Ecotin protects Salmonella Typhimurium against the microbicidal activity of host proteases

Saposnik, L. M.; Coria, L. M.; Bruno, L.; Guaimas, F.; Pandolfi, J. B.; Pol, M.; Urga, M. E.; Sabbione, F.; McClelland, M.; Trevani, A.; Pasquevich, K. A.; Cassataro, J.

doi:10.1101/2024.05.15.594389

bioRxiv · 10.1101/2024.05.15.594389

Ecotin protects Salmonella Typhimurium against the microbicidal activity of host proteases

Saposnik, L. M.·Coria, L. M.·Bruno, L.·Guaimas, F.·Pandolfi, J. B.·Pol, M.·Urga, M. E.·Sabbione, F.·McClelland, M.·Trevani, A.·Pasquevich, K. A.·Cassataro, J.

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Abstract

Salmonella enterica serovar Typhimurium causes acute diarrhea upon oral infection in humans. The harsh and proteolytic environment found in the gastrointestinal tract is the first obstacle that these bacteria face after infection. However, the mechanisms that allow Salmonella to survive the hostile conditions of the gut are poorly understood. The ecotin gene is found in an extensive range of known phyla of bacteria and it encodes a protein that has been shown to inhibit serine proteases. Thus, in the present work we studied the role of ecotin of Salmonella Typhimurium in host-pathogen interactions. We found that Salmonella Typhimurium {Delta}ecotin strain exhibited lower inflammation in a murine model of Salmonella induced colitis. The {Delta}ecotin mutant was more susceptible to the action of pancreatin and purified pancreatic elastase. In addition, the lack of ecotin led to impaired adhesion to Caco-2 and HT-29 cell lines, related to the proteolytic activity of brush border enzymes. Besides, {Delta}ecotin showed higher susceptibility to lysosomal proteolytic content and intracellular replication defects in macrophages. In addition, we found Ecotin to have a crucial role in Salmonella against the microbicide action of granules released and neutrophil extracellular traps from human polymorphonuclear leukocytes. Thus, the work presented here highlights the importance of ecotin in Salmonella as countermeasures against the host proteolytic defense system. IMPORTANCEThe gastrointestinal tract is a very complex and harsh environment. Salmonella is a successful food borne pathogen, but little is known about its capacity to survive against the proteolysis of the gut lumen and intracellular proteases. Here, we show that Ecotin, a serine protease inhibitor, plays an important role in protecting Salmonella against proteases present at different sites encountered during oral infection. Our results indicate that Ecotin is an important virulence factor in Salmonella, adding another tool to the wide range of features this pathogen uses during oral infection.

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Saposnik, L. M., Coria, L. M., Bruno, L., Guaimas, F., Pandolfi, J. B., Pol, M., Urga, M. E., Sabbione, F., McClelland, M., Trevani, A., Pasquevich, K. A., Cassataro, J.. 2024-05-18. Ecotin protects Salmonella Typhimurium against the microbicidal activity of host proteases. https://doi.org/10.1101/2024.05.15.594389

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