CCR Vaccine Branch FACS Core

CCR 疫苗分支 FACS 核心

• 批准号: 10703026
• 负责人: Katherine McKinnon
• 金额: $ 65.7万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10703026
• 关键词: Apoptosis; Biological; Blood specimen; Bromodeoxyuridine; CCR; COVID-19 vaccine; Cancer Biology; Cell Cycle; Cell Proliferation; Cells; Collaborations; Color; Containment; Core Facility; Cultured Cells; Event; Experimental Designs; Flow Cytometry; Fluorescent Dyes; Funding; Gene Expression; Goals; HIV; Human; Human T-lymphotropic virus 1; Human T-lymphotropic virus 2; Immunologic Monitoring; In Vitro; Infectious Agent; Label; Laboratories; Lasers; Monitor; Monoclonal Antibodies; Mus; NCI Center for Cancer Research; National Cancer Institute; National Institute of Allergy and Infectious Disease; Pediatric Oncology; Phenotype; Reagent; Receptor Biology and Gene Expression; Research Personnel; Research Project Grants; Retroviridae; Running; SIV; Safety; Sorting; Source; Speed; Stains; Techniques; Technology; Tissue Sample; Training; Vaccines; analog; biological research; cancer genetics; cellular oncology; cytokine; digital; experimental study; instrument; instrumentation; nonhuman primate; polyclonal antibody; release of sequestered calcium ion into cytoplasm; single cell analysis; vaccine trial

The Vaccine Branch FACS (Flow Cytometry) Core Facility has been created to meet an unmet need on the Bethesda Campus of NCI for a Flow Cytometry Core Facility with BL2/BL3 containment necessary to safely sort live cells infected with retroviruses, such as HIV/SIV, and other infectious agents known or suspected in blood and tissue samples including SARs. The facility has a multicolor cell sorter currently in use for standard and infectious sorting. In addition, this facility has analysis capabilities from five multicolor flow cytometry instruments that were donated to the facility from three Investigators in the CCR, and through IATAP funding. The facility purchased a 4 laser high end analyzer and a 3 laser high speed cell sorter. These were purchased at the beginning of FY09 and installed in Q2 FY09. The facility purchase a 28-color BD FACSymphony A5 in 2016. A new 28-color FACSymphony A5 is being purchase to replace a discontinued 12-color BD LSRII flow cytometer. The facility currently maintains one investigator-run 2 laser 4 color analog flow cytometer, two investigator-run 4 laser 18 color high end digital flow cytometers, two investigator run 5 laser 28 color high end digital flow cytometers.The facility also has two cell sorters: a 4 laser 16 color digital cell sorter equipped with containment for use as an infectious cell sorter and a 5 laser 19 color cell sorter in a biological safety cabinet. The facility currently supports more than 70 NCI/CCR Investigators from the Vaccine Branch, the Laboratory of Receptor Biology and Gene Expression, the Laboratory of Cellular Oncology, and the Laboratory of Cancer Biology and Genetics and Pediatric Oncology Branch. In addition to NCI investigators, the Facility Manager is working on a collaboration with an investigator from NEI and NIAID. Experiments currently conducted in the facility involve multi-parameter single cell analysis of cells from human, non-human primate and mouse primary sources. Additionally analysis is also conducted on in vitro cultured cells. Reagents for these experiments include monoclonal and polyclonal antibodies with fluorescent labels and fluorescent dyes used for staining a variety of cellular components. Current applications being supported by the facility include multi-color phenotypic analysis, cell cycle analysis, apoptosis analysis, proliferation analysis with BrdU and CFSE, intracellular cytokine analysis, rare event analysis and calcium flux analysis. Currently supported projects include, but are not limited to, immune monitoring for HIV, SIV, HTLV-1 and HTLV-2, and COVID-19 vaccine studies on non-human primates, and monitoring the results of gene expression on cell proliferation and apoptosis.

疫苗分部FACS（流式细胞术）核心设施的创建是为了满足NCI Bethesda校区对BL2/BL3遏制的流式细胞术核心设施的未满足需求，以安全分选感染逆转录病毒（如HIV/SIV）和其他已知或疑似血液和组织样本（包括SARs）中的感染因子的活细胞。该设施有一个多色细胞分选器，目前用于标准和传染性分选。此外，该设施还具备5台多色流式细胞仪的分析能力，这5台多色流式细胞仪是由CCR的3位研究人员和IATAP基金捐赠给该设施的。该设施购买了一台4激光高端分析仪和一台3激光高速细胞分选机。这些是在2009财年初购买的，并在2009财年第二季度安装。该设施于2016年购买了一台28色的BD FACSymphony A5。正在购买新的28色FACSymphony A5，以取代已停产的12色BD LSRII流式细胞仪。该设施目前拥有一台研究人员运行2激光4色模拟流式细胞仪，两台研究人员运行4激光18色高端数字流式细胞仪，两台研究人员运行5激光28色高端数字流式细胞仪。该设施还有两台细胞分选机：一台4激光16彩色数字细胞分选机配备了容器，用作感染性细胞分选机；另一台5激光19彩色细胞分选机安装在生物安全柜中。该设施目前支持来自疫苗部门、受体生物学和基因表达实验室、细胞肿瘤学实验室以及癌症生物学和遗传学实验室和儿科肿瘤学部门的70多名NCI/CCR调查员。除NCI调查员外，设施经理还与NEI和NIAID的一名调查员合作。目前在该设施进行的实验包括对来自人类、非人类灵长类动物和小鼠主要来源的细胞进行多参数单细胞分析。此外，还对体外培养的细胞进行了分析。用于这些实验的试剂包括带有荧光标记的单克隆和多克隆抗体和用于染色各种细胞成分的荧光染料。该设备目前支持的应用包括多色表型分析、细胞周期分析、细胞凋亡分析、BrdU和CFSE增殖分析、细胞内细胞因子分析、罕见事件分析和钙通量分析。目前支持的项目包括但不限于HIV、SIV、HTLV-1和HTLV-2的免疫监测，以及COVID-19疫苗在非人类灵长类动物中的研究，以及细胞增殖和凋亡基因表达的监测结果。