Bacterial and host factors governing enterocyte exit of Salmonella

控制沙门氏菌肠上皮细胞排出的细菌和宿主因素

• 批准号: 446460159
• 负责人: Professor Dr. Mathias Walter Hornef
• 金额: --
• 依托单位: Institut für Medizinische Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/446460159?language=en
• 关键词: Bacterial; host; factors; governing; enterocyte

Hallmark of the pathogenesis of Salmonella is the ability to penetrate the intact intestinal epithelial barrier, reach the underlying lamina propria and evoke an inflammatory response. Penetration of the epithelial barrier requires invasion of non-phagocytic cells, intracellular survival, apical to basolateral transmigration and cell exit. Whereas cell invasion and intracellular survival have been extensively investigated during the last decades, the process the Salmonella transmigration and exit from epithelial cells has not obtained much attention. Based on our previous work, here we plan to functionally characterize the basolateral and apical enterocyte exit, screen for and analyze bacterial factors influencing enterocyte exit, and investigate apical enterocyte exit and its role in vivo in host transmission. To achieve these goals, life imaging and advanced electron microscopy, transposon mutagenesis and an iterative co-culture screen to identify genes promoting cell exit, targeted bacterial mutagenesis, and genetic fate mapping will be employed in combination with an in vitro co-culture model of polarized intestinal epithelial m-ICcl2 cells grown on transwell inserts, intestinal epithelial stem cell organoids and an established neonatal mouse infection model. The results are expected to significantly broaden our understanding of the intricate host-microbe interaction during the early course of the infection and may provide molecular targets for future preventive or therapeutic strategies.

沙门氏菌发病机制的标志是能够穿透完整的肠上皮屏障，到达下层固有层并引起炎症反应。上皮屏障的穿透需要非吞噬细胞的侵入、细胞内存活、顶侧至基底侧的迁移和细胞退出。尽管在过去的几十年中，细胞侵袭和细胞内存活已被广泛研究，但沙门氏菌从上皮细胞迁移和退出的过程并未得到太多关注。基于我们以前的工作，在这里，我们计划功能特性的基底外侧和顶端肠上皮细胞出口，筛选和分析影响肠上皮细胞出口的细菌因素，并调查顶端肠上皮细胞出口及其在体内宿主传播中的作用。为了实现这些目标，生命成像和先进的电子显微镜，转座子诱变和迭代共培养筛选，以识别促进细胞退出的基因，靶向细菌诱变和遗传命运作图将与transwell插入物上生长的极化肠上皮m-ICcl 2细胞的体外共培养模型，肠上皮干细胞类器官和建立的新生小鼠感染模型组合使用。这些结果有望显着拓宽我们对感染早期复杂的宿主-微生物相互作用的理解，并可能为未来的预防或治疗策略提供分子靶标。