人类精浆免疫抑制因子的研究——Ⅰ.分离和免疫活性测定

STUDY OF IMMUNOSUPPRESSIVE FACTORS IN HUMAN SEMINAL PLASMA——Ⅰ. ISOLATION AND DETERMINATION OF IMMUNE ACTIVITIES

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。

The isolation procedures of immunosuppressive factors (IFs) in human seminal plas-ma (SP), as well as the assays for their immunosuppressive effects, were reported. Twotypes of IFs-DF_1, DF_2 were obtained through high speed centrifugation at low temperature,Sephadex G-100 gel filtration, and DEAE-cellulose ion-exchange chromatography. The DF_2showed a single band in polyacrylamide gel electrophoresis, while the DF_1 appearcd as twobands. Both DF_2 and DF_1 had inhibitory effect in two assay systems of cell-mediated im-munity. Their inhibitory rates for PHA—induced lymphocyte blastogenesis when a concen-tration of 100μg/ml exposed were 68.2%, 41.7%, respectively. The inhibitory ratesfor NK activity were 81.5%, 30.4%, respectively with same concentration. In caseboth DF_1 and DF_2 were added simultaneously to the PHA-induced lymphocyte system, theinhibitory rate was 30.7%. The comparison of inhibitory rates of different concentrationsof DF_1, DF_2 in PHA-induced lymphocyte blastogenesis showed that 100μg/ml would givethe maximal inhibitory effect. The different concentration of DF_2 showed a comparable re-sult in the NK activity assay.