Hiraoka A, Nagasawa T, Ohta N, Sugimura A, Shimizu N, Ooi K, Shimizu S
Department of Internal Medicine, Osaka Dental University, Osaka, Japan.
Acta Haematol. 1998;100(4):174-80. doi: 10.1159/000040899.
Human stem cell growth factor (SCGF) produced by a myeloid cell line, KPB-M15, exhibits species-specific hematopoietic activities. However, KPB-M15-conditioned medium induced colony formation of mouse bone marrow cells. KPB-M15-derived colony-stimulating activity (CSA) was purified through Butyl-Toyopearl 650c and Cu2+ chelating-Sepharose 6B chromatography. TSK-G3000SW gel filtration of the purified preparation presented 3 distinct peaks around Vo, 150 kD and 85 kD. Gel fractions extracted from SDS-PAGE had macrophage colony-stimulating factor (M-CSF)-specific amino acid sequences. PCR, Northern hybridization and ELISA demonstrated that KPB-M15 cells secreted a significant amount of M-CSF and IL-6. Anti-M-CSF but not anti-IL-6 antibody abrogated CSA in KPB-M15-CM. IL-6 hardly synergized with M-CSF to enhance colony formation. Collectively, M-CSF is a sole CSA for murine hematopoietic progenitor cells in KPB-M15-CM. This is the first report of a human myeloid cell line, KPB-M15, constitutively producing M-CSF in addition to SCGF and IL-6. It can be useful in investigating the mechanism of production of M-CSF.
由髓系细胞系KPB - M15产生的人干细胞生长因子（SCGF）具有种属特异性造血活性。然而，KPB - M15条件培养基可诱导小鼠骨髓细胞形成集落。通过丁基 - 托普雷斯650c和Cu2 +螯合 - 琼脂糖6B层析纯化KPB - M15来源的集落刺激活性（CSA）。对纯化制剂进行TSK - G3000SW凝胶过滤，在空体积、150 kD和85 kD附近呈现3个不同的峰。从SDS - PAGE中提取的凝胶组分具有巨噬细胞集落刺激因子（M - CSF）特异性氨基酸序列。PCR、Northern杂交和ELISA表明KPB - M15细胞分泌大量的M - CSF和IL - 6。抗M - CSF抗体而非抗IL - 6抗体可消除KPB - M15条件培养基中的CSA。IL - 6几乎不与M - CSF协同增强集落形成。总体而言，M - CSF是KPB - M15条件培养基中小鼠造血祖细胞的唯一CSA。这是关于人髓系细胞系KPB - M15除了产生SCGF和IL - 6外还组成性产生M - CSF的首次报道。它可能有助于研究M - CSF的产生机制。
