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. 2018 Jul:120:32-36.
doi: 10.1016/j.micpath.2018.04.046. Epub 2018 Apr 24.

miR-146a-5p promotes replication of infectious bronchitis virus by targeting IRAK2 and TNFRSF18

Hui Liu  1 , Xin Yang  1 , Zhi-Kun Zhang  1 , Wen-Cheng Zou  1 , Hong-Ning Wang  2
Affiliations

miR-146a-5p promotes replication of infectious bronchitis virus by targeting IRAK2 and TNFRSF18

Hui Liu et al. Microb Pathog. 2018 Jul.

Abstract

Avian infectious bronchitis virus (IBV) is a coronavirus which infects chickens (Gallus gallus) of all ages and causes significant economic losses to the poultry industry worldwide. The present study aims to analyze the miRNAs related to pathogenicity of nephropathogenic IBVs. It was found that four miRNAs (miR-1454, miR-3538, miR-146a-5p and miR-215-5p) were related to the infection of virulent nephropathogenic IBV with transcript per million (TPM) > 500 and more than a 2-fold alteration. In vitro study results showed that the alterations of these four miRNAs were consistent with in vivo data. In vitro, we found that high levels of miR-146a-5p could enhance the replication of IBV at the early stage of infection, and its down regulated level could slow down the replication of IBV. Finally, high levels of exogenous miR-146a-5p in HD11 cells led to down regulation of IL-1 receptor associated kinase-2 (IRAK2) and Tumor necrosis factor receptor superfamily member 18 (TNFRSF18) genes. Luciferase reporter assays revealed that miR-146a-5p could bind to the 3'-UTRs of IRAK2 and TNFRSF18. This is the first study demonstrating that IBV induced miR-146a-5p is related to virus pathogenesis by down regulating IRAK2 and TNFRSF18, which may serve as a therapeutic strategy for the prevention of IBV infections.

Keywords: IBV; IRAK2; TNFRSF18; miR-146a-5p.

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Figures

Fig. 1
Fig. 1
miRNAs expression levels in HD11 and Vero cells infected with Beaudette. Both HD11 and Vero cells were infected with Beaudette, and then expression levels of miR-1454, miR-3538, miR-146a-5p, miR-215-5p were examined after 24 h of infection. **p < 0.01.
Fig. 2
Fig. 2
miR-146a-5p promote the replication of Beaudette at the early stage after infection.(a) Beaudette showed high titration in HD11 cells at 24 h after transfected with miR-146a-5p mimic, and miR-146a-5p inhibitor reduced the replication of Beaudette. (b & c) miR-146a-5p promote the replication of Beaudette at the early stage after infection, cytopathic effects exhibited at 33 h in miR-146a-5p transfected-infected cells compared to control cells (36 h).
Fig. 3
Fig. 3
miR-146a-5p target genes expression analysis. (a) The transcription levels of TNFRSF18 and IRAK2 were down regulated (p < 0.01) in miR-146a-5p transfected cells. (b) Predicted binding sites on the 3′-UTRs regions of TNFRSF18 and IRAK2 (c) Levels of TNFRSF18 and IRAK2 luciferase expression in HEK 293T cells transfected with miR-146a-5p (p < 0.01).
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