IL1 RECEPTOR ANTAGONIST--INTRACELLULAR VARIANTS

IL1 受体拮抗剂——细胞内变异体

• 批准号: 2683284
• 负责人: WILLIAM P AREND
• 金额: $ 22.86万
• 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/2683284
• 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.

本研究的总体目标是进一步