukocytes, Mononuclear - ultrastructure ; Membrane Glycoproteins ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Molecular and cellular biology ; Monocytes - metabolism ; Osteoclasts - metabolism ; Osteoclasts - ultrastructure ; Osteoprotegerin ; Protein Binding ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, Cytoplasmic and Nuclear - metabolism ; Receptors, Tumor Necrosis Factor - metabolism</subject><ispartof>British journal of haematology, 2000-11, Vol.111 (2), p.501-512</ispartof><rights>2001 INIST-CNRS</rights><rights>Copyright Blackwell Scientific Publications Ltd. Nov 2000</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c432t-a6bb792575b08427a1430a45d0bd769c65cb20a84d034fd46ae1e1c13e277a8a3</citedby><cites>FETCH-LOGICAL-c432t-a6bb792575b08427a1430a45d0bd769c65cb20a84d034fd46ae1e1c13e277a8a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27915,27916</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=893215$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11122091$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>SHALHOUB, V</creatorcontrib><creatorcontrib>ELLIOTT, G</creatorcontrib><creatorcontrib>VAN, G</creatorcontrib><creatorcontrib>SCULLY, S</creatorcontrib><creatorcontrib>QI, M</creatorcontrib><creatorcontrib>GRISANTI, M</creatorcontrib><creatorcontrib>DUNSTAN, C</creatorcontrib><creatorcontrib>BOYLE, W. J</creatorcontrib><creatorcontrib>LACEY, D. L</creatorcontrib><creatorcontrib>CHIU, L</creatorcontrib><creatorcontrib>MANOUKIAN, R</creatorcontrib><creatorcontrib>KELLEY, M</creatorcontrib><creatorcontrib>HAWKINS, N</creatorcontrib><creatorcontrib>DAVY, E</creatorcontrib><creatorcontrib>SHIMAMOTO, G</creatorcontrib><creatorcontrib>BECK, J</creatorcontrib><creatorcontrib>KAUFMAN, S. A</creatorcontrib><title>Characterization of osteoclast precursors in human blood</title><title>British journal of haematology</title><addtitle>Br J Haematol</addtitle><description>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.</description><subject>B-Lymphocytes - metabolism</subject><subject>Biological and medical sciences</subject><subject>Carrier Proteins</subject><subject>Cell Differentiation</subject><subject>Cell differentiation, maturation, development, hematopoiesis</subject><subject>Cell physiology</subject><subject>Cells, Cultured</subject><subject>Colony-Stimulating Factors - pharmacology</subject><subject>Dose-Response Relationship, Drug</subject><subject>Flow Cytometry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glycoproteins - metabolism</subject><subject>Glycoproteins - pharmacology</subject><subject>Hematology</subject><subject>Humans</subject><subject>Leukocytes, Mononuclear - metabolism</subject><subject>Leukocytes, Mononuclear - ultrastructure</subject><subject>Membrane Glycoproteins</subject><subject>Microscopy, Confocal</subject><subject>Microscopy, Electron, Scanning</subject><subject>Molecular and cellular biology</subject><subject>Monocytes - metabolism</subject><subject>Osteoclasts - metabolism</subject><subject>Osteoclasts - ultrastructure</subject><subject>Osteoprotegerin</subject><subject>Protein Binding</subject><subject>RANK Ligand</subject><subject>Receptor Activator of Nuclear Factor-kappa B</subject><subject>Receptors, Cytoplasmic and Nuclear - metabolism</subject><subject>Receptors, Tumor Necrosis Factor - metabolism</subject><issn>0007-1048</issn><issn>1365-2141</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkE1LxDAURYMozjj6F6QouGvNS9KmXcrgFwy40XV4TVOmpW3GpIXRX2_rlBFcvcU99_I4hARAI6Aiua8j4EkcMhAQMUppRBmXWbQ_IctjcEqWYyLDsZAuyIX3NaXAaQznZAEAjNEMliRdb9Gh7o2rvrGvbBfYMrC-N1Y36Ptg54wenLfOB1UXbIcWuyBvrC0uyVmJjTdX812Rj6fH9_VLuHl7fl0_bEItOOtDTPJcZiyWcU5TwSSC4BRFXNC8kEmmk1jnjGIqCspFWYgEDRjQwA2TElPkK3J32N05-zkY36u28to0DXbGDl5JJlIhRTaCN__A2g6uG39TkI0IMMFHKD1A2lnvnSnVzlUtui8FVE1qVa0mg2oyqCa16let2o_V63l_yFtT_BVnlyNwOwPoNTalw05X_silGWcQ8x9Fp4BO</recordid><startdate>20001101</startdate><enddate>20001101</enddate><creator>SHALHOUB, V</creator><creator>ELLIOTT, G</creator><creator>VAN, G</creator><creator>SCULLY, S</creator><creator>QI, M</creator><creator>GRISANTI, M</creator><creator>DUNSTAN, C</creator><creator>BOYLE, W. 