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Quantitative analysis of the immune response upon Salmonella typhimurium infection along the porcine intestinal gut

机译:猪肠道内鼠伤寒沙门氏菌感染后免疫反应的定量分析

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摘要

Salmonella enterica serovar Typhimurium causes enteric disease and compromises food safety. In pigs, the molecular response of the intestine to S. typhimurium has been traditionally characterized by in vitro models that do not reflect the actual immunological competence of the intestinal mucosa. In this work, we performed an oral S. typhimurium infection study to obtain insight into the in vitro response in three different sections (jejunum, ileum and colon) of the porcine intestine. For this, samples from one-month-old infected piglets were collected during a time course comprising 1, 2, and 6 days post inoculation to evaluate the intestinal response by quantifying the mRNA expression of gene coding for 28 innate immune system molecules using quantitative real-time PCR assays. In addition, samples from non-infected control animals were also employed to establish differences in the steady state gene expression between intestinal sections. The panel of quantified molecules included an assortment of cytokines, chemokines, pattern-recognition receptors, intracellular signaling molecules, transcription factors and antimicrobial molecules. Changes in gene expression occurred in the three different parts of the intestine and during the course of the S. typhimurium infection. Moreover, the high variation observed in expression patterns of genes coding for inflammatory mediators could indicate that each intestinal section responds differently to the infection. Thus, on the contrary to findings in the jejunum and colon, a down-regulation and lack of induction of some proinflammatory cytokine transcripts was observed in the ileum. Nevertheless, all chemoattractant cytokines assayed were up-regulated in the ileum and jejunum whereas only interleukin-8 and MIP-1α mRNA were over expressed in the colon. In conclusion, our results reveal regional differences in gene expression profiles along the porcine intestinal gut as well as regional differences in the inflammatory response to S. typhimurium infection. Taken together, these data should provide a basis for a complete understanding of the porcine intestinal response to bacterial infection.
机译:肠炎沙门氏菌鼠伤寒沙门氏菌会引起肠道疾病并危害食品安全。在猪中,肠对鼠伤寒沙门氏菌的分子反应传统上是通过体外模型来表征的,该模型不能反映肠粘膜的实际免疫能力。在这项工作中,我们进行了口服鼠伤寒沙门氏菌感染研究,以深入了解猪肠三个不同部分(空肠,回肠和结肠)的体外反应。为此,在接种后1、2和6天的时间过程中收集了一个月大的受感染仔猪的样品,以定量定量编码28种先天免疫系统分子的基因的mRNA表达来评估肠道反应PCR检测。另外,还使用来自未感染对照动物的样品来建立肠段之间稳态基因表达的差异。一组定量的分子包括各种细胞因子,趋化因子,模式识别受体,细胞内信号分子,转录因子和抗微生物分子。基因表达的变化发生在肠的三个不同部分以及鼠伤寒沙门氏菌感染过程中。此外,在编码炎症介质的基因表达模式中观察到的高度变异可能表明每个肠段对感染的反应不同。因此,与在空肠和结肠中发现的相反,在回肠中观察到一些促炎性细胞因子转录物的下调和缺乏诱导。尽管如此,所有测定的趋化性细胞因子在回肠和空肠中均被上调,而结肠中仅白介素8和MIP-1αmRNA过度表达。总之,我们的结果揭示了沿猪肠道的基因表达谱的区域差异以及对鼠伤寒沙门氏菌感染的炎症反应的区域差异。综上所述,这些数据应为全面了解猪对细菌感染的肠道反应提供基础。

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