IL1 RECEPTOR ANTAGONIST--INTRACELLULAR VARIANTS

IL1 受体拮抗剂——细胞内变异体

• 批准号: 2079858
• 负责人: WILLIAM P AREND
• 金额: $ 21.09万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-31 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2079858
• 关键词: RNA splicing; cell type; cytokine receptors; enzyme linked immunosorbent assay; genetic promoter element; genetic translation; glycoprotein biosynthesis; glycoprotein structure; human tissue; immunocytochemistry; inflammation; inhibitor /antagonist; interleukin 1; intracellular transport; macrophage; monocyte; protein isoforms; protein structure function; receptor binding; receptor expression; tissue /cell culture

The overall objective of this proposed research is to further characterize the mechanisms of production, and the possible influences on cell function, of two structural variants of the specific IL-1 receptor antagonist (IL-1ra)found inside cells. The first form of this molecule, termed secretory or slL-1ra, is a major extracellular product of monocytes, macrophages and neutrophils. The second form is iclL-1ra, an intracellular molecule found in keratinocytes and other epithelial cells. We have now determined that iclL-1ra is also a major product of synovial fibroblasts and is synthesized in a delayed fashion by monocytes and macrophages. In recent studies, we have observed that a 16kD low molecular weight form of IL-1ra is present in considerable amounts in monocytes, macrophage cell lines and neutrophils. The terminology for this unique family of molecules has been expanded to: slL-1ra, iclL- 1ra(1)(the original cytosolic form), and iclL-1ra(2) (the new low molecular weight cytosolic form). The hypothesis to be examined is that the production of two intracellular forms of IL-1ra by human monocytes, macrophages, fibroblasts and keratinocytes, may involve unique transcriptional or transnational mechanisms in comparison to slL-1ra production by mononuclear phagocytes or fibroblasts. The presence of iclL-1ra(1) or iclL-1ra(2) inside cells may influence the regulation of lL-1 receptor expression or particular lL-1-induced biological responses. Three specific aims will be addressed in these studies: 1) To determine the mechanisms of regulation of production of iclL-1ra(1). 2) To determine mechanisms of regulation of production of iclL-1ra(2). 3) To determine the influence of structural variants of lL-1ra found inside cells on regulation of lL-1 receptor expression or on specific lL-1 induced biological responses. These studies will utilize the techniques of polymerase chain reaction and primers for lL-1ra, specific ELISA, transfection, morphological approaches, and equilibrium binding. These studies are related to human joint and skin diseases where inflammatory effects of lL-1 are important in pathophysiology. L-1ra may not only inhibit binding of lL-1 to extracellular receptors, but the intracellular structural variants of this molecule may decrease lL-1 effects through influencing receptor expression or initiation of signals. The proposed studies will further determine the mechanisms of production to specific functions of the cell. A greater understanding of the physiology and effects of lL-1ra should result in unique and novel ways to approach the treatment of patients with acute and chronic arthritis or skin diseases.

这项研究的总体目标是进一步 描述生产机制和可能的影响 对细胞功能的影响， 受体拮抗剂（IL-1 ra）发现细胞内。 第一种形式 一种称为分泌型或sIL-1 ra的分子是一种主要的细胞外产物 单核细胞、巨噬细胞和中性粒细胞。 第二种形式是icIL-1 ra， 一种在角质形成细胞和其他上皮细胞中发现的细胞内分子 细胞 我们现在已经确定iclL-1 ra也是 滑膜成纤维细胞，并由单核细胞以延迟方式合成 和巨噬细胞。 在最近的研究中，我们观察到16 kD低 分子量形式的IL-1 ra以相当大的量存在于 单核细胞、巨噬细胞系和嗜中性粒细胞。 的术语 这一独特的分子家族已扩展到：sIL-1 ra，icIL-1， 1 ra（1）（原始胞质形式）和iclL-1 ra（2）（新的低水平 分子量胞质形式）。 要检验的假设是，两个细胞内的生产 人单核细胞、巨噬细胞、成纤维细胞和 角质形成细胞，可能涉及独特的转录或跨 与单核吞噬细胞产生sIL-1 ra相比的机制 或成纤维细胞。 细胞内存在iclL-1 ra（1）或iclL-1 ra（2） 可能影响IL-1受体表达的调节， IL-1诱导的生物反应。 这些研究将涉及三个具体目标：1）确定 icIL-1 ra产生的调控机制（1）. 2)到 确定icIL-1 ra产生的调节机制（2）。3)到 确定内部发现的IL-1 ra的结构变体的影响， 细胞对IL-1受体表达的调节或对特异性IL-1 引发生物反应。 这些研究将利用聚合酶链反应技术 和用于IL-1 ra、特异性ELISA、转染、形态学 方法和平衡约束。 这些研究与人类 其中IL-1的炎症作用是重要的关节和皮肤疾病 在病理生理学上。 L-1 ra不仅可以抑制IL-1与 细胞外受体，但细胞内的结构变体 该分子可能通过影响受体而降低IL-1的作用 信号的表达或启动。 拟议的研究将进一步 确定生产机制的具体功能的细胞。 更好地理解IL-1 ra的生理学和作用， 导致独特和新颖的方法来治疗患者， 急性和慢性关节炎或皮肤病。