Differential analysis of the genomic and transcriptomic mechanisms governing the intracellular life of atypical pathogenic Salmonella serovars in human and murine macrophages (360G-Wellcome-099837_Z_12_Z)

The aim of this project is to characterise and compare the ability of atypicalSalmonella serovars to survive and replicate following phagocytosis by both human and mouse macrophages. Salmonellosis is a zoonotic disease of major public health concern. Salmonella infections in humans have been mainly studied in the mouse model using Salmonella Typhimurium. However, the immunological response to Salmonella Typhimurium is completely different in humans to that of mice. Despite this, monocyte and macrophage studies have traditionally been carried out in murine cell lines (J774 and RAW). The interactions between a set of atypical Salmonella serovars and both human and mouse macrophages will be assessed at the bacterial and host level throughcell infection, cytokine detection/quantification and macrophage activation assays. SMRT sequencing of a specific serovar with an unusual infection phenotype will be performed to aid in genome assembly and to determine the methylome. dRNA-seq will be used to functionally annotate the genome and RNA-seq will be used to characterise the transcriptional landscape of the selected serovar. A TraDIS library will be generated for the specific Salmonella serovar to determine the essential gene set in in vitro and ex vivoconditions to identify the genes essential to survival in macrophages.