Innate immunity to adenovirus

产品说明：腺病毒载体（Ad）是美国临床试验中第二常用的载体，用于治疗许多先天性和获得性人类疾病。尽管野生型Ad的自然感染在人类中基本上是无害的，但用于基因递送目的的静脉内Ad施用，特别是在高剂量下，刺激强烈的先天免疫和炎症宿主应答。使用完全体内方法和大量基因缺陷小鼠模型，PI的实验室发现血液中的Ad载体结合凝血因子，促进先天免疫受体识别病毒并激活IL-1家族细胞因子，进而触发先天免疫和全身炎症的细胞和分子效应子级联反应。最近，我们发现，广告管理触发一种新的坏死样类型的细胞死亡的肝脏住宅巨噬细胞，枯否细胞，这是机械控制的干扰素调节因子3，IRF 3。重要的是，除了Ad载体之外，在将细菌病原体单核细胞增生李斯特菌和完全合成的纳米颗粒血管内递送到小鼠中之后也观察到这种快速坏死的枯否细胞死亡。我们在播散性感染动物模型中的初步研究表明，IRF 3依赖性巨噬细胞坏死是高度促炎性的，并导致白细胞减少症，这是全身炎症的临床体征。该资助提案的主要目标是i）定义激活IRF 3以在体内执行坏死巨噬细胞死亡的分子机制和信号伴侣，以及ii）定义在新型人源化小鼠模型中对基因转移载体的先天免疫和炎症反应。这些目标将通过以下具体目标实现：1。鉴定IRF 3修饰的分子特征，其与其作为体内应答Ad的坏死库普弗细胞死亡的触发因子的功能相关。2.鉴定靶向IRF 3的信号通路的组分，以在体内执行坏死性库普弗细胞死亡。3.在新型人源化小鼠模型中确定先天免疫激活和对基因转移载体的全身先天免疫和炎症应答的特征。从拟议的研究中获得的概念和机制的进步有可能大大提高我们的理解，广告与宿主细胞在体内的相互作用，提供了新的见解基本的细胞和分子生物学过程相关的促炎抗病原体反应，并将允许优化基因转移载体的疫苗接种和基因转移应用。

DESCRIPTION: Adenovirus vectors (Ad) are the second most frequently used vectors in clinical trials in the US to treat numerous inborn and acquired human diseases. Even though natural infections with wild-type Ad are largely harmless in humans, an intravenous Ad administration for gene delivery purposes, especially at high doses, stimulates strong innate immune and inflammatory host responses. Using an entirely in vivo approach and a large collection of gene-deficient mouse models, the PI's laboratory has discovered that Ad vectors in the blood bind coagulation factors that facilitate virus recognition by innate immune receptors and activate IL-1 family cytokines, which, in turn, trigger a cascade of cellular and molecular effectors of innate immunity and systemic inflammation. Most recently, we found that Ad administration triggers a novel necrosis-like type of cell death of liver residential macrophages, Kupffer cells, that is mechanistically controlled by interferon regulatory factor 3, IRF3. Importantly, in addition to Ad vectors, this rapid necrotic Kupffer cell death was also observed after intravascular delivery into mice of both the bacterial pathogen L.monocytogenes and fully synthetic nanoparticles. Our preliminary studies in animal models of disseminated infections revealed that IRF3- dependent macrophage necrosis is highly pro-inflammatory and contributes to leukocytopenia, a clinical sign of systemic inflammation. The major goals of this grant proposal are i) to define the molecular mechanism and signaling partners that activate IRF3 to execute necrotic macrophage cell death in vivo and ii) to define the innate immune and inflammatory responses to gene transfer vectors in a novel humanized mouse model. These goals will be achieved via the following Specific Aims: 1. To identify the molecular signature of IRF3 modification(s) associated with its function as a trigger of necrotic Kupffer cell death in response to Ad in vivo. 2. To identify components of the signaling pathway that target IRF3 for execution of necrotic Kupffer cell death in vivo. 3. To define the signature of innate immune activation and systemic innate immune and inflammatory responses to gene transfer vectors in a novel humanized mouse model. The conceptual and mechanistic advances obtained from the proposed studies have the potential to greatly improve our understanding of Ad interactions with host cells in vivo, provide new insights into fundamental cell and molecular biological processes related to pro-inflammatory anti-pathogen responses, and will allow for optimization of gene transfer vectors for vaccination and gene transfer applications.