Gut-binding peptides as potential tools to reduce virus binding to honey bee gut surface proteins

Colonies of the western honey bee, Apis mellifera, are severely impacted by a wide range of stressors, with Varroa mites and associated viruses being among the most serious threats to honey bee health. Viral load plays an important role in colony demise, with the iflavirus Deformed wing virus (DWV) and the dicistrovirus Israeli acute paralysis virus (IAPV) being of particular concern. By feeding adult honey bees on a phage display library to identify gut-binding peptides (R. Mishra, Y. Guo, P. Kumar, P. E. Cantón, C. S. Tavares, R. Banerjee, S. Kuwar, and B. C. Bonning, Curr Res Insect Sci, 1:100012, https://doi.org/10.1016/j.cris.2021.100012), we identified Bee midgut-Binding Peptide (BBP2.1), which shares 75% and 85% identity with regions on the DWV capsid protein and IAPV ORFx protein, respectively. These viral protein domains are likely to be instrumental in virus interaction with the honey bee gut. Pull-down assays with honey bee gut brush border membrane vesicles were used to confirm peptide-mCherry binding to the gut for BBP2.1 and the two similar virus-derived sequences, peptides BBP2.1 ^DWV and BBP2.1 ^IAPV. In vitro competition assays showed that all three peptides compete with both IAPV and DWV virions for binding to honey bee gut-derived brush border membrane vesicles, suggesting that the three peptides and the two viruses bind to the same proteins. Ingestion of BBP2.1 reduced the movement of DWV, but not IAPV from the honey bee gut into the body and did not rescue IAPV-associated mortality. These results are discussed in relation to the biological function of IAPV ORFx and the potential utility of virus-blocking peptides for suppression of virus infection to reduce virus load and virus-associated honey bee mortality.