Unlike most salmonellae, Salmonella enterica serovar Typhi causes life-threatening systemic infections known as typhoid fever, for which the molecular basis is unknown; here administration of typhoid toxin produced by S. Typhi reproduces many of the acute symptoms of typhoid fever, carbohydrates on cell surface glycoproteins are identified as receptors for typhoid toxin and the toxin’s crystal structure is determined, providing insights into these interactions. The biological basis for the pathogenic properties of Salmonella enterica serovar Typhi (S. Typhi) is largely unknown. S. Typhi causes life-threatening systemic infections known as typhoid fever, whereas most other Salmonella enterica serotypes are either harmless or associated with less-serious gastric infections or food poisoning. This study shows that administration of typhoid toxin, a novel AB toxin composed of two A subunits unique to S. Typhi, reproduces many of the acute symptoms of typhoid fever. The authors go on to identify carbohydrates on cell-surface glycoproteins as receptors for typhoid toxin and determine the toxin's crystal structure, providing insights into these interactions. Theses advances suggest that antitoxin-based therapeutics may be effective in treating typhoid. Salmonella enterica serovar Typhi (S. Typhi) differs from most other salmonellae in that it causes a life-threatening systemic infection known as typhoid fever1. The molecular bases for its unique clinical presentation are unknown2. Here we find that the systemic administration of typhoid toxin, a unique virulence factor of S. Typhi, reproduces many of the acute symptoms of typhoid fever in an animal model. We identify specific carbohydrate moieties on specific surface glycoproteins that serve as receptors for typhoid toxin, which explains its broad cell target specificity. We present the atomic structure of typhoid toxin, which shows an unprecedented A2B5 organization with two covalently linked A subunits non-covalently associated to a pentameric B subunit. The structure provides insight into the toxin’s receptor-binding specificity and delivery mechanisms and reveals how the activities of two powerful toxins have been co-opted into a single, unique toxin that can induce many of the symptoms characteristic of typhoid fever. These findings may lead to the development of potentially life-saving therapeutics against typhoid fever.

The vigorous cytokine response of immune cells to Gram-negative bacteria is primarily mediated by a recognition molecule, Toll-like receptor 4 (TLR4), which recognizes lipopolysaccharide (LPS) and initiates a series of intracellular NF-κB–associated signaling events. Recently, bladder epithelial cells (BECs) were reported to express TLR4 and to evoke a vigorous cytokine response upon exposure to LPS. We examined intracellular signaling events in human BECs leading to the production of IL-6, a major urinary cytokine, following activation by Escherichia coli and isolated LPS. We observed that in addition to the classical NF-κB–associated pathway, TLR4 triggers a distinct and more rapid signaling response involving, sequentially, Ca²⁺, adenylyl cyclase 3–generated cAMP, and a transcriptional factor, cAMP response element–binding protein. This capacity of BECs to mobilize secondary messengers and evoke a more rapid IL-6 response might be critical in their role as first responders to microbial challenge in the urinary tract. In spite of frequent cross contamination by bacteria from the gut, urinary tract infections are relatively infrequent. Although much of the credit goes to cells lining the urinary tract, such as bladder cells, how this is achieved remains unclear. Human bladder cells display, on their surfaces, special molecules called Toll-like receptors, which sense the presence of bacteria and trigger the cells to release a variety of chemicals called cytokines. Cytokines contribute to the recruitment of phagocytic cells from the blood to the site of infection to clear bacteria. In this paper, we reveal that the Toll-like receptor–initiated intracellular signals leading to the production of cytokines by bladder cells involve the same pathway seen in other cells, as well as an additional and more rapid signaling pathway. Rapid production of cytokines by bladder cells will facilitate early clearance of bacteria. Additionally, possession of multiple signaling pathways by bladder cells for producing cytokines is advantageous because bacteria that infect the urinary tract have the capability to suppress certain signaling events that lead to cytokine production by bladder cells.

UropathogenicEscherichia coliinvade bladder epithelial cells (BECs) by direct entry into specialized cAMP regulated exocytic compartments. Remarkably, a significant number of these intracellular bacteria are subsequently expelled in a nonlytic and piecemeal fashion by infected BECs. Here, we report that expulsion of intracellularE. coliby infected BECs is initiated by the pattern recognition receptor, Toll-like receptor (TLR)4, after activation by LPS. Also, we reveal that caveolin-1, Rab27b, PKA, and MyRIP are components of the exocytic compartment, and that they form a complex involved in the exocytosis of bacteria. This capacity of TLR4 to mediate the expulsion of intracellular bacteria from infected cells represents a previously unrecognized function for this innate immune receptor.