Brown W C, Grab D J
J Immunol. 1985 Nov;135(5):3184-90.
Bovine peripheral blood lymphocytes stimulated with the T cell mitogen concanavalin A (Con A) secrete a lymphokine with biological properties similar to T cell growth factor (TCGF) or interleukin 2 (IL 2) from other species. The material supports proliferation of Con A-derived T cell blasts, limiting dilution cloning of T cell blasts, and continuous growth of T cell clones for over 6 mo in vitro. A quantitative microassay with the use of TCGF-dependent, Con A-unresponsive cloned T cells was used to determine the biological activity during purification of IL 2. A single peak of activity with an apparent m.w. of 25,000 to 28,000 was recovered after gel filtration. This material eluted from DEAE-Sephacryl between 135 and 165 mM NaCl. After isoelectric focusing, high pressure liquid chromatography, and gel electrophoresis under reducing conditions, peak IL 2 activity was associated with proteins having m.w. of 20,000 and 23,000.
用T细胞促有丝分裂原刀豆球蛋白A(Con A)刺激牛外周血淋巴细胞,可分泌一种淋巴因子,其生物学特性与其他物种的T细胞生长因子(TCGF)或白细胞介素2(IL 2)相似。该物质支持Con A诱导的T细胞母细胞增殖、T细胞母细胞的有限稀释克隆以及T细胞克隆在体外持续生长超过6个月。在IL 2纯化过程中,使用依赖TCGF、对Con A无反应的克隆T细胞进行定量微量测定,以确定其生物活性。凝胶过滤后,得到一个表观分子量为25,000至28,000的单一活性峰。该物质在135至165 mM NaCl浓度下从DEAE-葡聚糖凝胶上洗脱下来。在等电聚焦、高压液相色谱和还原条件下的凝胶电泳后,IL 2活性峰与分子量为20,000和23,000的蛋白质相关。