Cryptosporidium parvum infection attenuates the ex vivo propagation of murine intestinal enteroids

Abstract

Cryptosporidium, a ubiquitous coccidian protozoan parasite that infects the gastrointestinal epithelium and other mucosal surfaces, is an important opportunistic pathogen for immunocompromised individuals and a common cause of diarrhea in young children in the developing countries. One of the pathological hallmarks of intestinal cryptosporidiosis is villous atrophy, which results in a shorter height of intestinal villi. Here, we investigated the effects of Cryptosporidium infection on intestinal epithelial growth, using an ex vivo model of intestinal cryptosporidiosis employing enteroids from mice. We detected infection of enteroids isolated from immunocompetent adult and neonatal mice after ex vivo exposure to Cryptosporidium sporozoites. We observed a significant inhibition of enteroid propagation following infection. Intriguingly, we identified a decreased expression level of intestinal stem cell markers in enteroids following C. parvum infection. We further measured the expression levels of several Wnt antagonists or agonists in infected enteroids, as induction of the Wnt/β‐catenin activation is a key factor for intestinal stem cell function. We detected a markedly increased level of the Dickkopf‐related protein 1 and decreased level of the Wnt family member 5a in enteroids after infection. The low density lipoprotein receptor‐related protein 5, one of the Wnt co‐receptors, is downregulated in the infected enteroids. In addition, increased apoptotic cell death and cell senescence were observed in the infected enteroids. Our results demonstrate a significant inhibitory effect of Cryptosporidium infection on the ex vivo propagation of enteroids from mice, providing additional insights into the impact of Cryptosporidium infection on intestinal epithelial growth.

Development of in vitro enteroids derived from bovine small intestinal crypts

“…Enteroid cultures have been shown to have useful practical application in the study of the signalling pathways which modulate the differentiation and maintenance of distinct epithelial cell lineages in the murine intestine including Lgr5 + intestinal stem cells [ 6 , 61 ], tuft cells [ 20 ] and antigen-sampling M cells [ 3 , 19 , 21 , 22 ]. The 3D nature of the enteroids with a layer of epithelial cells surrounding a central, closed, lumen has also been shown to be an effective system in which to model mucosal permeability [ 62 ] and the interactions of mucosally-acquired pathogenic bacteria [ 24 ], viruses [ 25 ] and protozoa [ 26 ] with the gut epithelium. Our data suggest that these 3D bovine enteroid cultures represent a novel, physiologically-relevant and tractable in vitro system in which epithelial cell differentiation and function, and host–pathogen interactions in the bovine small intestine can be studied.…”

“…Enteroid cultures have been used in many laboratories to study cell differentiation and function in the intestinal epithelium [ 3 , 6 , 13 , 15 – 23 ]. The central closed lumen of enteroids also enables the uptake of particulate antigens [ 19 ] and host–pathogen interactions at the mucosal surface [ 24 – 26 ] to be studied.…”

Bovine cryptosporidiosis: impact, host-parasite interaction and control strategies

“…Zhang et al provided the first study of C. parvum and intestinal organoids, which were derived from mouse tissue. The authors illustrated that the presence of C. parvum attenuated the differentiation of murine intestinal organoids [ 156 ]. Therefore, similar in vitro systems would be advantageous to understand the nature of the host–pathogen interaction during C. parvum infection in cattle, and other relevant host species.…”

The Role of Ion Transporters in the Pathophysiology of Infectious Diarrhea

“… 67 , 68 Finally, C parvum causes a loss of absorptive villous enterocytes in the small intestine with an associated reduction in SGLT1 abundance and function, which has been assumed to underlie diarrheal pathogenesis along with host cell prostaglandin production and diminished barrier function. 69 , 70 , 71 …”