ROLE OF INTERLEUKIN-1 INHIBITOR--INFLAMMATORY SYNOVITIS

INTERLEUKIN-1抑制剂的作用——炎症性滑膜炎

• 批准号: 3160420
• 负责人: WILLIAM P AREND
• 金额: $ 14.65万
• 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 1994-04-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3160420
• 关键词: arthritis; cell differentiation; complementary DNA; cytokine receptors; enzyme linked immunosorbent assay; genetic manipulation; genetic translation; human tissue; immunocytochemistry; immunoglobulin G; immunoregulation; in situ hybridization; inflammation; interleukin 1; interleukin 6; macrophage; macrophage activating factor; messenger RNA; migration inhibition factor; monocyte; nucleic acid probes; receptor binding; synovitis; tissue /cell culture

The long-term objective of my laboratory is to study the role of monocytes and macrophages in human autoimmune and inflammatory diseases. A specific interest over the past few years has been the regulation of interleukin 1 (IL 1) production by human monocytes. During the course of these studies it was observed that monocytes cultured on a substrate of adherent IgG released an IL 1 inhibitory activity. This IL 1 inhibitor (IL 1-3inh) was =25 kD in size and it specifically blocked IL 1 stimulation of both immune and inflammatory cells. The mechanism of action of this IL 1-inh was shown to be inhibition of receptor binding of IL 1 alpha and beta. The hypothesis to be examined in these proposed experiments is that IL 1alpha/beta and IL 1-inh production by human monocytes are regulated separately. This hypothesis will be examined under five specific aims: 1) to determine whether IL 1 and IL 1-inh are produced simultaneously by the same cells; 2) to examine IL 1-inh production and IL 1-inh mRNA levels in vitro-derived macrophages from normal donors or arthritis patients or in synovial macrophages from arthritis patients; 3) to determine the effects of monocyte differentiating agents or of culture substrates on regulation of IL 1-inh production; 4) to determine the mechanisms of regulation of IL 1-inh production in human monocytes and macrophages; and 5) to examine the characteristics of binding of recombinant IL 1-inh to various human cells. The techniques to be utilized in these proposed studies include the measurement of IL 1alpha, IL 1beta and IL 1-inh protein levels in cell lysates and supernatants using specific ELISA's, and the determination of relative steady-state mRNA levels by in vitro hybridization. Additional techniques will include immunocytochemical localization, in situ hybridization, nuclear run-on, mRNA stability and in vitro translation. Antibodies specific to the IL 1-inh, cDNA probes and recombinant IL 1-inh molecules have been made available to me by collaborators for use in these experiments. IL 1 has been implicated as an important mediator of tissue destruction in a variety of human diseases. A molecule that specifically blocks receptor binding of IL 1 may have therapeutic potential in these diseases. These proposed studies should provide important information on the mechanisms of regulation of production of an IL 1 inhibitor by human monocytes and synovial macrophages.

我实验室的长期目标是研究单核细胞的作用 和巨噬细胞在人类自身免疫性和炎症性疾病中的作用。 特定 在过去的几年里，人们的兴趣一直是调节白细胞介素1， (IL 1）通过人单核细胞产生。 在这些研究过程中 观察到在粘附IgG的基质上培养的单核细胞 释放IL 1抑制活性。 这种IL 1抑制剂（IL 1- 3 inh） =25 kD的大小，并特异性地阻断IL-1刺激两种免疫 和炎症细胞。 这种IL 1-inh的作用机制被证明是 抑制IL 1 α和β的受体结合。 在这些拟议的实验中要检验的假设是， 1 α/β和IL 1-inh的产生受到人单核细胞的调节 分开 这一假设将在五个具体目标下进行检验：1） 为了确定IL-1和IL-1-inh是否同时产生， 2）检测IL 1-inh的产生和IL 1-inh mRNA水平， 来自正常供体或关节炎患者或 关节炎患者的滑膜巨噬细胞; 3）确定影响 单核细胞分化剂或培养基质对调节 IL-1-inh的产生; 4）确定IL-1-inh的调节机制 1-人单核细胞和巨噬细胞中的inh产生;和5）检查 重组IL 1-inh与各种人细胞的结合特性。 这些拟议研究中使用的技术包括 细胞中IL 1 α、IL 1 β和IL 1-inh蛋白水平的测量 使用特异性ELISA测定裂解物和上清液， 通过体外杂交测定相对稳态mRNA水平。 额外 技术将包括免疫细胞化学定位，原位 杂交、核运行、mRNA稳定性和体外翻译。 IL 1-inh特异性抗体、cDNA探针和重组IL 1-inh 我的合作者已经提供了一些分子， 实验 IL-1被认为是组织破坏的重要介质， 各种人类疾病。 一种能特异性阻断受体的分子 IL 1的结合可能在这些疾病中具有治疗潜力。 这些 拟议的研究应提供关于 调节人单核细胞产生IL 1抑制剂， 滑膜巨噬细胞