Enteroviruses infect human enteroids and induce antiviral signaling in a cell lineage-specific manner

Abstract

Enteroviruses are among the most common viral infectious agents of humans and are primarily transmitted by the fecal-oral route. However, the events associated with enterovirus infections of the human gastrointestinal tract remain largely unknown. Here, we used stem cell-derived enteroids from human small intestines to study enterovirus infections of the intestinal epithelium. We found that enteroids were susceptible to infection by diverse enteroviruses, including echovirus 11 (E11), coxsackievirus B (CVB), and enterovirus 71 (EV71), and that contrary to an immortalized intestinal cell line, enteroids induced antiviral and inflammatory signaling pathways in response to infection in a virus-specific manner. Furthermore, using the Notch inhibitor dibenzazepine (DBZ) to drive cellular differentiation into secretory cell lineages, we show that although goblet cells resist E11 infection, enteroendocrine cells are permissive, suggesting that enteroviruses infect specific cell populations in the human intestine. Taken together, our studies provide insights into enterovirus infections of the human intestine, which could lead to the identification of novel therapeutic targets and/or strategies to prevent or treat infections by these highly clinically relevant viruses.E nteroviruses are significant sources of human infections worldwide and are primarily transmitted by the fecal-oral route. Nonpoliovirus enteroviruses include coxsackievirus, echovirus, enterovirus 71 (EV71), and enterovirus D68 (EV-D68) and are small (∼30 nm) single-stranded RNA viruses belonging to the Picornaviridae family. In many cases, enterovirus infections remain asymptomatic, whereas in others, infection is associated with mild flu-like symptoms or much more severe outcomes such as type I diabetes, encephalomyelitis, encephalitis, myocarditis, dilated cardiomyopathy, pleurodynia, acute flaccid paralysis, or even death.The human gastrointestinal (GI) tract is a complex organ, with an epithelial surface that must provide a protective and immunological barrier in a complex and diverse microbial environment. The epithelium of the small intestine contains at least seven distinct cell subtypes that are responsible for the physiological functions of the intestine, which include nutrient absorption and defense against pathogens. The lack of models that recapitulate the complexity of the GI tract has hindered studies into many aspects of enterovirus infection in this specialized environment. Although murine models have been developed for the study of enterovirusinduced disease (1-4), many of these models require intraperitoneal (i.p.) infection, thereby bypassing the GI tract, or require ablation of the host innate immune system (5, 6). Coupled with species differences between humans and mice, there remains a need to develop human-based platforms to model enterovirus infections of the GI tract.The full repertoire of mature cells in the small intestine in vivo includes those of absorptive (enterocytes) and secretory (Paneth, goblet, and enteroendocrine) lin...

Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses

“…Rotavirus genotype P [7] has been classified as a sialic acid-dependent genotype 43 , which may explain why rotavirus G1P [7] strain could also infect colon epithelial cells in vitro. Both ileum and colon epithelial cells were also susceptible to four fecal suspensions containing different rotavirus genotypes (G5P [13], G5P [7], G9P [23] and G4P [6]). G5P [7] and G9P [23] rotaviruses, which are the predominant G/P genotype combination of circulating rotaviruses in Belgium, were more infectious than the other two strains, which was also observed on MA104 cells 8 .…”

“…Both ileum and colon epithelial cells were also susceptible to four fecal suspensions containing different rotavirus genotypes (G5P [13], G5P [7], G9P [23] and G4P [6]). G5P [7] and G9P [23] rotaviruses, which are the predominant G/P genotype combination of circulating rotaviruses in Belgium, were more infectious than the other two strains, which was also observed on MA104 cells 8 . The higher infectivity in enterocytes in vitro may explain the wide prevalence of these genotypes in the field.…”

“…The results showed that these four genotypes of rotavirus could infect both ileum and colon epithelial cells. Rotavirus 14R163 (G9P [23]) strain demonstrated the highest infectivity to primary enterocytes, followed by the 14R160 (G5P [7]) strain. For rotavirus 14R165 (G4P [6]) strain, an increase of more than 10 fold RNA copies/ml was detected at 24 h post inoculation of ileum and colon epithelial cells.…”

“…Intestinal enteroids have been developed by culturing intestinal crypts onto Matrigel which is enriched with laminin α1 and α2 20 . Enteroids enhanced the viability of the cells and were already used for the study of rotaviruses 21 , noroviruses 22 and enteroviruses 23 . The successful cultivation of enteroids is dependent on many growth factors, critically including Wnt 3a, R-spondin, and Noggin, which is an expensive method.…”

Modeling Host-Pathogen Interactions in the Context of the Microenvironment: Three-Dimensional Cell Culture Comes of Age

“…Successful viral replication was observed and only enterocytes were infected with HuNoVs, regardless of the strain or the intestinal region from which the model was derived. Additional viral models, including those using enteroviruses (e.g., CVB, echovirus 11, and enterovirus 71) have identified the cell-type-specific nature of these infections and the virus-specific nature of innate immune signaling in response to infection (327).…”