Pathogenesis of Fever in Man

项目摘要/摘要 白介素38(IL-38)是白介素1家族中的一员，但直到最近才被发现 20年前。它是一种被忽视的细胞因子，因为没有发现IL-38如何发挥作用的受体。 然而，在2012年，我们报道了重组人IL-38与IL-36受体(现在的IL-1R6)和 抑制IL-17和IL-22的产生。在那项研究中，我们提出IL-38作为一种受体 IL-1R6拮抗剂。但IL-38的剂量效应不是受体拮抗剂，而是受体拮抗剂。 起到了抑制细胞活动的作用。新的数据表明，IL-38需要IL-1R6，一种在体内的孤儿受体 IL-1家族，抑制IL-17。以前被称为IL-1受体相关蛋白Like-1，IL-1R9将是 研究其在重组IL-38抑制天然免疫中的可能作用。使用CRISPR/CAS 方法，我们已经产生了一个IL-38缺乏的小鼠群体。这些老鼠被用来确定 人类炎症性疾病小鼠模型对内源性IL-38的需求。在这些模型中 如果IL-38缺陷小鼠的疾病严重程度恶化，我们将使用重组IL-38治疗小鼠 抑制先天炎症。为了充分了解IL-38在先天免疫中的作用，我们 产生了一个缺乏IL-1R9的小鼠克隆。我们将研究IL-1R9对血管内皮细胞功能的要求 重组IL-38在使用IL-38治疗可显著降低疾病的小鼠模型中的作用 严肃性。这一应用的一个独特方面是IL-1R9位于X染色体上，这是在 细胞因子生物学。因为IL-1R9在X染色体上，我们可以解决如何抑制先天 与女性相比，男性的免疫力受到影响。大多数自身免疫性疾病有70% 女性的偏好和每一种自身免疫性疾病都有炎症性贡献，我们有 为比较纯合子IL-1R9缺陷男性和纯合子IL-1R9女性而设计的研究。 在这些研究中，我们还将评估IL-38抑制NLRP3炎症体激活的作用 使用一种目前用于治疗患者的特定口服NLRP3抑制剂。除AIM 1和AIM 2研究外 关于重组IL-38对天然免疫的抑制和IL-1R9的可能作用，我们将产生和 检测IL-38-Fc融合蛋白(AIM3)。生产IL-38-Fc融合蛋白的基本原理是提供 IL-38疗法的临床前数据。在AIM 4中，我们解决了细胞释放IL-38的问题。IL-38 在健康受试者中循环，但在有心血管事件风险的受试者中水平明显较低。 然而，IL-38是一种B细胞产物，这表明对IL-38前体的处理和从 细胞不是通过传统的途径。我们将研究其他成员使用的分泌途径 IL-1家族：抑制NLRP3和抑制钙调蛋白。这些研究的总体目标是 白介素38的生物学和临床意义及其抗炎作用的研究进展 IL-38作为治疗剂。

Project Summary/Abstract Interleukin-38 (IL-38), a member of the IL-1 family, has not been studied until recently despite its discovery 20 years ago. It is a neglected cytokine because no receptor was identified for how IL-38 functioned. However, in 2012, we reported that recombinant human IL-38 bound to the IL-36 receptor (now IL-1R6) and suppressed the production of IL-17 and IL-22. In that study, we proposed that IL-38 acted as a receptor antagonist for IL-1R6. However, the dose-response of IL-38 did not behave as receptor antagonist but rather acted as an inhibitor of cell activities. New data suggests that IL-38 requires IL-1R6, an orphan receptor in the IL-1 Family, to suppress IL-17. Formerly termed IL-1 Receptor Associated Protein Like-1, IL-1R9 will be studied for its putative role in the suppression of innate immunity by recombinant IL-38. Using CRISPR/Cas methods, we have generated a colony of mice that are deficient in IL-38. These mice are used to determine a requirement for endogenous IL-38 in mouse models of human inflammatory diseases. In those models where disease severity worsens in IL-38 deficient mice, we will use recombinant IL-38 to treat mice for suppression of innate inflammation. In order to fully understand the role of IL-38 in innate immunity, we generated a mouse colony deficient in IL-1R9. We will study the requirement of IL-1R9 for the function of recombinant IL-38 in those mouse models where treatment with IL-38 has significantly reduced disease severity. A unique aspect of this application is that IL-1R9 in on the X-chromosome, an unusual finding in cytokine biology. Because IL-1R9 is on the X-chromosome, we can address how suppression of innate immunity is affected in males compared to females. With most autoimmune diseases having a 70% predilection for females and with each autoimmune disease there is an inflammatory contribution, we have designed studies for comparisons of homozygous IL-1R9 deficient males to homozygous IL-1R9 females. In these studies, we will also evaluate the role of IL-38 to inhibit the activation of the NLRP3 inflammasome using a specific oral NLRP3 inhibitor presently used to treat patients. In addition to AIM 1 and AIM 2 studies on recombinant IL-38 suppression of innate immunity and the putative role of IL-1R9, we will produce and test an IL-38-Fc fusion protein (AIM 3). The rationale for producing an IL-38-Fc fusion protein is to provide pre-clinical data for an IL-38 therapeutic. In AIM 4 we address the issue of IL-38 release from the cell. IL-38 circulates in healthy subjects but levels are significantly low in subjects at risk for a cardiovascular events. However, IL-38 being a B-cell product suggests that processing of the IL-38 precursor and release from the cell is not via traditional pathways. We will examine pathways for secretion that are used by other members of the IL-1 Family : inhibition of NLRP3 and inhibition of calpains. The overall goal of these studies is to advance the biology and clinical significance of IL-38 as well as to exploit the anti-inflammatory properties of IL-38 as a therapeutic.