Eosinophil chemotactic factor-1 (ECF-L): A novel osteoclast stimulating factor
Eosinophil chemotactic factor-1 (ECF-L): A novel osteoclast stimulating factor
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DOI：
10.1359/jbmr.2003.18.7.1332
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发表时间：
2003-07-01
影响因子：
6.2
通讯作者：
Roodman, GD
中科院分区：
文献类型：
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作者：
Oba, Y;Chung, HY;Roodman, GD
筛选富含OCL表达基因的cDNA文库鉴定ECF-L。ECF-L增强OCL形成而不增加RANKL水平。抗ECF-L抑制RANKL诱导的OCL形成。这些结果支持ECF-L在破骨细胞生成中的有效作用。引言：为了研究控制破骨细胞生成的分子机制，我们开发了一种永生化的破骨细胞（OCL）前体细胞系，该细胞系在没有基质细胞的情况下形成成熟的OCL，并将其用于形成纯OCL群体。使用聚合酶链反应（PCR）选择性cDNA消减法来鉴定在成熟OCL中与OCL前体相比高表达的基因，所述基因使用来自该细胞系的OCL和OCL前体。嗜酸性粒细胞趋化因子-L（ECF-L），以前描述的嗜酸性粒细胞的趋化因子，是鉴定的基因之一。在用10（-10）M 1，25（OH）（2）D-3处理的小鼠骨髓培养物中，用mECF-L cDNA或纯化的ECF-L Fc蛋白转染的293细胞的条件培养基以剂量依赖性方式增加OCL形成。与1，25（OH）（2）D-3诱导的OCL相比，ECF-L条件培养液诱导的OCL在每个牙本质切片上形成的小窝数量和吸收面积增加（p < 0.01）。与正义S-寡核苷酸对照相比，在仅用10（-9）M 1，25（OH）（2）D-3处理的小鼠骨髓培养物中，向mECF-L中加入反义S-寡核苷酸可抑制OCL形成。时程研究表明，ECF-L在OCL形成的后期阶段起作用，趋化性测定表明mECF-L增加OCL前体的迁移。原位杂交检测到单核细胞和多核细胞中的mECF-L mRNA。有趣的是，ECF-L的中和抗体阻断了RANKL或10（-9）M 1，25（OH）（2）D-3诱导的小鼠骨髓培养物中OCL的形成，尽管ECF-L并不诱导RANKL的表达。结论：这些数据表明ECF-L是一种以前未知的因子，是OCL形成的有效介质，其在OCL形成的后期阶段起作用并增强RANKL的作用。
Screening a cDNA library enriched for genes expressed in OCLs identified 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)(2)D-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)(2)D-3 (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)(2)D-3 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)(2)D-3-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.