A Multicellular In Vitro Model of the Human Intestine with Immunocompetent Features Highlights Host‐Pathogen Interactions During Early Salmonella Typhimurium Infection

Abstract Studying the molecular basis of intestinal infections caused by enteric pathogens at the tissue level is challenging, because most human intestinal infection models have limitations, and results obtained from animals may not reflect the human situation. Infections with Salmonella enterica serovar Typhimurium (STm) have different outcomes between organisms. 3D tissue modeling of primary human material provides alternatives to animal experimentation, but epithelial co‐culture with immune cells remains difficult. Macrophages, for instance, contribute to the immunocompetence of native tissue, yet their incorporation into human epithelial tissue models is challenging. A 3D immunocompetent tissue model of the human small intestine based on decellularized submucosa enriched with monocyte‐derived macrophages (MDM) is established. The multicellular model recapitulated in vivo‐like cellular diversity, especially the induction of GP2 positive microfold (M) cells. Infection studies with STm reveal that the pathogen physically interacts with these M‐like cells. MDMs show trans‐epithelial migration and phagocytosed STm within the model and the levels of inflammatory cytokines are induced upon STm infection. Infected epithelial cells are shed into the supernatant, potentially reflecting an intracellular reservoir of invasion‐primed STm. Together, the 3D model of the human intestinal epithelium bears potential as an alternative to animals to identify human‐specific processes underlying enteric bacterial infections.