Eosinophil Chemotactic Factor-L (ECF-L): A Novel Osteoclast Stimulating Factor

Yasuo Oba, Ho Yeon Chung, Sun Jin Choi, G. David Roodman

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Screening a cDNA library enriched for genes expressed in OCLs identitied ECF-L. ECF-L enhanced OCL formation without increasing RANKL levels. Anti-ECF-L inhibited RANKL-induced OCL formation. These results support a potent role of ECF-L in osteoclastogenesis. Introduction: To investigate the molecular mechanisms that control osteoclastogenesis, we developed an immortalized osteoclast (OCL) precursor cell line that forms mature OCLs in the absence of stromal cells and used it to form pure populations of OCLs. Materials and Methods: Polymerase chain reaction (PCR) selective cDNA subtraction was used to identify genes that are highly expressed in mature OCLs compared with OCL precursors employing OCL and OCL precursors derived from this cell line. Results: Eosinophil chemotactic factor-L (ECF-L), a previously described chemotactic factor for eosinophils, was one of the genes identified. Conditioned media from 293 cells transfected with mECF-L cDNA, or purified ECF-L Fc protein, increased OCL formation in a dose-dependent manner in mouse bone marrow cultures treated with 10-10 M 1,25(OH)2D 3. OCLs derived from marrow cultures treated with ECF-L conditioned media formed increased pit numbers and resorption area per dentin slice compared with OCLs induced by 1,25(OH)2D3 (p <0.01). Addition of an antisense S-oligonucleotide to mECF-L inhibited OCL formation in murine bone marrow cultures treated only with 10-9 M 1,25(OH) 2D3 compared with the sense S-oligonucleotide control. Time course studies demonstrated that ECF-L acted at the later stages of OCL formation, and chemotactic assays showed that mECF-L increased migration of OCL precursors. mECF-L mRNA was detectable in mononuclear and multinucleated cells by in situ hybridization. Interestingly, a neutralizing antibody to ECF-L blocked RANKL or 10-9 M 1,25(OH)2D3-induced OCL formation in mouse bone marrow cultures, although ECF-L did not induce RANKL expression. Conclusions: These data show ECF-L is a previously unknown factor that is a potent mediator of OCL formation, which acts at the later stages of OCL formation and enhances the effects of RANKL.

Original languageEnglish (US)
Pages (from-to)1332-1341
Number of pages10
JournalJournal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
Volume18
Issue number7
DOIs
StatePublished - Jul 1 2003
Externally publishedYes

Fingerprint

Eosinophil Chemotactic Factors
Osteoclasts
Bone Marrow
Conditioned Culture Medium
Osteogenesis
osteoclast stimulating factor
Complementary DNA
Genes
Cell Line
Antisense Oligonucleotides

Keywords

  • Eosinophil chemotactic factor-L
  • Fusion
  • Osteoclasts
  • RANKL

ASJC Scopus subject areas

  • Surgery

Cite this

Eosinophil Chemotactic Factor-L (ECF-L) : A Novel Osteoclast Stimulating Factor. / Oba, Yasuo; Chung, Ho Yeon; Choi, Sun Jin; Roodman, G. David.

In: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 18, No. 7, 01.07.2003, p. 1332-1341.

Research output: Contribution to journalArticle

@article{19260caffc4a43afaa330a001ba78e67,
title = "Eosinophil Chemotactic Factor-L (ECF-L): A Novel Osteoclast Stimulating Factor",
abstract = "Screening a cDNA library enriched for genes expressed in OCLs identitied ECF-L. ECF-L enhanced OCL formation without increasing RANKL levels. Anti-ECF-L inhibited RANKL-induced OCL formation. These results support a potent role of ECF-L in osteoclastogenesis. Introduction: To investigate the molecular mechanisms that control osteoclastogenesis, we developed an immortalized osteoclast (OCL) precursor cell line that forms mature OCLs in the absence of stromal cells and used it to form pure populations of OCLs. Materials and Methods: Polymerase chain reaction (PCR) selective cDNA subtraction was used to identify genes that are highly expressed in mature OCLs compared with OCL precursors employing OCL and OCL precursors derived from this cell line. Results: Eosinophil chemotactic factor-L (ECF-L), a previously described chemotactic factor for eosinophils, was one of the genes identified. Conditioned media from 293 cells transfected with mECF-L cDNA, or purified ECF-L Fc protein, increased OCL formation in a dose-dependent manner in mouse bone marrow cultures treated with 10-10 M 1,25(OH)2D 3. OCLs derived from marrow cultures treated with ECF-L conditioned media formed increased pit numbers and resorption area per dentin slice compared with OCLs induced by 1,25(OH)2D3 (p <0.01). Addition of an antisense S-oligonucleotide to mECF-L inhibited OCL formation in murine bone marrow cultures treated only with 10-9 M 1,25(OH) 2D3 compared with the sense S-oligonucleotide control. Time course studies demonstrated that ECF-L acted at the later stages of OCL formation, and chemotactic assays showed that mECF-L increased migration of OCL precursors. mECF-L mRNA was detectable in mononuclear and multinucleated cells by in situ hybridization. Interestingly, a neutralizing antibody to ECF-L blocked RANKL or 10-9 M 1,25(OH)2D3-induced OCL formation in mouse bone marrow cultures, although ECF-L did not induce RANKL expression. Conclusions: These data show ECF-L is a previously unknown factor that is a potent mediator of OCL formation, which acts at the later stages of OCL formation and enhances the effects of RANKL.",
keywords = "Eosinophil chemotactic factor-L, Fusion, Osteoclasts, RANKL",
author = "Yasuo Oba and Chung, {Ho Yeon} and Choi, {Sun Jin} and Roodman, {G. David}",
year = "2003",
month = "7",
day = "1",
doi = "10.1359/jbmr.2003.18.7.1332",
language = "English (US)",
volume = "18",
pages = "1332--1341",
journal = "Journal of Bone and Mineral Research",
issn = "0884-0431",
publisher = "Wiley-Blackwell",
number = "7",

