Establishment of Neutralizing Rat Monoclonal Antibodies for Fibroblast Growth Factor-2
Establishment of Neutralizing Rat Monoclonal Antibodies for Fibroblast Growth Factor-2
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DOI：
10.1089/mab.2013.0085
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发表时间：
2014-08-01
影响因子：
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通讯作者：
Ohkawa, Yasuyuki
中科院分区：
文献类型：
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作者：
Tanaka, Masako;Yamaguchi, Maki;Ohkawa, Yasuyuki
成纤维细胞生长因子-2（FGF-2）在内皮细胞存活、增殖和血管生成中起关键作用，并通过与硫酸乙酰肝素蛋白聚糖结合而定位于细胞膜上。在这里，我们建立了一个中和单克隆抗体，1B 9 B 9，对FGF-2，使用大鼠髂内淋巴结的方法。1B 9 B 9阻断FGF-2与其受体的结合，抑制FGF-2诱导的内皮细胞增殖和相应的下游信号传导。用1B 9 B 9处理人脐静脉内皮细胞可降低Akt和MAPK的基础磷酸化水平。此外，继续用1B 9 B 9处理通过细胞凋亡诱导细胞死亡。与FGF-2敲低相比，1B 9 B 9显著降低细胞存活。此外，FGF-2 siRNA和1B 9 B 9的组合显示出协同效应。数据表明，通过大鼠髂淋巴结方法建立的1B 9 B 9是一种完全相容的中和抗体。
Fibroblast growth factor-2 (FGF-2) plays a critical role in endothelial survival, proliferation, and angiogenesis and is localized on the cell membrane by binding to heparan sulfate proteoglycans. Here we established a neutralizing monoclonal antibody, 1B9B9, against FGF-2 using the rat medial iliac lymph node method. 1B9B9 blocked the binding of FGF-2 to its receptor, inhibiting FGF-2-induced proliferation and corresponding downstream signaling in endothelial cells. Treatment of human umbilical vein endothelial cells with 1B9B9 reduced the basal phosphorylation levels of Akt and MAPK. Furthermore, continued treatment with 1B9B9 induced cell death by apoptosis. Compared with FGF-2 knockdown, 1B9B9 significantly reduced cell survival. In addition, the combination of FGF-2 siRNA and 1B9B9 showed a synergistic effect. The data indicate that 1B9B9 established by the rat iliac lymph node method is a fully compatible neutralizing antibody.