Gene expression profiling of peripheral blood mononuclear cells (PBMC) from Mycobacterium bovis infected cattle after in vitro antigenic stimulation with purified protein derivative of tuberculin (PPD)
Gene expression profiling of peripheral blood mononuclear cells (PBMC) from Mycobacterium bovis infected cattle after in vitro antigenic stimulation with purified protein derivative of tuberculin (PPD)
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DOI：
10.1016/j.vetimm.2006.04.012
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发表时间：
2006-09-15
影响因子：
1.8
通讯作者：
MacHugh, David E.
中科院分区：
文献类型：
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作者：
Meade, Kieran G.;Gormley, Eamonn;MacHugh, David E.
使用信使 RNA (mRNA) 丰度的微阵列分析来研究感染牛分枝杆菌（牛结核病病原体）的牛的外周血单核细胞 (PBMC) 的基因表达程序。使用具有代表 1336 个基因的斑点特征的免疫特异性牛微阵列平台 (BOTL-4)，对来自用牛纯化的结核菌素蛋白衍生物 (PPD-牛) 体外刺激的六只牛支原体感染的牛的 PBMC 进行转录分析。在四个时间点（刺激后 3 小时、6 小时、12 小时和 24 小时）收获细胞，并使用混合样品的裂区设计进行微阵列实验，以比较每个时间点 PPD 牛刺激的 PBMC 和未刺激对照之间的基因表达。对这些数据的统计分析显示，在 24 It 时间过程中，刺激和未刺激的 PBMC 之间有 224 个基因（类似于阵列上转录本的 17%）存在差异表达（P < 0.05）。在 24 小时内用 PPD 牛刺激的牛支原体感染的 PBMC 中，在 224 个基因中，87 个基因显着上调，137 个基因显着下调。然而，PBMC 转录组的扰动在刺激后 3 小时和 12 小时的时间点最为明显，分别有 81 和 84 个基因差异表达。此外，更严格的统计阈值（P < 0.01）揭示了 35 个基因（接近 3%）在整个时间过程中存在差异表达。
Microarray analysis of messenger RNA (mRNA) abundance was used to investigate the gene expression program of peripheral blood mononuclear cells (PBMC) from cattle infected with Mycobacterium bovis, the causative agent of bovine tuberculosis. An immunospecific bovine microarray platform (BOTL-4) with spot features representing 1336 genes was used for transcriptional profiling of PBMC from six M. bovis-infected cattle stimulated in vitro with bovine purified protein derivative of tuberculin (PPD-bovine). Cells were harvested at four time points (3 h, 6 h, 12 It and 24 h post-stimulation) and a split-plot design with pooled samples was used for the microarray experiment to compare gene expression between PPD-bovine stimulated PBMC and unstimulated controls for each time point. Statistical analyses of these data revealed 224 genes (similar to 17% of transcripts on the array) differentially expressed between stimulated and unstimulated PBMC across the 24 It time course (P < 0.05). Of the 224 genes, 87 genes were significantly upregulated and 137 genes were significantly downregulated in M. bovis-infected PBMC stimulated with PPD-bovine across the 24 h time course. However, perturbation of the PBMC transcriptome was most apparent at time points 3 It and 12 h post-stimulation, with 81 and 84 genes differentially expressed, respectively. In addition, a more stringent statistical threshold (P < 0.01) revealed 35 genes (similar to 3%) that were differentially expressed across the time course.