Abe T, Murakami M, Sato T, Kajiki M, Ohno M, Kodaira R
Medical Science Laboratory, Asahi Chemical Industry Co., Ltd., Shizuoka-ken, Japan.
J Biol Chem. 1989 May 25;264(15):8941-5.
Human macrophage differentiation inducing factor (DIF) can induce differentiation of human myeloid leukemic cells into macrophage-like cells in vitro. A procedure is described for purification of DIF from serum-free human monocytic leukemia THP-1 cell-conditioned medium. The procedure included concentration of a conditioned medium by ultrafiltration, lentil lectin-Sepharose affinity chromatography, and fast protein liquid chromatography using Mono S and Mono Q. DIF-A of pI 9.0 and DIF-B of pI 8.8 were obtained after a final purification with a Mono Q column, and both DIF gave a single peak with a molecular weight of approximately 51,000 determined by gel chromatography. NH2-terminal amino acid analysis of DIF-A showed a noticeable homology with murine leukemia inhibitory factor. Human DIF-A was found to induce maturation of human and murine leukemic cells into both macrophage-like cells with nitro blue tetrazolium reducing activity and phagocytic cells, but was found to suppress proliferation of these leukemic cells.
人巨噬细胞分化诱导因子(DIF)可在体外诱导人髓系白血病细胞分化为巨噬细胞样细胞。本文描述了一种从无血清人单核细胞白血病THP - 1细胞条件培养基中纯化DIF的方法。该方法包括通过超滤浓缩条件培养基、扁豆凝集素 - 琼脂糖亲和层析以及使用Mono S和Mono Q的快速蛋白质液相色谱。在用Mono Q柱进行最终纯化后,获得了pI为9.0的DIF - A和pI为8.8的DIF - B,通过凝胶色谱法测定,两种DIF均给出了一个分子量约为51,000的单峰。对DIF - A的氨基末端氨基酸分析显示与小鼠白血病抑制因子有显著同源性。发现人DIF - A可诱导人和小鼠白血病细胞成熟为具有硝基蓝四氮唑还原活性的巨噬细胞样细胞和吞噬细胞,但发现它可抑制这些白血病细胞的增殖。