Genetically Modifying Lactobacillus to Alter Gut Inflammation and Pathogens

这个项目是新的，过去一年的重点是表征表达 1型干扰素（IFN）由乳酸杆菌。开发了一种体外试验， 将表达小鼠β-干扰素基因的细胞直接加入小鼠巨噬细胞系中 并在孵育30分钟至1小时后测量STAT 1的磷酸化。我们 成功地证明了表达干扰素而非对照的细菌 细菌，触发了快速的STAT 1磷酸化，这种作用需要接触 细菌和巨噬细胞。此外，这种效果取决于 干扰素-β受体，但不需要Toll受体2或9。的表达 干扰素基因也诱导了原发性肝癌细胞中STAT 1的快速磷酸化。 鼠树突细胞以及小鼠肠上皮细胞系。此外，它似乎 通过小鼠树突状细胞改变白细胞介素-1 β（IL-1 β）的表达。电流 研究的重点是确定乳酸杆菌是否对葡聚糖钠有任何影响， 在小鼠模型系统中硫酸盐诱导的肠道炎症。

This project is new and this past year was focused on characterizing the expression of the Type 1 interferon (IFN) by Lactobacillus. An in vitro assay was developed whereby bacteria expressing the mouse interferon-beta gene were added directly to a mouse macrophage cell line and the phosphorylation of STAT1 was measured after 30 minutes to 1 hour of incubation. We successfully demonstrated that bacteria expressing the interferon but not the control bacteria, triggered rapid STAT1 phosphorylation and that this effect required contact between the bacteria and the macrophages. Furthermore, this effect was dependent on the interferon-beta receptor but did not require toll receptors 2 or 9. The expression of the interferon gene by the Lactobacillus also induced rapid STAT1 phosphorylation in primary murine dendritic cells as well as a mouse gut epithelial cell line. Furthermore, it appeared to alter the expression of interleukin-1 beta (IL-1beta) by murine dendritic cells. Current studies are focused on determining if the Lactobacillus have any effects on dextran sodium sulfate induced gut inflammation in mouse model systems.