Transcriptome analysis of pig intestinal cell monolayers infected with Cryptosporidium parvum asexual stages

Abstract

BackgroundHuman cryptosporidiosis is caused primarily by two species of apicomplexan protozoa, Cryptosporidium parvum and C. hominis. In cultured cell monolayers, the parasite undergoes two generations of asexual multiplication (merogony). However, the proportion of parasites completing the life-cycle is low and insufficient to sustain continuous propagation. Due to the intracellular location of meronts and later life-cycle stages, oocyst and sporozoites are the only forms of the parasite that can readily be isolated.ResultsResearch on the replicating forms of Cryptosporidium parasites and their interaction with the host cell remains challenging. Based on an RNA-Seq analysis of monolayers of pig epithelial cells infected with C. parvum, here we report on the impact of merogony on the host’s gene regulation. Analysis of the transcriptome of infected and uninfected monolayers demonstrates a significant impact of the infection on host cell gene expression. A total of 813 genes were differentially expressed. Functional terms significantly altered in response to infection include phosphoprotein, RNA binding and acetylation. Upregulation of cell cycle pathways indicates an increase in mitosis. Notably absent from differentially enriched functional categories are stress- and apoptosis-related functions. The comparison of the combined host-parasite transcriptome reveals that C. parvum gene expression is less diverse than the host cell transcriptome and is highly enriched for genes encoding ribosomal functions, such as ribosomal proteins.ConclusionsThese results indicate that C. parvum infection significantly changes host biological functions and provide new insight into gene functions driving early C. parvum intracellular development.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-2754-3) contains supplementary material, which is available to authorized users.

Transcriptomic Study of Porcine Small Intestine Epithelial Cells Reveals Important Genes and Pathways Associated With Susceptibility to Escherichia coli F4ac Diarrhea

“…Our results found a total of 543 DEGs (P < 0.05 and log 2 |FC| > 1) in comparison of ETEC-F4ac non-adhesive vs. adhesive small intestine epithelial cells ( Figure 1E) and a total of 822 DEGs with P < 0.05 ( Supplementary Table S3). A previous study profiled the transcriptome of IPEC-J2 cell monolayers found 810 DEGs (FDR < 0.05) (Mirhashemi et al, 2018).…”

“…Nowadays, high-throughput RNA sequencing (RNA-seq) technique has become a powerful tool and a standard method for the measurement and comparison of genes expression levels (Pertea et al, 2016). One of the most biologically relevant applications of RNA-Seq is the comparison of mRNA transcriptomes between infected and healthy individuals (Mirhashemi et al, 2018;Barroso and McCarthy, 2019). The Large White pig is a domestic pig breed originating in Yorkshire in the UK and currently is mostly reared commercial pig worldwide.…”

Recent Breakthroughs and Ongoing Limitations in Cryptosporidium Research

“…This study identified genes involved in ribosome biogenesis and translation as being upregulated in C. parvum during infection in vitro compared with oocysts. In addition, cell division pathways were upregulated in infected versus uninfected IPEC-J2 cells 40 . C. parvum transcripts accounted for only 2.2% of the total reads in this study, but whether that proportion was sufficient to measure low-level transcripts was not addressed.…”

Primary Cell-Derived Intestinal Models: Recapitulating Physiology

“…In vivo, the intestinal epithelium participates in extensive crosstalk with the neighboring environment such as luminal microorganisms, adjacent stroma, and immune cells. Epithelial monolayers provide a simplified tool to study these complex interactions in a controlled manner, and coculture with fibroblasts, bacteria, viruses, protozoa, neurons, and immune cells have all been demonstrated in recent years [66][67][68][69][70][71][72][73]. Placement of fibroblast feeder layers yields a layered tissue suitable for long-term epithelial culture or drug absorption and toxicity assays (Figure 3) [66,70,74].…”

“…The intestines play a central role in the maturation of the immune system, and macrophages cocultured with monolayers have demonstrated a coordinated epithelial-immune cell response to pathogens [72]. Perhaps one of the highest-value attributes of the monolayer culture systems is the ability to pair them with infectious agents such as noroviruses [67], adenoviruses [68], enterohemorrhagic E. coli [69], or Cryptosporidium parasites [71] to reveal the intricate details of host-pathogen interactions.…”