Merz W E, Schmidt W, Lenhard V
Hoppe Seylers Z Physiol Chem. 1979 Oct;360(10):1433-44. doi: 10.1515/bchm2.1979.360.2.1433.
The effect of choriogonadotropin of different purities on the transformation of peripheral human lymphocytes was studied. Various crude hormone batches inhibited lymphocyte transformation in a dose-dependent manner, both in the mixed lymphocyte reaction and in the phytohemagglutinin-induced stimulation. The inhibitory activity, however, was found not to be correlated with the gonadotropic activity of the crude hormone batches (2660-4300 IU/mg). Choriogonadotropin (13 000 IU/mg), which was purified in 3 steps, showed no inhibitory effect except at high doses (greater than 5000 IU/ml final dilution). More detailed investigations provided evidence that in the first step of the choriogonadotropin purification procedure (batch adsorption of crude choriogonadotropin on SP-Sephadex C-50), the inhibitory activity can be enriched in a fraction (Fract. I) which displays a very low gonadotropic activity (less than 500 IU/mg). A further separation of Fract. I was achieved by isoelectric focusing as well as by chromatography on DEAE-Sephadex A-25. By these means, the inhibitory potency could be enriched more than 100-fold. The substances which display inhibition of DNA synthesis in lymphocytes were proven to act in a nontoxic way. A preliminary characterization of the strongly inhibiting substances which show a dose-dependent suppression of lymphocyte transformation by about 99%, showed that this effect is probably exerted via non-dialysable sialoglycoproteins. By a fourth purification step entailing a chromatography of purified choriogonadotropin (13 000 IU/mg) on SP-Sephadex C-50, a highly purified choriogonadotropin (14000 IU/mg) could be obtained which showed no inhibitory effect on lymphocyte transformation (in both mixed lymphocyte reaction and in phytohemagglutinin-induced stimulation) up to a dose of 43 000 IU/ml. The components which were removed from choriogonadotropin in this step seem to be immunologically identical with the strongly inhibiting substances isolated by isoelectric focusing. These investigations demonstrate that biologically active, highly purified choriogonadotropin is unable to inhibit lymphocyte transformation. The inhibitory activity of crude hormone can be enriched in choriogonadotropin-free fractions. Therefore, it is concluded that the inhibitory activity of crude hormone is not a property of choriogonadotropin itself.
研究了不同纯度的绒毛膜促性腺激素对人外周血淋巴细胞转化的影响。在混合淋巴细胞反应和植物血凝素诱导的刺激中，各种粗制激素批次均以剂量依赖性方式抑制淋巴细胞转化。然而，发现抑制活性与粗制激素批次的促性腺活性（2660 - 4300 IU/mg）无关。经过三步纯化的绒毛膜促性腺激素（13000 IU/mg），除高剂量（最终稀释度大于5000 IU/ml）外无抑制作用。更详细的研究表明，在绒毛膜促性腺激素纯化过程的第一步（将粗制绒毛膜促性腺激素分批吸附到SP - Sephadex C - 50上），抑制活性可富集在一个促性腺活性非常低（小于500 IU/mg）的组分（组分I）中。通过等电聚焦以及在DEAE - Sephadex A - 25上进行色谱分离进一步分离了组分I。通过这些方法，抑制效力可富集100倍以上。已证明在淋巴细胞中抑制DNA合成的物质以无毒方式起作用。对强烈抑制物质的初步表征表明，其以剂量依赖性方式使淋巴细胞转化抑制约99%，这种作用可能是通过不可透析的唾液糖蛋白发挥的。通过第四步纯化步骤，即将纯化的绒毛膜促性腺激素（13000 IU/mg）在SP - Sephadex C - 50上进行色谱分离，可获得高度纯化的绒毛膜促性腺激素（14000 IU/mg），其在剂量高达43000 IU/ml时对淋巴细胞转化（在混合淋巴细胞反应和植物血凝素诱导的刺激中）均无抑制作用。在这一步中从绒毛膜促性腺激素中去除的组分在免疫学上似乎与通过等电聚焦分离的强烈抑制物质相同。这些研究表明，具有生物活性的高度纯化的绒毛膜促性腺激素不能抑制淋巴细胞转化。粗制激素的抑制活性可在不含绒毛膜促性腺激素的组分中富集。因此，得出结论，粗制激素的抑制活性不是绒毛膜促性腺激素本身的特性。
