To gain insight into the mechanism of Lactobacillus acidophilus (L. acidophilus) S-layer protein antiviral activity, we examined how S-layer protein impacts porcine epidemic diarrhea virus (PEDV) infection and PEDV-induced apoptosis of Vero cells. Pretreatment (exclusion assay), coincubation (competition assay), and post-treatment (displacement assay) of PEDV-infected Vero cells with the S-layer protein was examined. Interestingly, significant inhibition of PEDV by S-layer protein was only observed in the exclusion assay. In Vero cells infected with PEDV, we found that apoptosis was mediated by activation of caspase-8 and caspase-3 in the late stage of infection. When PEDV-infected Vero cells were pretreated with S-layer protein, rates of Vero cell apoptosis were markedly decreased and cell damage was significantly reduced, as evaluated by flow cytometry and microscopy. Detailed analyses showed that the S-layer protein inhibited caspase-8 and caspase-3 activity. Taken together, our results suggest that L. acidophilus S-layer protein plays an inhibitory role during PEDV infection of Vero cells, and that the antagonistic activity of the protein is not via competition with PEDV for binding sites. In addition, the findings suggest that L. acidophilus S-layer protein protects against PEDV-induced apoptosis through reduced caspase-8 and caspase-3 activation in the later stages of infection. This mechanism may represent a novel approach for antagonizing PEDV and other viruses.
Lactobacillus acidophilus S-layer protein-mediated inhibition of PEDV-induced apoptosis of Vero cells