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Antigen stimulation of peripheral blood mononuclear cells from Mycobacterium bovis infected cattle yields evidence for a novel gene expression program

机译:牛牛分枝杆菌感染牛外周血单核细胞的抗原刺激产生了新的基因表达程序的证据

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摘要

BackgroundBovine tuberculosis (BTB) caused by Mycobacterium bovis continues to cause substantial losses to global agriculture and has significant repercussions for human health. The advent of high throughput genomics has facilitated large scale gene expression analyses that present a novel opportunity for revealing the molecular mechanisms underlying mycobacterial infection. Using this approach, we have previously shown that innate immune genes in peripheral blood mononuclear cells (PBMC) from BTB-infected animals are repressed in vivo in the absence of exogenous antigen stimulation. In the present study, we hypothesized that the PBMC from BTB-infected cattle would display a distinct gene expression program resulting from exposure to M. bovis. A functional genomics approach was used to examine the immune response of BTB-infected (n = 6) and healthy control (n = 6) cattle to stimulation with bovine tuberculin (purified protein derivative – PPD-b) in vitro. PBMC were harvested before, and at 3 h and 12 h post in vitro stimulation with bovine tuberculin. Gene expression changes were catalogued within each group using a reference hybridization design and a targeted immunospecific cDNA microarray platform (BOTL-5) with 4,800 spot features representing 1,391 genes.
机译:背景技术由牛分枝杆菌引起的牛结核病(BTB)继续给全球农业造成重大损失,并对人类健康产生重大影响。高通量基因组学的出现促进了大规模的基因表达分析,为揭示分枝杆菌感染的分子机制提供了新的机会。使用这种方法,我们以前已经证明,在没有外源抗原刺激的情况下,来自BTB感染动物的外周血单核细胞(PBMC)中的先天免疫基因在体内受到抑制。在本研究中,我们假设来自BTB感染牛的PBMC将显示出由于暴露于牛分枝杆菌而产生的独特基因表达程序。功能基因组学方法用于检查BTB感染(n = 6)和健康对照(n = 6)牛对牛结核菌素(纯化的蛋白衍生物– PPD-b)刺激的免疫应答。在用牛结核菌素体外刺激之前和之后3小时和12小时收获PBMC。使用参考杂交设计和靶向免疫特异性cDNA微阵列平台(BOTL-5),将每组中的基因表达变化进行分类,并具有代表1,391个基因的4,800个斑点特征。

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