ROLE OF INTERLEUKIN-1 INHIBITOR--INFLAMMATORY SYNOVITIS

我的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和炎症性疾病中的作用。一种特定的 在过去的几年里，人们对白细胞介素1的调节一直很感兴趣 (IL-1)人单核细胞产生。在这些研究的过程中 观察到单核细胞培养在贴壁的免疫球蛋白底物上。 释放IL-1抑制活性。这种IL-1抑制剂(IL-1-3inh)是 =25kD的大小，并特异性地阻断IL-1对两种免疫系统的刺激 和炎性细胞。揭示了该IL-1-inh的作用机制。 抑制IL-1α和β受体结合。 在这些拟议的实验中要检验的假设是，IL 人单核细胞分泌α/β和IL-1-inh受到调控 分开的。这一假设将在五个具体目标下进行检验：1) 以确定IL-1和IL-1-inh是否由 2)检测IL-1-inh的产生及IL-1-inh的mRNA水平。 来自正常供者或关节炎患者的体外来源的巨噬细胞 从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底物对调控的影响 IL-1-inh的产生；4)确定IL的调节机制 人单核细胞和巨噬细胞产生1-异烟肼；以及5)检测 重组人IL-1-inh与多种人体细胞的结合特性 拟在这些研究中使用的技术包括 细胞内IL-1α、IL-1β和IL-1-inh蛋白水平的测定 用特定的酶联免疫吸附试验测定裂解物和上清液 体外杂交法测定相对稳态mRNA水平。其他内容 技术将包括免疫细胞化学定位，原位 杂交、核延伸、信使核糖核酸稳定性和体外翻译。 抗IL-1-inh抗体、cDNA探针和重组IL-1-inh 合作者已经向我提供了分子，用于这些 实验。 IL-1被认为是一种重要的组织破坏介质。 人类的各种疾病。一种专门阻断受体的分子 IL-1的结合可能在这些疾病中具有治疗潜力。这些 拟议的研究应提供有关以下机制的重要信息 人单核细胞产生IL-1抑制物的调节 滑膜巨噬细胞。

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.