ROLE OF ENDOGENOUS IL1 RECEPTOR ANTAGONIST

白细胞介素-1（IL-1）与脑血管病的发病机制有关， 感染和其他炎性疾病。 IL-1受体拮抗剂 （IL-1 Ra）是天然存在的IL-1活性抑制剂， 与IL-1竞争细胞表面受体的占有， 无激动剂活性。 IL-1 Ra可以作为分泌蛋白（sIL-2 R）被发现。 1 Ra），其主要由单核细胞产生，而角质形成细胞 产生细胞内形式的IL-1 Ra（icIL-1 Ra）。 IL-1和IL-10两者均 1 Ra的产生在炎症过程中增加。 人们日益 有证据表明外源性给予IL-1 Ra可降低 疾病的各种模型，但有一个缺乏研究 检查内源性IL-1 Ra在炎症中的作用。 的 这项拨款中提出的实验旨在研究 如果内源性IL-1 Ra的合成被反义或其 作用被抗IL-1 Ra抗体中和。 初步研究 表明抗人IL-1 Ra抗体阻止重组IL-1 Ra 抑制IL-1诱导的人血细胞因子合成 单核细胞 研究将检查中和的效果 内源性sIL-1 Ra对人细胞因子级联反应的影响 用试剂如内毒素刺激的单核细胞。 类似的实验 将使用互补于 人sIL-1 Ra。 在角质形成细胞中，对icIL-1 Ra特异性的反义将 用于确定icIL-1 Ra在IL-1诱导的增殖中的作用 和刺激诱导的细胞因子合成。 因为研究表明IL- 1可以通过直接与DNA结合来影响某些基因的转录， 将确定icIL-1 Ra水平的降低是否与 反义icIL-1 Ra增加IL-1从细胞表面的移位 到细胞核。 IL-1 Ra在体内的作用也将通过 检测反义icIL-1 Ra对注射小鼠存活率的影响 内毒素。 为了识别触发icIL-1 Ra的刺激， 启动子，上皮细胞将用icIL-1 Ra转染 与氯霉素乙酰转移酶基因连接的启动子。 这些 研究将icIL-1 Ra的产生调节与细胞事件联系起来 由炎症刺激引起的。

Interleukin-1 (IL-1) has been implicated in the pathogenesis of infection and other inflammatory diseases. IL-1 receptor antagonist (IL-1Ra) is a naturally occurring inhibitor of IL-1 activity that competes with IL-1 for occupancy of cell-surface receptors but possesses no agonist activity. IL-1Ra can be found as a secreted protein (sIL- 1Ra) which is primarily produced by monocytes, whereas keratinocytes produce an intracellular form of IL-1Ra (icIL-1Ra). Both IL-1 and IL- 1Ra production are increased during inflammation. There is increasing evidence that exogenously administered IL-1Ra reduces the severity of disease in a variety of models, but there is a paucity of studies examining the role of endogenous IL-1Ra in inflammation. The experiments proposed in this grant are designed to examine what happens if the synthesis of endogenous IL-1Ra is inhibited by antisense or its actions are neutralized by anti-IL-1Ra antibodies. Preliminary studies indicate that anti-human IL-1Ra antibody prevents recombinant IL-1Ra from inhibiting IL-1-induced cytokine synthesis in human blood mononuclear cells. Studies will examine the effect of neutralizing endogenous sIL-1Ra on the cascade of cytokines synthesized in human monocytes stimulated with agents such as endotoxin. Similar experiments will be performed using antisense oligodeoxynucleotides complementary to human sIL-1Ra. In keratinocytes, antisense specific for icIL-1Ra will be used to determine the role of icIL-1Ra in IL-1-induced proliferation and stimulus-induced cytokine synthesis. Since studies suggest that IL- 1 can affect the transcription of some genes by binding directly to DNA, it will be ascertained whether a reduction of icIL-1Ra levels with antisense icIL-1Ra increases translocation of IL-1 from the cell-surface to the nucleus. The role of IL-1Ra in vivo will also be assessed by examining the effect of antisense icIL-1Ra on survival in mice injected with endotoxin. To identify stimuli which trigger the icIL-1Ra promoter, epithelial cells will be transfected with the icIL-1Ra promoter linked to the chloramphenicol acetyltransferase gene. These studies will link regulation of icIL-1Ra production to cellular events induced by inflammatory stimuli.