Gaffney E V, Dell'Aquila M L, Lingenfelter S E, Huffnagle G B, Wiest D L
J Leukoc Biol. 1986 Apr;39(4):409-21. doi: 10.1002/jlb.39.4.409.
The human monocytic leukemia cell line, THP-1, acquires macrophage-like characteristics following exposure to mezerein. Serum-free medium conditioned by mezerein-activated cells was observed to contain colony-stimulating factor (CSF) activity in assays with murine bone marrow cultures. Isoelectrofocusing revealed that CSF activity displayed charge heterogeneity and migrated in a pl range of 4.4-5.3. Treatment with neuraminidase did not affect biological activity but did reduce charge heterogeneity. Reisofocusing of neuraminidase-treated CSF revealed a peak of activity at pl 4.9. The active component was shown to be an acidic sialoglycoprotein, resistant to proteolytic cleavage but completely inactivated by 2-mercaptoethanol. This CSF has been purified from THP-1-conditioned serum-free medium by preparative isoelectrofocusing, gel filtration through Sephacryl S-200, ion exchange chromatography on DEAE-Sephacel, neuraminidase treatment, and tris-glycinate polyacrylamide gel electrophoresis (PAGE). Elution from SDS-PAGE revealed a single peak of activity corresponding to an apparent molecular weight of 70,000 daltons. Preliminary characterization of the bone marrow cells in colonies showed that THP-1 cells produced macrophage-specific CSF.
人单核细胞白血病细胞系THP-1在暴露于狼毒素后会获得巨噬细胞样特征。在用小鼠骨髓培养物进行的试验中，观察到经狼毒素激活的细胞所分泌的无血清培养基含有集落刺激因子（CSF）活性。等电聚焦显示，CSF活性呈现电荷异质性，在pH值范围4.4 - 5.3内迁移。用神经氨酸酶处理不影响生物活性，但会降低电荷异质性。对经神经氨酸酶处理的CSF进行再等电聚焦显示，在pH 4.9处有一个活性峰。活性成分被证明是一种酸性唾液糖蛋白，对蛋白水解裂解具有抗性，但被2-巯基乙醇完全灭活。这种CSF已通过制备性等电聚焦、经Sephacryl S-200凝胶过滤、在DEAE-Sephacel上进行离子交换色谱、神经氨酸酶处理以及Tris-甘氨酸聚丙烯酰胺凝胶电泳（PAGE）从THP-1条件性无血清培养基中纯化出来。从SDS-PAGE洗脱显示出一个单一的活性峰，对应于表观分子量为70,000道尔顿。对集落中的骨髓细胞进行的初步表征表明，THP-1细胞产生巨噬细胞特异性CSF。
