Neifeld J P, Lippman M E, Tormey D C
J Biol Chem. 1977 May 10;252(9):2972-7.
The presence of specific steroid hormone receptors in human lymphocytes was investigated in unstimulated and phytohemagglutinin-stimulated glass wool column-purified peripheral blood lymphocytes. Specific steroid binding in intact cells was determined by a whole cell competitive binding assay. Non-phytohemagglutinin-stimulated lymphocytes had about 2700 specific glucocorticoid binding sites per cell; phytohemagglutinin stimulation induced a 2 to 3-fold increase in glucocorticoid receptor activity within 16 h of culture. No estrogen, androgen, or progestin binding sites were detected in either unstimulated or phytohemagglutinin-stimulated peripheral blood lymphocytes. Scatchard analysis of glucocorticoid binding was consistent with a single class of receptor sites with a dissociation constant (Kd) of about 5.5 x 10(-9) M (correlation coefficient r = -0.96). Significant competition for radiolabeled dexamethasone binding was not observed with steroids lacking glucocorticoid activity. There was good agreement between relative binding affinities of various steroids to glucocorticoid receptor in lymphocytes and ability of these steroids to inhibit phytohemagglutinin-stimulated thymidine incorporation.
在未受刺激和经植物血凝素刺激的玻璃棉柱纯化外周血淋巴细胞中,研究了人淋巴细胞中特定甾体激素受体的存在情况。通过全细胞竞争结合试验测定完整细胞中的特异性甾体结合。未受植物血凝素刺激的淋巴细胞每个细胞约有2700个特异性糖皮质激素结合位点;植物血凝素刺激在培养16小时内使糖皮质激素受体活性增加2至3倍。在未受刺激或经植物血凝素刺激的外周血淋巴细胞中均未检测到雌激素、雄激素或孕激素结合位点。糖皮质激素结合的Scatchard分析与一类解离常数(Kd)约为5.5×10(-9)M的受体位点一致(相关系数r = -0.96)。缺乏糖皮质激素活性的甾体未观察到对放射性标记地塞米松结合的显著竞争。淋巴细胞中各种甾体与糖皮质激素受体的相对结合亲和力与这些甾体抑制植物血凝素刺激的胸腺嘧啶核苷掺入的能力之间存在良好的一致性。