Non‐cytopathic bovine viral diarrhoea virus 2 induces autophagy to enhance its replication

Abstract

Background Bovine viral diarrhoea virus (BVDV) is an important viral pathogen that has an economic impact on the livestock industry worldwide. Autophagy is one of the earliest cell‐autonomous defence mechanisms against microbial invasion, and many types of viruses can induce autophagy by infecting host cells. Objectives The aim of this study was to identify the role of autophagy in the pathogenesis of non‐cytopathic (ncp) BVDV2 infection. Methods Madin–Darby bovine kidney (MDBK) cells were treated with ncp BVDV2, rapamycin, or 3‐methyladenine (MA) and ncp BVDV2 and then incubated at 37°C for 24 h. Cells were harvested, and the effects of autophagy were determined by transmission electron microscopy (TEM), confocal laser microscopy, western blotting and qRT‐PCR. Apoptotic analysis was also performed using western blotting and flow cytometry. Results In ncp BVDV2‐infected MDBK cells, more autophagosomes were observed by TEM, and the number of microtubule‐associated protein 1 light chain 3B (LC3B) with green fluorescent protein puncta was also increased. The ncp BVDV2‐infected cells showed significantly enhanced conversion of LC3‐I to LC3‐II, as well as upregulation of autophagy‐related proteins, including ATG5 and Beclin 1, and substantial degradation of p62/SQSTM1. These results are similar to those induced by rapamycin, an autophagy inducer. E2 protein expression, which is associated with viral replication, increased over time in ncp BVDV2‐infected cells. Inhibition of autophagy by 3‐MA in ncp BVDV2‐infected MDBK cells downregulated the expressions of LC3‐II, ATG5 and Beclin 1 and prevented the degradation of p62/SQSTM1. Moreover, the expressions of phosphorylated Akt and procaspase‐3 were significantly increased in ncp BVDV2‐infected cells. In addition, the mRNA level of protein kinase R (PKR) was significantly reduced in ncp BVDV2‐infected cells. Conclusions Our results demonstrate that ncp BVDV2 infection induced autophagy in MDBK cells via anti‐apoptosis and PKR suppression. Therefore, autophagy may play a role in establishing persistent infection caused by ncp BVDV.

Immune evasion strategies of bovine viral diarrhea virus

“…Both biotypes had the ability to induce autophagy in MDBK and primary bovine turbinate (Bt) cells at similar levels and used autophagy to replicate themselves ( Rajput et al., 2017 ). Similarly, ncp BVDV infection induced autophagy in MDBK cells by significantly increasing the conversion of LC3-I to LC3-II, upregulating the autophagy-related proteins ATG5 and Beclin 1 and degrading p62/SQSTM1, thereby promoting viral replication( Shin et al., 2023 ). The BVDV non-structural protein NS4B alone was able to induce autophagosomes in bovine kidney LB9.k cells, representing a novel and significant function of NS4B in BVDV replication ( Suda et al., 2019 ).…”