Inhibiting infectious bronchitis virus PLpro using ubiquitin variants

Abstract

Infectious bronchitis virus (IBV) is a coronavirus first isolated in the 1930s infecting chickens. IBV causes great economic losses to the global poultry industry, as it affects egg production and causes mortality by leaving the host susceptible to secondary bacterial infections. Even though vaccines are available, they are poorly cross-protective against new variants of the virus, which are always on the cusp of emerging. Effective antiviral therapies, or possibly the production of transgenic animals immune to IBV infection, are therefore sorely needed. As the papain-like protease (PLpro) of IBV has deubiquitinating activity besides its crucial ability to cleave the viral polyprotein, we have applied a novel strategy of selecting ubiquitin variants (UbVs) from a phage-displayed library that have high affinity to this viral protease. These UbVs were found to inhibit the deubiquitinating activity of PLpro and consequently obstruct the virus's ability to evade the innate immune response in the host cell. By obstructing the proteolytic activity of PLpro, these UbVs were seemingly able to inhibit viral infection as assessed using immunofluorescence microscopy. Whilst virus infection was detected in around 5% of UbV-expressing cells, the virus was present in around 30-40% of GFP (control)-expressing cells. This suggests that the expression of UbVs indeed seems to inhibit IBV infection, making UbVs a potentially potent and innovative antiviral strategy in the quest for control of IBV infections.