Impact of versican deficiency on the innate immune response to influenza virus

多功能蛋白聚糖缺陷对流感病毒先天免疫反应的影响

• 批准号: 10059171
• 负责人: William A Altemeier
• 金额: $ 61.63万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-12 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10059171
• 关键词: Address; Adhesions; Agonist; Anti-Inflammatory Agents; Bacteria; Biological; Cause of Death; Cell Culture Techniques; Cells; Chemotaxis; Chondroitin Sulfate A; Chondroitin Sulfate Proteoglycan; Cues; Data; Development; Embryonic Development; Exposure to; Extracellular Matrix; Fibroblasts; Generations; Genes; Genetically Engineered Mouse; Glycosaminoglycans; Immune response; In Vitro; Infection; Inflammatory; Innate Immune Response; Interferon Type I; Interferons; Interleukin-10; Laboratories; Leukocytes; Liquid substance; Lung; Lung Inflammation; Lung infections; Modeling; Mus; Natural Immunity; Pathway interactions; Phenotype; Poly C; Poly I-C; Population; Production; Property; Proteoglycan; Public Health; Publishing; Recovery; Research; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Source; Specificity; Stromal Cells; Structure of parenchyma of lung; TCF Transcription Factor; Testing; Therapeutic; Time; Toll-like receptors; United States; Viral; Virus; Work; beta catenin; cell type; chemokine; cytokine; design; extracellular; immunoregulation; improved; influenzavirus; macrophage; migration; mouse model; novel; response; transcription factor; type I interferon receptor; versican

PROJECT SUMMARY Our published work and preliminary data using mouse models and in vitro cell cultures show that extracellular matrix (ECM) molecules such as versican accumulate in lungs following exposure to viruses and bacteria. The creation of a specialized versican-enriched ECM coincides with invasion and retention of leukocytes in lungs during the immune response to influenza virus. We have found that two cellular sources synthesize versican in response to Toll-like receptor (TLR) agonists: macrophages and fibroblasts. Studies identify two distinct signaling pathways directing versican synthesis in these two cell types - the β-catenin/T-cell factor (TCF) pathway in stromal cells such as smooth muscle cells and fibroblasts and the TLR/Trif/Type I interferon (IFN) receptor pathway in macrophages. However, the cellular specificity of these signaling pathways is not known. Using two novel strains of genetically engineered mice in which the versican gene is deleted in a cell- and time-specific manner, our published and preliminary data shows that versican has a dramatic effect on leukocyte phenotype. When treated with polyinosinic:polycytidylic acid (poly(I:C)), mice with a global deficiency in versican have decreased recovery of leukocytes in bronchoalveolar (BAL) fluid suggesting that versican is pro-inflammatory and required for leukocyte migration into lungs. In contrast, mice lacking versican in macrophages have increased recovery of leukocytes in BAL fluid suggesting that macrophage-derived versican is anti- inflammatory and suppresses leukocyte migration into lungs. Both strains of versican-deficient mice had significantly decreased recovery of Type I IFNs and IL-10 in lung tissue or BAL fluid when compared to controls. Our in vitro studies attribute this to decreased production of these anti-inflammatory cytokines by versican-deficient macrophages. The differences observed in these novel versican-deficient mice are the basis of our central hypothesis that the formation of a versican-enriched ECM by macrophages and stromal cells provides critical contextual cues and extracellular-control of the innate immune response to viral lung infection. We propose three aims to test this hypothesis. Aim 1 defines the role of versican derived from macrophages and/or Type I IFN signaling in providing extracellular-control of the innate immune response in lungs of mice exposed to influenza virus. Aim 2 determines if stromal cells and/or β- catenin/TCF signaling promote the generation of a versican-enriched ECM that provides fine-control of the innate immune response to influenza virus. Aim 3 identifies the mechanisms and signaling pathways whereby macrophage- versus fibroblast-derived versican provide contextual extracellular-control of the innate immune response. Understanding the contextual settings in which versican provides fine-control of the innate immune response to influenza virus is critical for the design of therapeutic strategies for improving the immune response to viral lung infection.

项目概要 我们发表的工作和使用小鼠模型和体外细胞培养的初步数据表明 接触病毒后，细胞外基质 (ECM) 分子（例如多功能蛋白聚糖）会在肺部积聚 和细菌。富含多功能蛋白聚糖的专门 ECM 的产生与侵袭和保留同时发生 对流感病毒的免疫反应期间肺部白细胞的变化。我们发现两个细胞 响应 Toll 样受体 (TLR) 激动剂合成多功能聚糖的来源：巨噬细胞和 成纤维细胞。研究确定了两种不同的信号通路指导这两种细胞中多功能蛋白聚糖的合成 类型 - 平滑肌细胞等基质细胞中的 β-连环蛋白/T 细胞因子 (TCF) 通路 成纤维细胞和巨噬细胞中的 TLR/Trif/I 型干扰素 (IFN) 受体途径。然而， 这些信号传导途径的细胞特异性尚不清楚。使用两种新的基因菌株 工程小鼠中，多功能蛋白聚糖基因以细胞和时间特异性方式被删除，我们发表的 初步数据表明，Versacan 对白细胞表型具有显着影响。治疗时 使用聚肌苷：聚胞苷酸（poly(I:C)），多功能蛋白聚糖全面缺乏的小鼠的数量减少了 支气管肺泡 (BAL) 液中白细胞的恢复表明多功能蛋白聚糖具有促炎作用 白细胞迁移到肺部所需的。相比之下，巨噬细胞中缺乏多功能蛋白聚糖的小鼠 BAL 液中白细胞的恢复增加表明巨噬细胞衍生的多功能蛋白聚糖具有抗 炎症并抑制白细胞迁移到肺部。两种多功能蛋白聚糖缺陷小鼠品系均具有 与相比，肺组织或 BAL 液中 I 型 IFN 和 IL-10 的回收率显着降低 控制。我们的体外研究将此归因于这些抗炎细胞因子的产生减少 多功能蛋白聚糖缺陷型巨噬细胞。在这些新型多功能蛋白聚糖缺陷小鼠中观察到的差异是 我们的中心假设的基础是巨噬细胞和富含多功能蛋白聚糖的 ECM 的形成 基质细胞提供了关键的背景线索和先天免疫反应的细胞外控制 病毒性肺部感染。我们提出三个目标来检验这一假设。目标 1 定义了 versican 的作用 源自巨噬细胞和/或 I 型 IFN 信号传导，提供先天性细胞外控制 暴露于流感病毒的小鼠肺部的免疫反应。目标 2 确定基质细胞和/或 β- 连环蛋白/TCF 信号传导促进富含多功能蛋白聚糖的 ECM 的生成，从而提供精细控制 对流感病毒的先天免疫反应。目标 3 确定机制和信号通路 由此巨噬细胞与成纤维细胞衍生的多功能蛋白聚糖提供了对 先天免疫反应。了解 versican 提供精细控制的上下文设置 对流感病毒的先天免疫反应对于设计治疗策略至关重要 改善对病毒性肺部感染的免疫反应。