Salmonella enterica species are intracellular bacteria causative agents of gastroenteritis and typhoid fever in humans. Pregnancy poses an increased risk of severe Salmonellosis in many mammalian species contributing to miscarriage and/or maternal illness. Previous studies indicated that Salmonella infection in pregnant mice caused rapid fetal and maternal death due to massive bacterial proliferation in the placenta. However, the susceptibility of human primary trophoblast cells (cTBCs) to Salmonella infection was not known. We hypothesized that human placental trophoblast cells are productively infected and provide a unique intracellular niche that permits uncontrolled Salmonella replication due to an ineffective maternal innate immune response to the virulent bacteria resulting in placental death. Firstly, we observed that S.Tm strains defective in the Salmonella pathogenicity island (SPI)-1 type III secretion system (TTSS) (S.Tm-ΔinvA) were unable to enter epithelial cells, but efficiently infected placental choriocarcinoma cell lines through scavenger receptor-mediated endocytosis. Next, we observed that S.Tm failed to grow vigorously in macrophages, but replicated rapidly within epithelial and placental trophoblast cells. Further examination of intracellular localization of S.Tm indicated that bacteria were arrested in early Rab5 expressing phagosomal vesicles within trophoblast cells, whereas phagosomal maturation progressed steadily in macrophages (with expression of lysosomal-associated membrane protein-1 (LAMP-1) and cathepsin D). Moreover, human primary cTBCs harboring S.Tm underwent rapid death of the cells. Infected cTBCs expressed phosphorylated-receptor-interacting serine/threonine-protein kinase (RIPK)-1 protein and phosphorylated-mixed lineage kinase domain-like (MLKL), suggesting induction of the necroptosis pathway of cell death. Furthermore, specific inhibition of necroptosis rescued S.Tm-induced death of cTBCs. Finally, S.Tm infected trophoblast cells produced interleukin (IL)-10, and signal transducer and activator of transcription (STAT)-3 signalling. This correlated to delayed phagosomal maturation which consequently facilitated intracellular pathogen proliferation. Overall, human trophoblast cells may act as reservoirs for S.Tm survival and may aid dissemination in the pregnant host.
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/36032 |
| Date | January 2017 |
| Creators | Nguyen, Tina |
| Contributors | Krishnan, Lakshmi |
| Publisher | Université d'Ottawa / University of Ottawa |
| Source Sets | Université d’Ottawa |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
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