Higuchi M, Sugimoto M, Kobayashi Y, Osawa T
Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, University of Tokyo.
Microbiol Immunol. 1987;31(5):481-90. doi: 10.1111/j.1348-0421.1987.tb03110.x.
Two different factors (MAF-C I and MAF-C II) were obtained by anion exchange chromatography of the culture supernatant of a human T-cell hybridoma, H3-E9-6, which produces macrophage-activating factors for cytotoxicity (MAF-C). These 2 factors induced the cytotoxicity of monocytes synergistically as a priming signal (MAF-C I) and a triggering signal (MAF-C II), respectively. On gel filtration on a column of Superose 12, MAF-C II was eluted mainly at the void volume, whereas MAF-C I was eluted in the fractions corresponding to approximate molecular weights of 30-300 K. On the other hand, gel filtration in the presence of sodium deoxycholate revealed that MAF-C II has an approximate molecular weight of 40,000, but MAF-C I was unstable under these conditions. When the activity for mouse macrophages (MAF-Cm activity) was tested, the MAF-C II fraction showed high MAF-Cm activity in the presence of murine recombinant interferon gamma (rIFN-gamma), but the MAF-C I fraction did not show MAF-Cm activity even in the presence of lipopolysaccharide (LPS). These results suggest that MAF-C I (priming lymphokine) has species specificity but MAF-C II (triggering lymphokine) does not.
通过对人T细胞杂交瘤H3-E9-6的培养上清液进行阴离子交换色谱分析,获得了两种不同的因子(MAF-C I和MAF-C II),该杂交瘤可产生具有细胞毒性的巨噬细胞激活因子(MAF-C)。这两种因子分别作为启动信号(MAF-C I)和触发信号(MAF-C II)协同诱导单核细胞的细胞毒性。在Superose 12柱上进行凝胶过滤时,MAF-C II主要在空体积处洗脱,而MAF-C I在对应于约30-300K分子量的级分中洗脱。另一方面,在脱氧胆酸钠存在下进行凝胶过滤显示,MAF-C II的分子量约为40,000,但MAF-C I在这些条件下不稳定。当测试对小鼠巨噬细胞的活性(MAF-Cm活性)时,MAF-C II级分在存在鼠重组干扰素γ(rIFN-γ)的情况下显示出高MAF-Cm活性,但MAF-C I级分即使在存在脂多糖(LPS)的情况下也未显示出MAF-Cm活性。这些结果表明,MAF-C I(启动淋巴细胞因子)具有种属特异性,而MAF-C II(触发淋巴细胞因子)则没有。