Project description:Many non-typhoidal serovars of Salmonella such as Salmonella enterica serovar Typhimurium (S. Typhimurium) are the leading cause of food-borne gastroenteritis, resulting in millions of infections each year and sometimes death. Salmonella enterica serovar Typhimurium is the most common non-typhoidal Salmonella strain isolated from patients around the world and is used as a mouse model to study bacterial pathogenesis and host-microbe interactions. Furthermore, S. Typhimurium is an important pathogen in livestock animals including chickens and cattle. S. Typhimurium utilises a multitude of virulence factors to reach and invade host cells and for its intracellular survival. However, little is known about the mechanism of protein synthesis of these virulence factors at the codon level. Here, we performed RNA-seq and ribosome profiling. Ribosome profiling allows the global mapping of translating ribosomes on the transcriptome and therefore provides direct measure of protein synthesis.
Project description:1-day-old C57BL/6 mice were left untreated (co) or orally infected with 10E2 CFU wildtype (wt) or delta invC SPI1 mutant Salmonella Typhimurium (ATCC14028). Four biological replicates obtained from individual animals were exmained; each group contained animals from at least 2 different litters. On day 4 p.i., animals were sacrificed and intestinal epithelial cells were isolated from total small intestine (protocol according to: Lotz et al., J. Exp. Med. 2006). Total RNA was isolated by TriZol and its purity was examined using a Bioanalyzer. We used microarrays to detail the global gene expression in primary total isolated intestinal epithelial cells.
Project description:Salmonella enterica causes serious global burden of morbidity and mortality and is a major cause of infant bacteremia in sub Saharan Africa. Diseases caused by Salmonella are treatable with antibiotics but successful antibiotic treatment has become difficult due to antimicrobial resistance. An effective vaccine together with public health effort may therefore be a better strategy to control these infections. Protective immunity against Salmonella depends primarily on T cell-mediated immune responses and therefore identifying relevant T cell antigens is necessary for Salmonella vaccine development. Our laboratory has used an immunoproteomics approach to identify Chlamydia T cell antigens that exhibited significant protection against Chlamydia infection in mice. In this study, we infected murine bone marrow derived dendritic cells from C57BL/6 mice with Salmonella enterica strain SL1344 followed by isolation of MHC class I and II- molecules and elution of bound peptides. The sequences of the peptides were then identified using tandem mass spectrometry. We identified 87 MHC class II and 23 MHC class I Salmonella derived peptides. Four of 12 peptides stimulated IFN-? production by CD4 T cells from the spleens of mice with persistent Salmonella infection. These antigens will be useful for Salmonella immunobiology research and are potential Salmonella vaccine candidates.