MODE OF ACTION OF STEROID HORMONES

这项研究的长期目标是了解 糖皮质激素在分子、细胞和免疫系统中的作用 生理水平。免疫系统的细胞、组织和功能 是糖皮质激素治疗作用的主要靶点，如 免疫抑制和抗炎药物，以及在治疗 淋巴细胞性白血病和淋巴瘤。 具体目标针对三个基本问题：(一) 伴随“激活”的糖皮质激素受体的结构变化 (类固醇-受体复合体获得亲和力的转化 对于细胞核和DNA)；(Ii)糖皮质激素诱导的mRNAs和 大鼠胸腺细胞中的蛋白质与葡萄糖的快速作用 转运；(Iii)糖皮质激素和干扰素的相互作用 完整小鼠的免疫过程。 (I)假设细胞中的糖皮质激素受体是周期性的 磷酸化和去磷酸化，这是从早期的工作中出现的 这个项目将通过测量受体的32P标记来进行测试 用多种方法处理WEHI-7小鼠胸腺瘤细胞。纯化的受体 将通过以下方式分析32P的含量和位置 多肽图谱，这也将被单独应用于识别多肽 DNA结合域的。抗体将被用来识别 在缺乏能量的细胞中，受体的非结合形式。一种动力学模型 受体循环将用糖皮质激素拮抗剂进行测试。 (Ii)糖皮质激素抑制孵育大鼠胸腺的葡萄糖转运 细胞数在20分钟内，1小时后起其他作用。有证据表明 这些效应是由诱导的编码蛋白质的mRNAs介导的 抑制葡萄糖转运等。消减杂交将用于 罕见的糖皮质激素诱导的胸腺信使核糖核酸的鉴定及其表达 各自的蛋白质。诱导蛋白将分别由其各自的 推测具有抑制葡萄糖转运蛋白的能力。 (3)糖皮质激素对伽马的许多潜在重要影响 干扰素的产生和作用已在早期的工作中得到证实 这个项目与文化系统有关。很少有在动物身上得到证实的。 为了弥合培养系统和整个动物之间的差距，小鼠将被 用于研究肾上腺切除术、外源性干扰素和 糖皮质激素对Ig G-Fc受体表达、B细胞(抗体)反应、 和干扰素基因水平。

The long-term objective of this research is an understanding of actions of glucocorticoids on the immune system at molecular, cellular and physiological levels. Cells, tissues and functions of the immune system are major targets of glucocorticoids in their therapeutic functions as immunosuppressive and anti-inflammatory agents, and in the treatment of lymphocytic leukemias and lymphomas. Specific aims are directed towards three fundamental problems: (i) structural changes in glucocorticoid receptors that accompany "activation" (the transformation by which steroid-receptor complexes acquire affinity for nuclei and DNA); (ii) induction by glucocorticoids of mRNAs and proteins in rat thymus cells, in relation to rapid effects on glucose transport; (iii) interactions of glucocorticoids and gamma interferon on immune processes in intact mice. (i) The hypothesis that glucocorticoid receptors in the cell are cyclically phosphorylated and dephosphorylated, which emerged from earlier work on this project, will be tested by measuring 32P labeling of receptors in WEHI-7 mouse thymoma cells treated in various ways. Receptors purified with a monoclonal antibody will be analyzed for 32P content and location by peptide mapping, which will also be applied separately to identify peptides of the DNA-binding domain. The antibody will be used to identify nonbinding forms of receptors in energy-deprived cells. A kinetic model of receptor cycling will be tested with glucocorticoid antagonists. (ii) Glucocorticoids inhibit glucose transport in incubated rat thymus cells within 20 min, and exert other effects by 1 hour. Evidence suggests these effects are mediated by induced mRNAs coding for proteins that inhibit glucose transport, etc. Subtractive hybridization will be used to identify rare glucocorticoid-induced species of thymus mRNA and their respective proteins. Induced proteins will separately be sought by their putative ability to inhibit glucose transporters. (iii) Many potentially important effects of glycocorticoids on gamma interferon production and actions have been demonstrated in earlier work on this project with culture systems. Few have been demonstrated in animals. To bridge the gap between culture systems and whole animals, mice will be used to study the effects of adrenalectomy, exogenous gamma interferon and glycocorticoids on IgG-Fc receptor expression, B-cell (antibody) responses, and gamma interferon mRNA levels.