The Toponome of Virus Infected Skin

病毒感染皮肤的地形图

• 批准号: 9186754
• 负责人: Christopher C Norbury
• 金额: $ 19.34万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-10 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9186754
• 关键词: Acute; Address; Affect; Africa; Antibodies; Antiviral Agents; Area; Bone Marrow; CD8B1 gene; Cell Lineage; Cells; Clinical; Code; Complex; Data; Dendritic Cells; Dermal; Dissociation; Dyes; Effector Cell; Environment; Flow Cytometry; Future; Genomics; Human; Human body; Image; Immune; Immune system; Immunocompromised Host; Individual; Infection; Infectious Skin Diseases; Inflammation; Investigation; Knowledge; Langerhans cell; Lesion; Life Cycle Stages; Mediating; Methodology; Methods; Microscope; Microscopy; Modeling; Molecular; Molluscum contagiosum virus; Monkeypox virus; Morbidity - disease rate; Morphology; Mus; Organ; Pathogenesis; Phase; Phenotype; Population; Poxviridae; Poxviridae Infections; Procedures; Production; Proteins; Proteomics; Protocols documentation; Recruitment Activity; Regulation; Resolution; Risk; Role; Sampling; Site; Skin; Smallpox; Staging; Staining method; Stains; Surface; System; Systemic infection; Systems Analysis; T-Lymphocyte; Tissues; Vaccines; Vaccinia virus; Vertebral column; Viral; Viral Vaccines; Virus; Virus Diseases; Virus Replication; Work; abstracting; adaptive immunity; base; cytokine; experience; imaging system; monocyte; mortality; novel; novel strategies; progenitor; programs; protein expression; response; transmission process; vector

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.

抽象的 病毒性皮肤感染是导致人类发病和死亡的一个重要原因，有数百万人 全世界感染痘病毒传染性软疣病毒 (MCV) 的个体，这种病毒仅存在于 由宿主免疫系统检查，因为免疫功能低下的个体出现全身衰弱的情况 感染。相关的痘病毒痘苗病毒（VACV）被用来为数亿人提供免疫 在天花根除计划期间，仍然是最广泛使用的病毒疫苗的支柱 向量。病毒感染期间，靶器官的形态发生显着变化，原因是 病毒生命周期进展过程中的细胞融合，以及多种反应物的涌入 免疫细胞。感染产生的炎症也可以刺激非经典细胞的分化。 驻留或浸润细胞群与通常存在于稳态的细胞不同。 因此，这些细胞的复杂表型分析需要使用大量的探针（大部分是 抗体）。然而，传统方法（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、定量和拓扑定位 细胞和蛋白质之间的相对关系，不会破坏和混淆组织解离，或分析 来自单个组织的 50-100 个参数。为了完成这项调查，我们将进行非侵入性的 使用 Toponome 成像系统（一种完全集成的成像循环仪显微镜）分析 50 多个参数 自动显微镜，允许大分子和细胞系统的空间分辨率显示 完整组织中存在数千种潜在的相互作用。我们将分析分子和细胞的变化 使用 VACV（痘病毒感染模型）感染小鼠皮肤后不同点的皮肤。 我们假设多重拓扑分析将揭示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， 在启动、控制和过程中驻留和浸润细胞群的定位和表型 VACV 皮肤感染的消退阶段。在目标 2 中，我们将检查两个单独的髓样细胞的作用 群体建立 VACV 病变的形态和浸润细胞的功能。我们 预计从这些研究中获得的结果将构成未来 RO1 或 PO1 提案的基础 感染细胞群、常驻细胞群和浸润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人体研究。