人绒毛膜促性腺激素的纯化及其对小鼠脾细胞产生CSFs的诱导作用

HCG 粗品(10Iu/mg)经乙醇抽提,Sephadex G-100和 SE-Sephadex C-50凝胶过滤后,纯化了800倍,免疫活性达8370Iu/mg.经 SDS-PAGE 鉴定 HCG 的两亚基分子量分别为17000和32000.用纯化的 HCG 制备具有较高滴度的抗 HCG 抗血清.通过软琼脂培养法观察了 HCG 诱导小鼠脾细胞产生 CSFs 的影响,结果表明 HCG 浓度为400Iu/mg时,诱导效应最大。时间依赖曲线以72小时的 CSFs 活力最高.HCG 的诱导作用可以被抗HCG 抗血清完全中和。

PURIFICATION OF HUMAN CHORIONIC GONADOTROPIN AND INDUCTION OF COLONY STIMULATING FACTOR IN THE MICE''''SPLEEN CELL

The purification of crude HCG (10Iu/mg) was involved with ethanol fractionation and gel filtration on sephadex C-100 and SE-sephadex C-50.The prepara- tion was purified 800-fold and was assayed immunologically at 8730Iu/mg.The molecu- lar weight of HCG subunits of 17,000 and 30,000 was determined by SDS-PAGE.In addi- tion we succeeded in producing anti-HCG serum which had high titre assayed by immunodifussion. HCG can stimulate the spleen cell (B lymphocyte) of mice to secrete colony stimulating factor (CSFs) with dose dependent and time depedent fashion.The dose-dependent re- sponse curve shows that the amount of CSFs is the highest when HCG dose is 200-400Iu/ml.The time-dependent response curve shows that the most effective activity of CSFs is after 72 hours.The induction of CSFs by HCG can be neutralized by anti-HCG serum in vitro formation of colonies from spleen cells in soft agar culture.