Basal interferon signaling and therapeutic use of interferons in controlling rotavirus infection in human intestinal cells and organoids

Mohamad S. Hakim,Qiuwei Pan,Aqsa Ikram,Shihao Ding,Sunrui Chen,Wenshi Wang,Maikel P. Peppelenbosch,Xiao-xia Ma,Yuebang Yin

doi:10.1038/s41598-018-26784-9

Abstract

Rotavirus (RV) primarily infects enterocytes and results in severe diarrhea, particularly in children. It is known that the host immune responses determine the outcome of viral infections. Following infections, interferons (IFNs) are produced as the first and the main anti-viral cytokines to combat the virus. Here we showed that RV predominantly induced type III IFNs (IFN-λ1), and to a less extent, type I IFNs (IFN-α and IFN-β) in human intestinal cells. However, it did not produce detectable IFN proteins and thus, was not sufficient to inhibit RV replication. In contrast, we revealed the essential roles of the basal IFN signaling in limiting RV replication by silencing STAT1, STAT2 and IRF9 genes. In addition, exogenous IFN treatment demonstrated that RV replication was able to be inhibited by all types of IFNs, both in human intestinal Caco2 cell line and in primary intestinal organoids. In these models, IFNs significantly upregulated a panel of well-known anti-viral IFN-stimulated genes (ISGs). Importantly, inhibition of the JAK-STAT cascade abrogated ISG induction and the anti-RV effects of IFNs. Thus, our study shall contribute to better understanding of the complex RV-host interactions and provide rationale for therapeutic development of IFN-based treatment against RV infection.

Highlights

Rotavirus (RV) is a member of the Reoviridae family that primarily infects mature enterocytes of the small intestinal villi
An effective replication was shown by an increase in intracellular RNA level as well as secreted rotavirus in culture medium (Supplementary Fig. S1)
To rule out the possibility that endogenous IFN produced following RV infection is sufficient to restrict RV replication, we investigated whether the inhibition of JAK proteins, the downstream elements of IFN receptor, influences RV replication

Summary

Introduction

Rotavirus (RV) is a member of the Reoviridae family that primarily infects mature enterocytes of the small intestinal villi. Some members are widely used in the clinic for treating viral infections or malignancy; whereas others are at stages of clinical development. Even though they bind to distinct receptors, they signal through a common, classical Janus kinase signal transducer and activator of transcription (JAK-STAT) pathway[8,9]. Viral nonstructural protein NSP1-mediated IFN inhibition has been shown to be associated with different levels of RV replication in primary mouse cells[15]. Mice deficient in type I or II IFN receptor signaling controlled RV infection as wild-type mice[24,25] These results suggest a minor role of type I and II IFNs in controlling RV infection in mice. Administration of IFN-λ conferred better protection against RV infection than IFN-α/β25

Methods

Results

Conclusion