The Toponome of Virus Infected Skin

摘要 病毒性皮肤感染是人类群体中发病和死亡的重要原因，数百万人感染病毒性皮肤感染。 感染痘病毒传染性软疣病毒（MCV）的全球个人，该病毒仅在 检查由宿主免疫系统作为免疫功能低下的个人目前与衰弱的全身 感染相关的痘病毒牛痘病毒（VACV）被用来免疫数亿人 在天花根除计划期间，它仍然是最广泛使用的病毒疫苗的支柱 向量。在病毒感染过程中，靶器官的形态发生显著变化， 细胞融合在病毒生命周期的进展，以及大量的各种反应的流入， 免疫细胞感染引起的炎症反应也能刺激非经典型的分化， 与通常存在于稳态的细胞不相似的驻留或浸润细胞群。 因此，这些细胞的复杂表型需要使用大的探针组（大部分是探针）。 抗体）。然而，传统的方法（显微镜，流式细胞术，蛋白质组学或基因组学）不允许 同时分析单个细胞的蛋白质表达、蛋白质的定量和拓扑定位 细胞和蛋白质相对于彼此而不损伤和混淆组织解离，或分析 单个组织的50 - 100个参数。为了完成这项调查，我们将执行非侵入性 使用Toponome成像系统分析50多个参数，Toponome成像系统是一种成像循环显微镜， 一种自动化显微镜，允许显示大分子和细胞系统的空间分辨率 在完整的组织中有数千种潜在的相互作用。我们将分析分子和细胞的变化 在使用VACV（痘病毒感染的模型）感染小鼠皮肤后的不同点的皮肤中。 我们假设多重拓扑学分析将揭示一个独特的分子和细胞特征 痘病毒皮肤感染。在目标1中，我们将优化染色方案并检查和验证感染， 定位和表型的居民和浸润细胞群体在启动，控制和 VACV皮肤感染的消退阶段。在目的2中，我们将研究两个单独的类淀粉细胞的作用， 在建立VACV病变的形态学和浸润细胞的功能方面，我们 预计从这些研究中获得的结果将成为未来RO 1或PO 1提案的基础， 受感染的、驻留的和浸润的细胞群在病毒皮肤中相互作用的机制， 感染，有可能使用临床样本来验证我们在人类中的研究。

Abstract Viral skin infections are a significant cause of morbidity and mortality in the human populations, with millions of individuals infected worldwide with the poxvirus molluscum contagiosum virus (MCV), which is only held in check by the host immune system as immunocompromised individuals present with debilitating systemic infection. The related poxvirus Vaccinia virus (VACV) was used to immunize hundreds of millions of people during the smallpox eradication program, and remains a backbone of the most widely used viral vaccine vectors. During virus infection there is a significant change in the morphology of a target organ caused by cellular fusion during progression of the virus life cycle, as well as by the influx of a large variety of responding immune cells. The inflammation produced by infection can also stimulate the differentiation of non-classical populations of resident or infiltrating cells that do not resemble cells typically present in the steady state. Therefore, the complex phenotyping of these cells requires the use of large panels of probes (mostly antibodies). However, traditional methods (microscopy, flow cytometry, proteomics or genomics) do not allow simultaneous analysis of protein expression by individual cells, quantification and topological localization of cells and proteins relative to each other without damaging and confounding tissue dissociation, or analysis of 50-100 parameters from a single tissue. In order to accomplish this investigation we will perform non-invasive analysis of 50+ parameters using a Toponome Imaging System, an imaging cycler microscope that is a fully automated microscope that allows the spatial resolution of large molecular and cellular systems displaying many thousands of potential interactions in intact tissues. We will analyze the molecular and cellular changes in the skin at different points after infection of the skin of mice with using VACV (a model for poxvirus infection). We hypothesize that multiplexed toponomic analysis will reveal a unique molecular and cellular signature poxviral skin infection. In Aim 1 we will optimize staining protocols and examine and validate infection, localization and phenotype of resident and infiltrating cell populations during the initiation, control and resolution phases of VACV dermal infection. In Aim 2 we will examine the role of two individual meyloid cell populations in establishing both the morphology of the VACV lesion and the function of infiltrating cells. We anticipate that the results gained from these studies will form the basis of a future RO1 or PO1 proposal in which the mechanisms by which infected, resident and infiltrating cell populations interact in viral skin infections, with potential for the use of clinical samples to validate out studies in humans.