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The molecular mechanisms of virulence of the gastrointestinal pathogen  Salmonella enterica  are commonly studied using cell culture models of infection. In this work, we performed a direct comparison of the interaction of  S. enterica  serovar Typhimurium ( S.  Typhimurium) with the non-polarized epithelial cell line HeLa, the polarized cell lines CaCo2, T84 and MDCK, and macrophage-like RAW264.7 cells. The ability of  S.  Typhimurium wild-type and previously characterized auxotrophic mutant strains to enter host cells, survive and proliferate within mammalian cells and deploy the  Salmonella  Pathogenicity Island 2-encoded type III secretion system (SPI2-T3SS) was quantified. We found that the entry of  S.  Typhimurium into polarized cells was much more efficient than entry into non-polarized cells or phagocytic uptake. While SPI2-T3SS dependent intracellular proliferation was observed in HeLa and RAW cells, the intracellular replication in polarized cells was highly restricted and not affected by defective SPI2-T3SS. The contribution of aromatic amino acid metabolism and purine biosynthesis to intracellular proliferation was distinct in the various cell lines investigated. These observations indicate that the virulence phenotypes of  S.  Typhimurium are significantly affected by the cell culture model applied.

Divergent Roles of Salmonella Pathogenicity Island 2 and Metabolic Traits during Interaction of S. enterica Serovar Typhimurium with Host Cells