Spiga, Luisella (2016) Study of Salmonella pathogenicity mechanisms in vitro and in vivo. Doctoral Thesis.
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Salmonella enterica subsp. enterica includes many pathogenic serovars. Models of the study are S. typhimurium and S. abortusovis, a restricted serovar to sheep.
Bacterial pathogen’s transmission strategies are usually connected to pathogenesis and limited information is available about the immune response of sheep to S. abortusovis and about flagellin interaction with S. abortusovis. Knowledge about those factors are mainly based on studies of S. typhimurium.
Here we try to understand the role of flagella in S. abortusovis. The idea is that S. abortusovis highly regulates expression of flagella within the host, and we performed studies to clarify how S. abortusovis evade activation of the immune system.
Mutations in S. abortusovis flagella genes were generated and flagella from wild type and mutant’s strains of the two different serovars were extracted using diverse media to observe differences in flagellation and host interaction. Samples were analyzed by Western Blot to determine expression of major flagellar protein. Surface expression of flagella was verified by Flow Cytometry and by Scanning Electron Microscopy. To verify the capacity of S. typhimurium and S. abortusovis to stimulate TLR5, a colorectal
carcinoma cell line (T84 cells) was treated with purified flagellin and the transcriptional induction of a CXC chemokine IL-8 was measured.
We showed that S. abortusovis flagella are expressed in specific media, and they induced the transcription of IL-8 in T84 cells.
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