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本文介绍了人类精浆免疫抑制因子分离及免疫活性测定的全过程。通过低温高速离心,SephadexG-100凝胶过滤,DEAE离子交换层析等步骤从人精浆中分离出两种免疫抑制因子——DF_1和DF_2。DF_2达到聚丙烯酰胺凝胶电泳纯,呈单一蛋白带,DF_1呈二条蛋白带。二者对PHA诱导的人外周血淋巴细胞转化和人外周血NK细胞活性均有抑制作用。100μg/ml培养液的DF_1和DF_2对PHA诱导的淋转抑制率分别为41.7%和68.2%;对NK细胞活性的抑制率分别为30.4%和81.5%。在PHA诱导的淋转中同时加入DF_1和DF_2,抑制率为80.7%。比较不同浓度的DF_1,DF_2对PHA诱导的淋转抑制作用结果表明:100μg/ml具有相对高的抑制活性。不同浓度的DF_2对NK活性抑制作用也表现出类似结果。
This article describes the whole process of human seminal plasma immunosuppressive factor isolation and immunoassay. Two immunosuppressive factors, DF_1 and DF_2, were isolated from human seminal plasma by low temperature and high speed centrifugation, Sephadex G-100 gel filtration and DEAE ion exchange chromatography. DF_2 reached polyacrylamide gel electrophoresis pure, was a single protein band, DF_1 showed two protein bands. Both of them inhibit PHA-induced human peripheral blood lymphocyte transformation and human peripheral blood NK cell activity. The inhibitory rates of DF_1 and DF_2 in 100μg / ml culture medium on PHA-induced lymph node metastasis were 41.7% and 68.2%, respectively. The inhibitory rates on NK cell activity were 30.4% and 81.5%, respectively. DF_1 and DF_2 were added simultaneously in PHA-induced lymph node metastasis. The inhibition rate was 80.7%. Comparing different concentrations of DF_1, DF_2 induced PHA-induced lymph node metastasis results show that: 100μg / ml has a relatively high inhibitory activity. Different concentrations of DF_2 inhibit NK activity also showed similar results.