}

TY - JOUR

T1 - Eosinophil Chemotactic Factor-L (ECF-L)

T2 - A Novel Osteoclast Stimulating Factor

AU - Oba, Yasuo

AU - Chung, Ho Yeon

AU - Choi, Sun Jin

AU - Roodman, G. David

PY - 2003/7/1

Y1 - 2003/7/1

N2 - Screening a cDNA library enriched for genes expressed in OCLs identitied ECF-L. ECF-L enhanced OCL formation without increasing RANKL levels. Anti-ECF-L inhibited RANKL-induced OCL formation. These results support a potent role of ECF-L in osteoclastogenesis. Introduction: To investigate the molecular mechanisms that control osteoclastogenesis, we developed an immortalized osteoclast (OCL) precursor cell line that forms mature OCLs in the absence of stromal cells and used it to form pure populations of OCLs. Materials and Methods: Polymerase chain reaction (PCR) selective cDNA subtraction was used to identify genes that are highly expressed in mature OCLs compared with OCL precursors employing OCL and OCL precursors derived from this cell line. Results: Eosinophil chemotactic factor-L (ECF-L), a previously described chemotactic factor for eosinophils, was one of the genes identified. Conditioned media from 293 cells transfected with mECF-L cDNA, or purified ECF-L Fc protein, increased OCL formation in a dose-dependent manner in mouse bone marrow cultures treated with 10-10 M 1,25(OH)2D 3. OCLs derived from marrow cultures treated with ECF-L conditioned media formed increased pit numbers and resorption area per dentin slice compared with OCLs induced by 1,25(OH)2D3 (p <0.01). Addition of an antisense S-oligonucleotide to mECF-L inhibited OCL formation in murine bone marrow cultures treated only with 10-9 M 1,25(OH) 2D3 compared with the sense S-oligonucleotide control. Time course studies demonstrated that ECF-L acted at the later stages of OCL formation, and chemotactic assays showed that mECF-L increased migration of OCL precursors. mECF-L mRNA was detectable in mononuclear and multinucleated cells by in situ hybridization. Interestingly, a neutralizing antibody to ECF-L blocked RANKL or 10-9 M 1,25(OH)2D3-induced OCL formation in mouse bone marrow cultures, although ECF-L did not induce RANKL expression. Conclusions: These data show ECF-L is a previously unknown factor that is a potent mediator of OCL formation, which acts at the later stages of OCL formation and enhances the effects of RANKL.

AB - Screening a cDNA library enriched for genes expressed in OCLs identitied ECF-L. ECF-L enhanced OCL formation without increasing RANKL levels. Anti-ECF-L inhibited RANKL-induced OCL formation. These results support a potent role of ECF-L in osteoclastogenesis. Introduction: To investigate the molecular mechanisms that control osteoclastogenesis, we developed an immortalized osteoclast (OCL) precursor cell line that forms mature OCLs in the absence of stromal cells and used it to form pure populations of OCLs. Materials and Methods: Polymerase chain reaction (PCR) selective cDNA subtraction was used to identify genes that are highly expressed in mature OCLs compared with OCL precursors employing OCL and OCL precursors derived from this cell line. Results: Eosinophil chemotactic factor-L (ECF-L), a previously described chemotactic factor for eosinophils, was one of the genes identified. Conditioned media from 293 cells transfected with mECF-L cDNA, or purified ECF-L Fc protein, increased OCL formation in a dose-dependent manner in mouse bone marrow cultures treated with 10-10 M 1,25(OH)2D 3. OCLs derived from marrow cultures treated with ECF-L conditioned media formed increased pit numbers and resorption area per dentin slice compared with OCLs induced by 1,25(OH)2D3 (p <0.01). Addition of an antisense S-oligonucleotide to mECF-L inhibited OCL formation in murine bone marrow cultures treated only with 10-9 M 1,25(OH) 2D3 compared with the sense S-oligonucleotide control. Time course studies demonstrated that ECF-L acted at the later stages of OCL formation, and chemotactic assays showed that mECF-L increased migration of OCL precursors. mECF-L mRNA was detectable in mononuclear and multinucleated cells by in situ hybridization. Interestingly, a neutralizing antibody to ECF-L blocked RANKL or 10-9 M 1,25(OH)2D3-induced OCL formation in mouse bone marrow cultures, although ECF-L did not induce RANKL expression. Conclusions: These data show ECF-L is a previously unknown factor that is a potent mediator of OCL formation, which acts at the later stages of OCL formation and enhances the effects of RANKL.

KW - Eosinophil chemotactic factor-L

KW - Fusion

KW - Osteoclasts

KW - RANKL

UR - http://www.scopus.com/inward/record.url?scp=0142091219&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0142091219&partnerID=8YFLogxK

U2 - 10.1359/jbmr.2003.18.7.1332

DO - 10.1359/jbmr.2003.18.7.1332

M3 - Article

C2 - 12854845

AN - SCOPUS:0142091219

VL - 18

SP - 1332

EP - 1341

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 7

ER -