Mutations in the Salmonella Typhi zzz genes impacts on OMVs internalization and uptake kinetics in epithelial cells in culture
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2020
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Universidad Andrés Bello
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Licencia CC
Resumen
Outer membrane vesicles (OMV) are nano-sized structures formed from the envelope of Gram-negative
bacteria (GNB). OMVs are involved in numerous processes are related to bacterial physiology and
pathogenesis. Furthermore, OMVs can also be used as biotechnological tools for delivering biomolecules
or as a platform to develop vaccines. In general, biotechnological applications are related to how the OMVs
interact with the eukaryotic cells. The Host – OMVs interaction depends on the internalization of OMVs by
cells. It has been proposed that the OMV size and protein cargo determine the internalization of OMVs by
non-phagocytic cells (i.e., epithelial cells). Recently, nine genes have been found to be involved in the OMV
biogenesis of Salmonella enterica serovar Typhi. These genes are generically called zzz genes and affect
both the OMV size and protein cargo. In the present work, we aimed to identify the differences that
mutations in the zzz genes may produce in the Host cells – OMVs interaction. We first evaluated the
capacity of the OMVs from the ∆zzz mutants to be internalized by the epithelial cell line HT-29. We
observed that all the OMVs could be internalized, and its internalization increased with time. Then we
assessed the difference in Host cells – OMVs interaction that the OMVs from the ∆zzz mutants may have
between them and the WT strain. We observed a difference in the mean fluorescence that the OMVs
exhibited inside the epithelial cells, including the marking pattern inside the cells, the uptake kinetics, and
entry in two temperatures (37 °C and 4 °C) for OMV internalization. We also showed that mutations in the
zzz genes directly impact the host cells – OMVs interaction. Thus, these results allowed us to better
understand how S Typhi OMVs interact with host cells. We expect this knowledge to set the foundations
of future developments of a new biotechnological tool based on OMVs technology as well as a better
understanding of the impact that OMV have during S. Typhi infection.
Notas
Tesis (Magíster en Biotecnología y Ciencias de la Vida)
Palabras clave
Salmonella Typhi, Investigaciones