Abstract
Senecavirus A (SVA) is a member of the genus Senecavirus of the family Picornaviridae. SVA-associated vesicular disease (SAVD) outbreaks have been extensively reported since 2014–2015. Characteristic symptoms include vesicular lesions on the snout and feet as well as lameness in adult pigs and even death in piglets. The capsid protein VP3, a structural protein of SVA, is involved in viral replication and genome packaging. Here, we developed and characterized a mouse monoclonal antibody (mAb) 3E9 against VP3. A motif 192GWFSLHKLTK201 was identified as the linear B-cell epitope recognized by mAb 3E9 by using a panel of GFP-tagged epitope polypeptides. Sequence alignments show that 192GWFSLHKLTK201 was highly conserved in all SVA strains. Subsequently, alanine (A)-scanning mutagenesis indicated that W193, F194, L196, and H197 were the critical residues recognized by mAb 3E9. Further investigation with indirect immunofluorescence assay indicated that the VP3 protein was present in the cytoplasm during SVA replication. In addition, the mAb 3E9 specifically immunoprecipitated the VP3 protein from SVA-infected cells. Taken together, our results indicate that mAb 3E9 could be a powerful tool to work on the function of the VP3 protein during virus infection.
Highlights
Senecavirus A (SVA) is a member of the genus Senecavirus of the family Picornaviridae, which is considered one of the etiological agents causing vesicular disease in pigs
Monoclonal antibodies of SVA VP3 were generated to investigate the distribution of VP3 in the SVA-infected cells, and a B cell epitope was finely mapped in this study
These results demonstrated that monoclonal antibody (mAb) recognized the SVA VP3 protein
Summary
Senecavirus A (SVA) is a member of the genus Senecavirus of the family Picornaviridae, which is considered one of the etiological agents causing vesicular disease in pigs. The VP3 protein of the Duck hepatitis A virus plays an important role in the host’s cell adsorption and apoptosis [14]. A recent study has shown that SVA VP3 protein is involved in cell autophagy [17]. These findings indicate that picornavirus VP3 proteins play critical roles in the viral infection of the host. Monoclonal antibodies (mAbs) of SVA VP3 were generated to investigate the distribution of VP3 in the SVA-infected cells, and a B cell epitope was finely mapped in this study. The results indicate that VP3-specific mAb will be useful in elucidating the function of VP3 during virus infection and developing new diagnostic reagents
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