Seckinger P, Vey E, Turcatti G, Wingfield P, Dayer J M
Division of Immunology and Allergy (Hans Wilsdorf Laboratory), Department of Medicine, Hôpital Cantonal Universitaire, Geneva, Switzerland.
Eur J Immunol. 1990 May;20(5):1167-74. doi: 10.1002/eji.1830200533.
The urine of some febrile patients has been shown to contain a tumor necrosis factor-alpha-inhibiting activity (TNF-alpha INH) when tested in a cytotoxicity assay using the TNF-susceptible cell line L-929. The inhibitor was purified to homogeneity using a simple three-step procedure which included a TNF-alpha affinity column, cation exchange and reverse-phase chromatography. The NH2-terminal amino acid sequence of the inhibitor showed no sequence similarity with proteins in the data bases used. Using gel filtration, it was shown that TNF-alpha and the inhibitor form a stable complex which eluted with a molecular weight of about 75,000. This value corresponds to the sum of the inhibitor (approximately 30,000) and TNF-alpha (approximately 45,000-50,000) molecular weight. The TNF-alpha INH blocked prostaglandin E2 production by dermal fibroblasts in a dose-dependent manner, providing evidence for antiinflammatory activity. TNF-alpha INH also blocked class I antigen expression in a dose-dependent manner as measured using the human Colo 205 tumor cell line. Furthermore, TNF-alpha INH affected TNF-alpha synergism with IFN-gamma-induced HLA-DR antigen expression but had no effect on IFN-gamma activity. The data presented demonstrate that TNF-alpha bioactivity can be regulated at the protein level.
当使用对肿瘤坏死因子（TNF）敏感的L-929细胞系进行细胞毒性试验时，一些发热患者的尿液显示含有肿瘤坏死因子α抑制活性（TNF-α INH）。通过一个简单的三步程序将该抑制剂纯化至同质，该程序包括TNF-α亲和柱、阳离子交换和反相色谱。该抑制剂的NH2末端氨基酸序列与所使用数据库中的蛋白质没有序列相似性。使用凝胶过滤法表明，TNF-α与该抑制剂形成了一个稳定的复合物，其洗脱分子量约为75,000。该值对应于抑制剂（约30,000）和TNF-α（约45,000 - 50,000）分子量之和。TNF-α INH以剂量依赖的方式阻断了真皮成纤维细胞中前列腺素E2的产生，为其抗炎活性提供了证据。TNF-α INH还以剂量依赖的方式阻断了使用人结肠癌细胞系Colo 205测量的I类抗原表达。此外，TNF-α INH影响了TNF-α与干扰素γ诱导的HLA-DR抗原表达的协同作用，但对干扰素γ的活性没有影响。所呈现的数据表明，TNF-α的生物活性可以在蛋白质水平上受到调节。
