Abstract
Several alphaherpesviruses breach the basement membrane during mucosal invasion. In the present study, the role of proteases in this process was examined. The serine protease-specific inhibitor AEBSF inhibited penetration of the basement membrane by the porcine alphaherpesvirus pseudorabies virus (PRV) by 88.1% without affecting lateral spread. Inhibitors of aspartic-, cysteine-, and metalloproteases did not inhibit viral penetration of the basement membrane. Further analysis using the Soybean Type I-S trypsin inhibitor for the serine protease subcategory of trypsin-like serine proteases resulted in a 96.9% reduction in plaque depth underneath the basement membrane. These data reveal a role of a trypsin-like serine protease in PRV penetration of the basement membrane.
Highlights
Several alphaherpesviruses breach the basement membrane during mucosal invasion
Using porcine nasal respiratory mucosal explants, we have previously confirmed that the porcine alphaherpesvirus pseudorabies virus (PRV) efficiently breaches the basement membrane (BM) [3]
In the current study, we investigated whether proteases are involved in alphaherpesvirus invasion through the BM
Summary
Several alphaherpesviruses breach the basement membrane during mucosal invasion. In the present study, the role of proteases in this process was examined. Incubation of PRV-inoculated explants with the serine-, cysteine- and metalloprotease inhibitor cocktail resulted in a 94.9% reduction in distance covered underneath the BM. PRV-inoculated explants were treated with 100 or 250 μM AEBSF, 1 or 10 μM E-64 or 10 μM phosphoramidon and the effect on the plaque depth underneath the BM and the plaque latitude above the BM was evaluated.
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