College of Animal Science, Yangtze University, Jingzhou, 434025, Hubei, China.
Vet Res. 2018 Dec 12;49(1):121. doi: 10.1186/s13567-018-0616-9.
Peripheral blood transcriptome is an important intermediate data source for investigating the mechanism of Salmonella invasion, proliferation, and transmission. We challenged 4-week old piglets with Salmonella enterica serovar Typhimurium LT2 and investigated the peripheral blood gene expression profile before treatment (d0) and at 2 and 7 days post-inoculation (dpi) using deep sequencing. Regulator pathways were first predicted in silico and validated by wet-lab experiments. In total, 1255, 765, and 853 genes were differentially expressed between 2 dpi/d0, 7 dpi/d0, and 7 dpi/2 dpi, respectively. Additionally, 1333 genes showed a time effect during the investigated Salmonella infection period. Clustering analysis showed that the differentially expressed genes fell into six distinct expression clusters. Pathway annotation of these gene clusters showed that the innate immune system was first significantly upregulated at 2 dpi and then attenuated at 7 dpi. Toll-like receptor cascades, MyD88 cascade, phagosome pathway, cytokine signaling pathway, and lysosome pathway showed a similar expression pattern. Interestingly, we found that the ribosome pathway was significantly inhibited at 2 and 7 dpi. Gene expression regulation network enrichment analysis identified several candidate factors controlling the expression clusters. Further in vitro study showed that TGFB1 can inhibit Salmonella replication whereas TRP53 can promote Salmonella replication in porcine peripheral blood mononuclear cells and murine macrophages. These results provide new insights into the molecular mechanism of Salmonella-host interactions and clues for the genetic improvement of Salmonella infection resistance in pigs.
外周血转录组是研究沙门氏菌侵袭、增殖和传播机制的重要中间数据源。我们用鼠伤寒沙门氏菌 LT2 攻毒 4 周龄仔猪,在攻毒前(d0)和攻毒后 2 和 7 天(dpi)用深度测序技术检测外周血基因表达谱。首先通过计算机预测调控途径,然后通过湿实验验证。共鉴定到 1255、765 和 853 个基因在 2 dpi/d0、7 dpi/d0 和 7 dpi/2 dpi 之间差异表达。此外,1333 个基因在研究的沙门氏菌感染期间表现出时间效应。聚类分析显示差异表达基因分为六个不同的表达簇。这些基因簇的通路注释表明,固有免疫系统在 2 dpi 时首先显著上调,然后在 7 dpi 时减弱。Toll 样受体级联反应、MyD88 级联反应、吞噬体途径、细胞因子信号通路和溶酶体途径表现出相似的表达模式。有趣的是,我们发现核糖体途径在 2 和 7 dpi 时明显受到抑制。基因表达调控网络富集分析确定了几个控制表达簇的候选因子。进一步的体外研究表明,TGFB1 可以抑制沙门氏菌复制,而 TRP53 可以促进猪外周血单核细胞和鼠巨噬细胞中的沙门氏菌复制。这些结果为沙门氏菌-宿主相互作用的分子机制提供了新的见解,并为猪沙门氏菌感染抗性的遗传改良提供了线索。