ISOLATION AND CHARACTERIZATION OF A MACROPHAGE-DERIVED HEPARIN-BINDING GROWTH-FACTOR
ISOLATION AND CHARACTERIZATION OF A MACROPHAGE-DERIVED HEPARIN-BINDING GROWTH-FACTOR
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DOI：
10.1091/mbc.1.11.811
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发表时间：
1990-10-01
期刊：
影响因子：
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通讯作者：
KLAGSBRUN, M
中科院分区：
文献类型：
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作者：
BESNER, G;HIGASHIYAMA, S;KLAGSBRUN, M
将人单核细胞铺板于培养物中，并通过肝素-琼脂糖亲和层析分析这些细胞的条件培养基。通过刺激 BALB/c 3T3 细胞中的 DNA 合成来测试这些级分的生长因子活性。培养2 d后，检测到肝素结合生长因子（HBGF）活性的两个峰，一个用0.5 M NaCl洗脱，可以证明是血小板衍生生长因子（PDGF）样，另一个用1.0 M NaCl洗脱。培养 7-11 天后，当单核细胞明显分化为巨噬细胞时，条件培养基中 95% 以上的 HBGF 活性由 1.0 M NaCl 洗脱峰组成。这种活性被命名为巨噬细胞衍生的 HBGF (MD-HBGF)，被发现是一种分子量在 14-25 kDa 范围内的阳离子耐热多肽。使用蛋白质印迹和特异性中和抗血清进行的分析以及比较肝素亲和力分析表明，MD-HBGF 与已知由巨噬细胞产生的其他肝素结合 3T3 细胞生长因子不同，例如 PDGF（AB、AA 和 BB 形式）、酸性成纤维细胞生长因子和碱性成纤维细胞生长因子。除了刺激3T3细胞有丝分裂外，MD-HBGF还刺激血管平滑肌细胞的增殖，但不刺激血管内皮细胞的增殖。
Human mononuclear cells were plated in culture, and the conditioned media of these cells were analyzed by heparin-Sepharose affinity chromatography. The fractions were tested for growth factor activity as measured by the stimulation of DNA synthesis in BALB/c 3T3 cells. After 2 d in culture, two peaks of heparin-binding growth factor (HBGF) activity were detected, one eluting with 0.5 M NaCl, which could be shown to be platelet-derived growth factor (PDGF)-like, and the other eluting with 1.0 M NaCl. After 7-11 d in culture, when monocytes had clearly differentiated into macrophages, > 95% of the HBGF activity in conditioned medium consisted of the 1.0 M NaCl elution peak. This activity, which was designated macrophage-derived HBGF (MD-HBGF), was found to be a cationic heat-resistant polypeptide with a molecular weight in the range of 14-25 kDa. Analysis using Western blots and specific neutralizing antisera, as well as comparative heparin affinity analysis, indicated that MD-HBGF was not identical to other heparin-binding 3T3 cell growth factors known to be produced by macrophages, such as PDGF (AB, AA, and BB forms), acidic fibroblast growth factor, and basic fibroblast growth factor. In addition to stimulating mitogenesis in 3T3 cells, MD-HBGF also stimulated the proliferation of vascular smooth muscle cells, but did not stimulate the proliferation of vascular endothelial cells.