IL-1 signaling in defense against Aspergillus fumigatus infection

项目总结/摘要 侵袭性曲霉病（IA）是一种危及免疫功能低下患者生命的肺部感染， 烟曲霉虽然以前的工作已经确定了招募的吞噬细胞作为关键的细胞球员， 控制A.在烟曲霉感染中，关于肺结构细胞在肺结核中的作用存在一个主要的知识缺口。 建立肺部的抗真菌环境。我的初步数据表明，IL-1受体（IL-1 R1） 信号传导促进嗜中性粒细胞和单核细胞杀伤A.烟曲霉孢子（称为分生孢子）和IL-1 R1 在肺结构细胞上的表达是产生关键的抗真菌细胞因子如GM-CSF所必需的。这些 数据有力地提示了结构细胞在A.感染，虽然 IL-1 β/β-产生细胞和IL-1 R1表达细胞的同一性尚不清楚。我假设白细胞介素的产生- 分别由募集的和肺驻留的吞噬细胞产生的1 μ M和1 μ M，是最佳的免疫细胞杀伤所必需的。 Af分生孢子和小鼠存活，并且结构细胞的IL-1 R1表达是广泛产生 抗真菌细胞因子通过不同的结构细胞亚群。我将通过以下具体目标来检验这一假设： 目的1：探讨IL-1 β/β的细胞来源及其对预防死亡和预后的作用。 A.烟熏感染的肺我将在受感染的细胞中鉴定IL-1 β/β产生细胞， 使用ELISA、流式细胞术和免疫荧光显微镜，在野生型小鼠和 耗尽候选生产细胞类型的小鼠。我将用荧光A感染缺乏IL-1 β或IL-1 β的小鼠。 烟曲霉报告子来测试每种细胞因子对真菌杀伤和鼠存活的贡献。 目的2：定义产生调节质量的关键细胞因子的IL-1响应性结构细胞类型的图谱 和抗A抗体的量。烟曲霉免疫反应我将鉴定表达IL-1 R1的结构细胞亚群 用流式细胞术和显微镜观察肺内的细胞。我将测试特定细胞亚群对IL-1 R1的需求 通过使用在三种不同结构细胞上具有条件性Il 1 r1缺失的小鼠， 子集我将测试结构细胞上的IL-1 R1是否是抗真菌细胞因子产生所必需的，并确定 哪些结构细胞可以产生这些细胞因子。 在进行这些目标，我将确定结构和免疫细胞参与IL-1 R1依赖性 A期间的串扰。烟曲霉感染这些结果将促进我们对免疫的理解，以医学- 重要真菌病原体。我设计了这个培训计划，用于真菌发病机理的广泛培训， 细胞免疫学申办者的实验室和主办机构是开展 这种培训有助于在研究和培训的其他方面取得成功的可能性很高。

PROJECT SUMMARY/ABSTRACT Invasive aspergillosis (IA) is a life-threatening pulmonary infection of immunocompromised patients caused by Aspergillus fumigatus. While previous work has identified recruited phagocytes as critical cellular players in controlling A. fumigatus infection, a major knowledge gap exists regarding the role of lung structural cells in establishing the anti-fungal environment of the lung. My preliminary data indicates that IL-1 receptor (IL-1R1) signaling promotes neutrophil and monocyte killing of A. fumigatus spores (called conidia) and that IL-1R1 expression on lung structural cells is required for production of key anti-fungal cytokines such as GM-CSF. These data strongly suggest an accessory immune role for structural cells during A. fumigatus infection, although the identity of IL-1/-producing cells and IL-1R1-expressing cells is unknown. I hypothesize that production of IL- 1 and  by recruited and lung-resident phagocytes, respectively, is required for optimal immune cell killing of Af conidia and murine survival and that IL-1R1 expression by structural cells is required for broad production of anti-fungal cytokines by diverse structural cell subsets. I will test this hypothesis with the following specific aims: Aim 1: To investigate the cellular sources of IL-1/ and their contributions to prevention of mortality and efficiency of conidial killing in the A. fumigatus-infected lung. I will identify IL-1/ producing cells in infected mouse lungs using ELISAs, flow cytometry, and immunofluorescence microscopy, in both wild-type mice and mice depleted of candidate producer cell types. I will infect mice deficient in IL-1 or  with a fluorescent A. fumigatus reporter to test the contributions of each cytokine to fungal killing and murine survival. Aim 2: To define a map of IL-1 responsive structural cell types that produce key cytokines modulating the quality and quantity of the anti-A. fumigatus immune response. I will identify IL-1R1-expressing structural cell subsets in the lung using flow cytometry and microscopy. I will test the requirement for IL-1R1 on specific cellular subsets for murine survival and fungal killing by using mice with conditional Il1r1 deletion on three different structural cell subsets. I will test whether IL-1R1 on structural cells is required for anti-fungal cytokine production and determine which structural cells may produce these cytokines. Upon conducting these aims, I will determine the structural and immune cells involved in IL-1R1-dependent crosstalk during A. fumigatus infection. These results will advance our understanding of immunity to a medically- important fungal pathogen. I have designed this training plan for broad training in fungal pathogenesis and cellular immunology. The sponsor's laboratory and the host institution are the ideal environment for conducting this training and contribute to a high likelihood of success in both research and other aspects of training.