Age-Dependent Enterocyte Invasion and Microcolony Formation by Salmonella

Abstract

The coordinated action of a variety of virulence factors allows Salmonella enterica to invade epithelial cells and penetrate the mucosal barrier. The influence of the age-dependent maturation of the mucosal barrier for microbial pathogenesis has not been investigated. Here, we analyzed Salmonella infection of neonate mice after oral administration. In contrast to the situation in adult animals, we observed spontaneous colonization, massive invasion of enteroabsorptive cells, intraepithelial proliferation and the formation of large intraepithelial microcolonies. Mucosal translocation was dependent on enterocyte invasion in neonates in the absence of microfold (M) cells. It further resulted in potent innate immune stimulation in the absence of pronounced neutrophil-dominated pathology. Our results identify factors of age-dependent host susceptibility and provide important insight in the early steps of Salmonella infection in vivo. We also present a new small animal model amenable to genetic manipulation of the host for the analysis of the Salmonella enterocyte interaction in vivo.

Author Summary

Non-typhoidal Salmonella are among of the most prevalent causative agents of infectious diarrheal disease worldwide but also very significantly contribute to infant sepsis and meningitis particularly in developing countries. The underlying mechanisms of the elevated susceptibility of the infant host to systemic Salmonella infection have not been investigated. Here we analyzed age-dependent differences in the colonization, mucosal translocation and systemic spread in a murine oral infection model. We observed efficient entry of Salmonella in intestinal epithelial cells of newborn mice. Enterocyte invasion was followed by massive bacterial proliferation and the formation of large intraepithelial bacterial colonies. Intraepithelial, but not non-invasive, extracellular Salmonella induced a potent immune stimulation. Also, enterocyte invasion was required for translocation through the mucosal barrier and spread of Salmonella to systemic organs. This requirement was due to the absence of M cells, specialized epithelial cells that forward luminal antigen to the underlying immune cells, in the neonate host. Our results identify age-dependent factors of host susceptibility and illustrate the initial phase of Salmonella infection. They further present a new small animal model amenable to genetic manipulation to investigate the interaction of this pathogen with epithelial cells and characterize the early steps in Salmonella pathogenesis.