Genetically Modifying Lactobacillus to Alter Gut Inflammation and Pathogens

这个项目是新的，在过去的一年里，重点是表征乳杆菌表达的1型干扰素(干扰素)。建立了一种体外实验，将表达小鼠干扰素-β基因的细菌直接加入到小鼠巨噬细胞系中，并在孵育30分钟至1小时后检测STAT1的磷酸化。我们成功地证明了表达干扰素的细菌而不是对照细菌触发了STAT1的快速磷酸化，这一效应需要细菌与巨噬细胞之间的接触。此外，这种作用依赖于干扰素-β受体，而不需要Toll受体2或9。乳杆菌表达干扰素基因还诱导了原代小鼠树突状细胞和小鼠肠道上皮细胞株中STAT1的快速磷酸化。此外，它似乎改变了小鼠树突状细胞的白介素1β(IL-1β)的表达。目前的研究集中在确定乳酸菌对葡聚糖硫酸钠诱导的小鼠模型系统的肠道炎症是否有任何影响。

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.