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Psychology</topic><topic>Glycoproteins - metabolism</topic><topic>Glycoproteins - pharmacology</topic><topic>Hematology</topic><topic>Humans</topic><topic>Leukocytes, Mononuclear - metabolism</topic><topic>Leukocytes, Mononuclear - ultrastructure</topic><topic>Membrane Glycoproteins</topic><topic>Microscopy, Confocal</topic><topic>Microscopy, Electron, Scanning</topic><topic>Molecular and cellular biology</topic><topic>Monocytes - metabolism</topic><topic>Osteoclasts - metabolism</topic><topic>Osteoclasts - ultrastructure</topic><topic>Osteoprotegerin</topic><topic>Protein Binding</topic><topic>RANK Ligand</topic><topic>Receptor Activator of Nuclear Factor-kappa B</topic><topic>Receptors, Cytoplasmic and Nuclear - metabolism</topic><topic>Receptors, Tumor Necrosis Factor - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>SHALHOUB, V</creatorcontrib><creatorcontrib>ELLIOTT, G</creatorcontrib><creatorcontrib>VAN, G</creatorcontrib><creatorcontrib>SCULLY, S</creatorcontrib><creatorcontrib>QI, M</creatorcontrib><creatorcontrib>GRISANTI, M</creatorcontrib><creatorcontrib>DUNSTAN, C</creatorcontrib><creatorcontrib>BOYLE, W. 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A</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>British journal of haematology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>SHALHOUB, V</au><au>ELLIOTT, G</au><au>VAN, G</au><au>SCULLY, S</au><au>QI, M</au><au>GRISANTI, M</au><au>DUNSTAN, C</au><au>BOYLE, W. J</au><au>LACEY, D. L</au><au>CHIU, L</au><au>MANOUKIAN, R</au><au>KELLEY, M</au><au>HAWKINS, N</au><au>DAVY, E</au><au>SHIMAMOTO, G</au><au>BECK, J</au><au>KAUFMAN, S. A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of osteoclast precursors in human blood</atitle><jtitle>British journal of haematology</jtitle><addtitle>Br J Haematol</addtitle><date>2000-11-01</date><risdate>2000</risdate><volume>111</volume><issue>2</issue><spage>501</spage><epage>512</epage><pages>501-512</pages><issn>0007-1048</issn><eissn>1365-2141</eissn><coden>BJHEAL</coden><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.</abstract><cop>Oxford</cop><pub>Blackwell</pub><pmid>11122091</pmid><doi>10.1046/j.1365-2141.2000.02379.x</doi><tpages>12</tpages></addata></record> |
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| subjects | B-Lymphocytes - metabolism Biological and medical sciences Carrier Proteins Cell Differentiation Cell differentiation, maturation, development, hematopoiesis Cell physiology Cells, Cultured Colony-Stimulating Factors - pharmacology Dose-Response Relationship, Drug Flow Cytometry Fundamental and applied biological sciences. Psychology Glycoproteins - metabolism Glycoproteins - pharmacology Hematology Humans Leukocytes, Mononuclear - metabolism Leukocytes, Mononuclear - ultrastructure Membrane Glycoproteins Microscopy, Confocal Microscopy, Electron, Scanning Molecular and cellular biology Monocytes - metabolism Osteoclasts - metabolism Osteoclasts - ultrastructure Osteoprotegerin Protein Binding RANK Ligand Receptor Activator of Nuclear Factor-kappa B Receptors, Cytoplasmic and Nuclear - metabolism Receptors, Tumor Necrosis Factor - metabolism |
| title | Characterization of osteoclast precursors in human blood |
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