Transcriptional profiling of immune genes in bovine monocyte-derived macrophages exposed to bacterial antigens
Transcriptional profiling of immune genes in bovine monocyte-derived macrophages exposed to bacterial antigens
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DOI：
10.1016/j.vetimm.2010.12.002
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发表时间：
2011-03-15
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：
Taraktsoglou, Maria;Szalabska, Urszula;MacHugh, David E.
研究了 Toll 样受体 (TLR) 和其他免疫信号基因在用来自牛致病细菌的细菌产物攻击牛巨噬细胞期间的参与情况。建立了牛单核细胞来源的巨噬细胞（MDM）的体外细胞培养模型，并将这些细胞暴露于源自牛分枝杆菌的纯化蛋白衍生物（PPD-b）和源自大肠杆菌的脂多糖（LPS）。孵育24小时后，提取总RNA并通过实时定量逆转录PCR（qRT-PCR）测定免疫相关基因的表达。确定了跨越 TLR-2 和 TLR-4 途径的一系列基因的表达，从受体的最初激活到促炎细胞因子和趋化因子的产生。使用来自七只不同年龄匹配的奶牛的 MDM 进行的重复实验结果表明，PPD-b 处理导致 TLR2 和 TLR4 基因显着上调，并且 TLR 接头分子的表达谱表明这种信号传导是 MYD88 依赖性的。相反，LPS 通过不依赖 MYD88 的信号通路引起 TLR4 的显着上调。在 PPD-b 和 LPS 处理的样品中还检测到与 NF-κ B 信号传导相关的基因显着上调，并伴有促炎细胞因子（TNF、IL1B、IL6）和趋化因子基因（IL8、CCL5、CCL3）的表达。总体而言，LPS 攻击导致免疫相关基因更显着上调。此外，基因表达的倍数变化差异至少部分表明牛巨噬细胞由于LPS攻击而产生IFN-γ。 (C) 2010 Elsevier B.V. 保留所有权利。
The involvement of Toll-like receptors (TLRs) and other immune signalling genes during challenge of bovine macrophages with bacterial products derived from disease-causing bacteria in cattle was investigated. An in vitro cell culture model of bovine monocyte derived macrophages (MDM) was established and these cells were exposed to purified protein derivative (PPD-b) derived from Mycobacterium bovis and to lipopolysachharide (LPS) derived from Escherichia coli. Following 24 h incubation, total RNA was extracted and expression of immune related genes was determined by real time quantitative reverse transcription PCR (qRT-PCR). Expression of a selection of genes spanning the TLR-2 and TLR-4 pathways, from the initial activation of the receptors to the production of pro-inflammatory cytokines and chemokines was determined. Results from repeat experiments using MDM from seven different age-matched dairy cattle showed that PPD-b treatment caused significant up-regulation of the TLR2 and TLR4 genes and the expression profile of TLR adaptor molecules suggested that this signalling is MYD88-dependent. Conversely, LPS caused significant up-regulation of TLR4 via a MYD88-independent signalling pathway. Significant up-regulation of genes involved with NF-kappa B signalling was also detected in PPD-b- and LPS-treated samples accompanied by the expression of pro-inflammatory cytokine (TNF, IL1B, IL6) and chemokine genes (IL8, CCL5, CCL3). Overall, LPS challenge resulted in a more marked up-regulation of immune-related genes. Furthermore, the magnitude fold-change difference in gene expression suggests, at least in part, that bovine macrophages produce IFN-gamma, as a result of LPS challenge. (C) 2010 Elsevier B.V. All rights reserved.