Contribution of typhoid toxin in the pathogenesis of Salmonella Typhi

Abstract

To persist and establish infection, Salmonella utilizes a battery of different virulence determinants at every stage of infection. Typhoid toxin, a newly identified toxin in Salmonella enterica serovar Typhi is recognized as one of the virulence factors that has been linked with Salmonella pathogenesis. In this study, we have further investigated the role of typhoid toxin in the symptomatology of typhoid fever through in-vivo and ex-vivo studies. In mice, administration of cloned and purified typhoid toxin induces similar symptoms observed during typhoid fever such as fever, weight loss with a decrease in peripheral leucocyte count along with an increase in levels of pro-inflammatory cytokines (Il-6, TNF-α). Results of DNA analysis, fluorescence microscopy and flow cytometry of typhoid toxin-treated macrophages (ex-vivo) altogether revealed the CdtB (subunit of typhoid toxin) mediated DNA damage that led to the apoptosis of cells. Furthermore, to validate CdtB's catalytic role, macrophages were treated with typhoid toxin preincubated with anti-CdtB antibodies (generated in mice). Re-assessment of macrophage DNA by gel electrophoresis and flow cytometry analysis indicated a significant decrease in DNA damage and cells undergoing apoptosis, respectively. Moreover, a significant reduction in in-vitro DNase activity of CdtB protein was also observed on preincubating holotoxin with anti-CdtB antibodies. In total, this study highlights the role of typhoid toxin in inducing typhoid fever-like symptomatology, which may be executed through the toxin's catalytic subunit CdtB.