Minta J O
Department of Pathology, University of Toronto, Ontario, Canada.
J Immunol. 1988 Sep 1;141(5):1636-41.
The human pre-monocytic cell line U-937 was shown to synthesize and to secrete increasing amounts of factor B in short term cultures in serum-free medium containing BSA. The kinetics of factor B production were higher on day 2 than on days 1 and 3. The production of factor B was reversibly inhibited by cycloheximide, indicating de novo synthesis. Metabolic labeling with [35S]-methionine and SDS-PAGE analysis revealed that both intracellular and secreted factor B were single-chain proteins with similar m.w. (90,000), which co-migrated with purified plasma factor B. Incubation of U-937 cells with the immunostimulants PMA, LPS, IFN-gamma, and IL-1 resulted in a dose-dependent augmentation of factor B production. A 24-h exposure to IL-1 was shown to be required for maximal stimulation. A combination of suboptimal doses of LPS and IFN-gamma was shown to exert a synergistic effect on factor B production. The U-937 cell line is thus a valuable model for the study of the regulation of the factor B gene expression.
人单核细胞前体细胞系U - 937在含牛血清白蛋白的无血清培养基中短期培养时,可合成并分泌越来越多的B因子。B因子产生的动力学在第2天高于第1天和第3天。环己酰亚胺可可逆性抑制B因子的产生,表明其为从头合成。用[35S] - 甲硫氨酸进行代谢标记及SDS - PAGE分析显示,细胞内和分泌的B因子均为单链蛋白,分子量相似(90,000),与纯化的血浆B因子迁移率相同。用免疫刺激剂佛波酯(PMA)、脂多糖(LPS)、γ干扰素(IFN - γ)和白细胞介素 - 1(IL - 1)孵育U - 937细胞,可导致B因子产生呈剂量依赖性增加。显示需要24小时暴露于IL - 1才能达到最大刺激效果。亚最佳剂量的LPS和IFN - γ组合对B因子产生具有协同作用。因此,U - 937细胞系是研究B因子基因表达调控的有价值模型。