Hasebe A, Shibata K, Domon H, Dong L, Watanabe T
Department of Oral Bacteriology, Hokkaido University School of Dentistry, Sapporo, Japan.
Microbiol Immunol. 1999;43(11):1003-8. doi: 10.1111/j.1348-0421.1999.tb01229.x.
The active entity responsible for inducing interleukin-6 production by human gingival fibroblasts was partially purified by ion-exchange chromatography from the water-soluble fraction of Mycoplasma salivarium cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the final preparation revealed one densely stained band with a molecular weight of 20.6 kilodaltons and two faint bands with molecular weights of 40.5 and 82.5 kilodaltons. The specific activity of the final preparation was 34-fold higher than that of the starting water-soluble fraction. The interleukin-6-inducing activity was destroyed by proteinase K and reduced 70% by lipoprotein lipase and heat treatment, but was not affected by deoxyribonuclease I or endoglucosidase D. The final preparation induced small amounts of tumor necrosis factor-alpha and interleukin-lbeta in a myelomonocytic cell line, THP-1 cells, but did not induce interleukin-6. The ability of Escherichia coli lipopolysaccharide to stimulate human gingival fibroblasts to release interleukin-6 was dependent upon the presence of serum in the assay medium, but that of the final preparation from M. salivarium was not. Thus, we partially purified the protein(s) from M. salivarium which were capable of stimulating human gingival fibroblasts to release interleukin-6 by a mechanism different from that of E. coli lipopolysaccharide.
通过离子交换色谱法从唾液支原体细胞的水溶性部分中部分纯化了负责诱导人牙龈成纤维细胞产生白细胞介素-6的活性物质。最终制剂的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示出一条分子量为20.6千道尔顿的深染带和两条分子量分别为40.5和82.5千道尔顿的淡染带。最终制剂的比活性比起始水溶性部分高34倍。白细胞介素-6诱导活性被蛋白酶K破坏，经脂蛋白脂肪酶和热处理后降低70%，但不受脱氧核糖核酸酶I或内切葡糖苷酶D影响。最终制剂在髓单核细胞系THP-1细胞中诱导产生少量肿瘤坏死因子-α和白细胞介素-1β，但不诱导白细胞介素-6。大肠杆菌脂多糖刺激人牙龈成纤维细胞释放白细胞介素-6的能力取决于测定培养基中血清的存在，而唾液支原体的最终制剂则不然。因此，我们从唾液支原体中部分纯化了能够通过不同于大肠杆菌脂多糖的机制刺激人牙龈成纤维细胞释放白细胞介素-6的蛋白质。
