The Cell Phenotyping and Mouse Core

项目摘要 细胞表型和小鼠核心，指定为核心C，是一个集中的设施，将提供 研究者用一个完善的和可重复的甲型流感诱导的肺炎模型。核心C将 产生流感（A/WSN/33）病毒的储备，滴定和维持病毒储备，并进行 项目1、2、3和4中概述的实验动物。定量工具，如排放测量 肺力学，定量组织学评估，流式细胞术表型和分类的 免疫细胞群和促炎细胞因子的多重测量将由Core C根据项目1、2、3和4中概述的研究计划。核心C将使用先进的流式细胞术 定量、表型和分选特定炎症和实质细胞群（例如， 组织驻留和募集的肺泡巨噬细胞，血管周围和支气管周围间质巨噬细胞， 单核细胞亚群，树突状细胞亚群，T、B和NK细胞亚群，肺泡上皮II型细胞，成纤维细胞 亚群，内皮细胞）。的排序和隔离功能 核心将允许通过RNA测序对转录组反应进行细胞类型特异性评估，包括 单细胞RNA测序。为此，核心将提供自动DNA/RNA分离从分选细胞， 随后进行质量评估和RNA测序文库构建和测序。核心C将繁殖 小鼠产生细胞类型或组织特异性Cre重组酶系以诱导组织特异性敲除 小鼠核心C将对项目负责人提出的所有鼠品系进行基因分型。核心C将 维持野生型和基因工程小鼠将在其中维持的统一环境， 使其遭受肺损伤并允许其恢复。这些工具共同为Project提供了独特的资源 研究者，如果没有PPG的支持，很难复制。

Project Summary The Cell Phenotyping and Mouse Core, designated as Core C, is a centralized facility that will provide investigators with a well-established and reproducible model of influenza A–induced pneumonia. Core C will generate stocks of influenza (A/WSN/33) virus, titrate and maintain viral stocks, and perform infection of the experimental animals as outlined in Projects 1, 2, 3 and 4. Quantitative tools such as Flexivent measurements of lung mechanics, quantitative histological assessment, flow cytometric phenotyping and sorting of the immune cellular populations, and multiplex measures of pro-inflammatory cytokines will be conducted by Core C according to the research plan outlined in Projects 1, 2, 3 and 4. Core C will use advanced flow cytometry capabilities to quantify, phenotype, and sort specific inflammatory and parenchymal cell populations (such as tissue-resident and recruited alveolar macrophages, perivascular and peribronchial interstitial macrophages, monocyte subsets, dendritic cell subsets, T, B and NK cell subsets, alveolar epithelial type II cells, fibroblast subsets, endothelial cells) from the murine lung and lymphoid organs. The sorting and isolation capabilities of the Core will allow cell-type-specific assessment of transcriptomic response via RNA-sequencing, including single-cell RNA-sequencing. To this matter, Core will provide automated DNA/RNA isolation from sorted cells, followed by quality assessment and RNA-sequencing library construction and sequencing. Core C will breed mice to generate the cell-type- or tissue-specific Cre recombinase lines to induce tissue-specific knockout mice. Core C will perform the genotyping of all the murine strains proposed by Project Leaders. Core C will maintain a uniform environment in which wild-type and genetically engineered mice will be maintained, subjected to lung injury and allowed to recover. Collectively, these tools provide a unique resource for Project Investigators, which would be difficult to reproduce without the support of this PPG.