Core--Laboratory

核心--实验室

• 批准号: 7321672
• 负责人: M CHRISTINE ZINK
• 金额: $ 20.13万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7321672
• 关键词: Aliquot; Archives; Astrocytes; Autopsy; Biological Assay; Blood; Bone Marrow; Brain; CD8-Positive T-Lymphocytes; Cell Line; Cells; DNA; Databases; HIV; Laboratories; Lymph Node Tissue; Lymphocyte; Macaca; Microglia; Peripheral Blood Lymphocyte; Plasma; Plasma Cells; Polymerase Chain Reaction; Preparation; Primary Cell Cultures; Process; RNA; RNA-Directed DNA Polymerase; Reagent; SIV; Sampling; Spleen; Time; Tissues; Viral Load result; Virus; lymph nodes; macrophage; monocyte; p27 Cell Cycle Protein; p27 Enzyme Inhibitor; programs; research study

The SIV Biosafety Level 3 (BL3) Core Laboratory will be directed by Dr. M. Christine Zink. The BL3 technical staff will be responsible for processing all samples from the infected and uninfected control macaques throughout the experiments described in this Program and will provide samples and reagents to all projects in the program. This core laboratory will fulfill the following Specific Aims: To provide expert support in the preparation of SIV and HIV stocks, and quantitation of virus stocks for all projects. To receive, process, aliquot and log into appropriate databases all samples from infected and uninfected macaques. To isolate lymphocytes from blood (and lymph nodes, spleen and brain at necropsy) of uninfected and infected macaques to quantitate and perform longitudinal characterization of the activation and effector state of CD4+/CD8 + and cells and to use these cells in experiments in Projects 1 and 3. To isolate monocytes from blood of uninfected and infected macaques for experimental studies in Projects 1-4. To isolate macrophages/microglia and astrocytes from the brains of all infected and uninfected macaques for analysis for Projects 1, 2 and 4. To aliquot and archive plasma and CSF samples for viral load and other assays. To prepare samples for Real Time PCR, extraction of RNA or DNA for Projects 1 and 2. To process, aliquot and archive blood cells and plasma, CSF and other tissues at necropsy of each macaque in the study as described in the specific projects. To establish primary cell cultures, and maintain cell lines.

SIV生物安全3级（BL3）核心实验室将由M. Christine Zink博士执导。 BL3技术人员将负责在本计划中描述的整个实验中处理来自受感染和未感染的控制猕猴的所有样品，并将为该计划中的所有项目提供样品和试剂。 该核心实验室将实现以下具体目标：在准备SIV和HIV股票方面提供专家支持，并为所有项目量化病毒股票。要接收，处理，等分并登录适当的数据库中的所有样品，这些样本都来自感染和未感染的猕猴。从未感染和感染的猕猴中分离出血液（和淋巴结，脾脏和大脑）的血液（和淋巴结，脾脏和大脑），以定量和执行CD4 +/CD8 +的激活和效应状态的纵向表征，并在项目1和3的实验中使用这些细胞的cd4 +/cd8 +的效应状态，以分离型和3。的细胞中，以分离型和3。 1-4。分离巨噬细胞/小胶质细胞和星形胶质细胞 所有受感染和未感染的猕猴的大脑，用于分析项目1、2和4。到等分等分等离子体和CSF样品，以进行病毒载荷和其他测定。为了准备实时PCR样品，将RNA或DNA提取为项目1和2。在研究中，在研究中所述的每个猕猴的尸检中，等分试样和档案血细胞以及等离子体，CSF和其他组织。建立原代细胞培养并维持细胞系。