IL1 RECEPTOR ANTAGONIST--INTRACELLULAR VARIANTS

这项拟议研究的总体目标是进一步 描述生产机制以及可能产生的影响 特异性IL-1的两种结构变异体的细胞功能 在细胞内发现受体拮抗剂(IL-1ra)。它的第一种形式 分子，称为分泌物或sIL-1ra，是一种主要的胞外产物 单核细胞、巨噬细胞和中性粒细胞。第二种形式是iclL-1Ra， 角质形成细胞和其他上皮细胞中发现的一种细胞内分子 细胞。我们现在已经确定，iclL-1Ra也是 滑膜成纤维细胞，由单核细胞延迟合成 和巨噬细胞。在最近的研究中，我们观察到16kD的低 IL-1ra以相对分子质量形式存在于 单核细胞、巨噬细胞系和中性粒细胞。的术语 这一独特的分子家族已经扩展到：SLL-1Ra，iclL- 1Ra(1)(原始胞质形式)和iclL-1Ra(2)(新低 分子量胞质形式)。 需要检验的假设是两种细胞内物质的产生 IL-1ra在人单核细胞、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国 单核巨噬细胞产生sIL-1ra的机制比较 或者成纤维细胞。细胞内iclL-1Ra(1)或iclL-1Ra(2)的存在 可能影响IL-1受体表达的调节或特异性 IL-1诱导的生物学反应。 这些研究将涉及三个具体目标：1)确定 IclL-1Ra产生的调节机制(1)。2)至 确定iclL-1Ra(2)的生产调节机制。3)至 确定体内发现的LL-1Ra结构变体的影响 细胞对IL-1受体表达的调控或对特异性IL-1的调控 诱导的生物反应。 这些研究将利用聚合酶链式反应技术 和用于IL-1ra的引物，特异性ELISA，转染，形态 方法和均衡约束。这些研究都与人类有关 IL-1炎症作用重要的关节和皮肤病 在病理生理学方面。L-1ra不仅能抑制IL-1与细胞因子的结合 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的效应 信号的表达或启动。拟议的研究将进一步 确定细胞特定功能的产生机制。 对LL-1ra的生理和作用有更深入的了解 带来了独特和新颖的方法来治疗患者 患有急慢性关节炎或皮肤病。

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.