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

ABSTRACT 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 BBP2.1DWV and BBP2.1IAPV. 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.IMPORTANCEEach year, approximately 40% of managed honey bee hives in the United States are lost due to a variety of environmental stressors. Although increases in virus infection are among the most important factors resulting in colony loss, there are currently no effective tools for the management of virus infection in honey bees. In this study, we identified a peptide that binds to the gut of the honey bee and competes with two of the most important honey bee viruses, Israeli acute paralysis virus of bees (IAPV) and Deformed wing virus (DWV), for binding to gut proteins. In vivo competition between this peptide and DWV demonstrates the potential utility of gut-binding peptides for the protection of honey bees from virus infection for reduced virus-associated honey bee mortality.