CORE--GENE EXPRESSION PROFILING

免疫功能表现出衰老相关的缺陷，如T细胞对抗原的反应性降低。我们假设来自年轻和年老动物的等效细胞群之间的功能差异将表现为基因表达表型的改变。为了验证这一假设，我们将采用允许并行研究数千个转录本的DNA微阵列。由于解释数据所需的技术和分析工具的复杂性和成本，微阵列表达分析很难在单个实验室层面实施和支持。因此，我们将集中表达谱在一个核心基因表达谱。该核心将与OHSU的基因微阵列共享资源（GMSR）一起工作，从而利用这个完善的服务中心的现有资源。我们将对Lewinsohn、Nikolich-Zugich和Picker的项目进行基因表达谱分析，使用由GMSR打印的包含大约8900个不同克隆的人类cDNA阵列。阵列分析将包括从选定的细胞群体中分离RNA， RNA扩增，标记和杂交。数据定量和归一化将进行比较，从外周血单核细胞派生的无血缘关系的恒河猴普遍参考。微阵列结果的数据分析将由生物统计学和生物信息学核心提供支持，该核心是GMSR生物信息学单元的延伸。我们期望在老龄T细胞和抗原提呈细胞中检测到统计学上显著的转录变化。记录改变

Immunological functions display aging-associated defects such as decreased reactivity to antigen by T cells. We hypothesize that functional differences between equivalent cell populations from young and old animals will manifest themselves in altered gene expression phenotypes. To test this hypothesis we will employ DNA microarrays that allow to study thousands of transcripts in parallel. Due to the complexity and cost of both the technology and the analysis tools needed to interpret the data, microarray expression analysis is difficult to implement and support at the level of the individual laboratory. Therefore, we will centralize expression profiling in a core for gene expression profiling. This core will work together with the Gene Microarray Shared Resource (GMSR) of OHSU thus leveraging existing resources at this well established service center. We will perform gene expression profiling for projects by Lewinsohn, Nikolich-Zugich, and Picker using human cDNA arrays containing approximately 8900 different clones printed by the GMSR. Array analysis will encompass isolation of RNA from selected celt populations, RNA amplification, labeling and hybridization. Data quantitation and normalization will be performed in comparison to a universal reference derived from peripheral blood mononuclear cells of unrelated rhesus macaques. Data analysis of microarray results will be supported by the Biostatistics and Bionformatics Core which is an extension of the bioinformatics unit of the GMSR. We expect to detect statistically significant transcriptional changes in aged versus young T cells and antigen presenting cells. Transcripts changing most significantly will be selected for confirmation by quantitative PCR in an unrelated groups of rhesus macaques. The results are expected to identify profiles that reflect the aging process in these cells and might thus be linked mechanistically to the observed defects in immune responses.