Schneiderman J, Knoller S, Adar R, Savion N
Department of General and Vascular Surgery, Sheba Medical Center, Tel-Hashomer, Israel.
J Vasc Surg. 1991 Jul;14(1):103-10.
A specific inhibitory factor was isolated from human serum of a patient who manifested impaired prosthetic graft tissue incorporation. The latter reaction was hypothesized to be related to inhibiting fibroblast proliferation by a specific humoral factor. The crude inhibitory serum was tested against a pool of normal human sera with different cell types in culture. Proliferation of human skin fibroblast and human smooth muscle cell cultures incubated in the presence of 50% inhibitory serum were inhibited up to 58% and 37%, respectively. Proliferation of bovine capillary endothelial cell cultures was stimulated under similar conditions. Isolation and purification of the inhibitory factor from crude serum were initiated by ammonium sulfate precipitation. The pellet was further fractionated by sepharose 6B gel filtration. The inhibitory activity was eluted from the column in a relatively purified form as indicated by gel electrophoresis of the inhibitory fractions, which demonstrated a specific band corresponding to the inhibitory protein, with an apparent relative molecular mass of 230 kDa. The inhibitory factor showed a high affinity to concanavalin A, indicating its nature as a glycoprotein not associated with albumin or immunoglobulin fractions of the serum.
从一名人工血管移植组织整合受损患者的人血清中分离出一种特异性抑制因子。推测后一种反应与一种特异性体液因子抑制成纤维细胞增殖有关。将粗制抑制血清与培养中不同细胞类型的正常人血清池进行测试。在50%抑制血清存在下培养的人皮肤成纤维细胞和人平滑肌细胞培养物的增殖分别被抑制高达58%和37%。在类似条件下，牛毛细血管内皮细胞培养物的增殖受到刺激。通过硫酸铵沉淀开始从粗血清中分离和纯化抑制因子。沉淀物通过琼脂糖6B凝胶过滤进一步分级分离。如抑制级分的凝胶电泳所示，抑制活性以相对纯化的形式从柱中洗脱出来，其显示出对应于抑制蛋白的特异性条带，表观相对分子质量为230 kDa。该抑制因子对伴刀豆球蛋白A具有高亲和力，表明其性质为不与血清的白蛋白或免疫球蛋白部分相关的糖蛋白。
