Core--Immunology

核心--免疫学

• 批准号: 6354593
• 负责人: Malek Kamoun
• 金额: $ 23.33万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6354593
• 关键词: biomedical facility; cytokine; enzyme linked immunosorbent assay; flow cytometry; immunology

The Immunology Core will perform multi-parameter flow cytometry assays and cytokine assays. Dr. Kamoun, will be the Director of this core. He is the Director of the Clinical Immunology Laboratory at the University of Pennsylvania Medical Center. The Clinical Immunology Laboratory provides a wide range of laboratory tests on patients with autoimmune diseases including MS, and SLE as well as advanced multi- parameter four-color flow cytometry-based assays. Alterations in subset of CD4+ and CD8+ T cells and B cells can be determined in MS and SLE patients treated with antibody therapies. Lymphocyte activation can be evaluated assessing expression CD4+, CD8+ or B cells of a series of activation markers and adhesion molecules. Analysis of constitutive and induced cytokine production by intracellular staining of permeabilized cells can be performed using whole blood single cell cytokine assays which was recently established in our laboratory. This method allows the identification of cells producing cytokine while simultaneously analyzing the surface phenotype. In vitro production of cytokine can also be measured using ELISA-based assays. We can measured IL-2, IL-4, IL-10, IL-12 (p40 and the biologically active p70 heterodimer), INF-gamma, TNF-alpha and LT-alpha. All reagents used in the Laboratory are subjected to quality control procedures prior to use. All results of QC procedures are reviewed by the supervisor with corrective action implemented as necessary. Our lab participates in QA programs including the CAP surveys quarterly and in the CDC T cell immunophenotyping performance evaluation program. Performance of the flow cytometers is monitored daily using a suspension of beads to demonstrate scatter, fluorescence, and compensation parameters within defined limits. The Technologists employed in this section of the laboratory are each qualified under CLIA88 guidelines to perform "high complexity testing". The Laboratory Director and the Supervisor meet CLIA 88 requirements as director and technical supervisor respectively. The laboratory maintains a comprehensive safety program for all employees encompassing OSHA regulation and conforming to all Federal, State and local regulation.

免疫学核心将执行多参数流式细胞仪测定法和细胞因子测定。 Kamoun博士将担任该核心的主任。他是宾夕法尼亚大学医学中心的临床免疫学实验室主任。临床免疫学实验室为包括MS和SLE的自身免疫性疾病以及高级多参数四色流式细胞仪测定法提供了广泛的实验室测试。可以在MS和用抗体疗法治疗的SLE患者中确定CD4+和CD8+ T细胞和B细胞子集的改变。可以评估淋巴细胞激活评估一系列激活标记和粘附分子的表达CD4+，CD8+或B细胞。通过全血单细胞因子分析，可以对透化细胞的细胞内染色进行组成型和诱导的细胞因子产生分析，该分析最近在我们的实验室中建立。该方法允许鉴定产生细胞因子的细胞，同时分析表面表型。还可以使用基于ELISA的测定法测量细胞因子的体外产生。我们可以测量IL-2，IL-4，IL-10，IL-12（P40和生物活性P70异二聚体），INF-Gamma，TNF-Alpha和LT-Alpha。在使用前，实验室中使用的所有试剂均经过质量控制程序。质量控制程序的所有结果均由主管审查，并根据需要采取纠正措施。我们的实验室参与质量检查计划，包括季度CAP调查和CDC T细胞免疫表型性能评估计划。每天使用珠子的悬浮液来监测流式细胞仪的性能，以证明定义限制内的散射，荧光和补偿参数。实验室这一部分中使用的技术人员根据CLIA88指南进行了“高复杂性测试”的资格。实验室主任和主管分别符合CLIA 88的要求，分别担任董事和技术主管。该实验室为所有涵盖OSHA法规的员工提供了一项综合安全计划，并符合所有联邦，州和地方法规。