Recent declines in honey bee (Apis mellifera L., 1758) populations worldwide have spurred significant research into the impact of pathogens on colony health. The role of the Israeli acute paralysis virus (IAPV) on hive mortality has become of particular concern since being correlated with colony losses. However, the molecular interactions between IAPV and its host remain largely unknown. To investigate changes in host protein expression during IAPV infection, mass-spectrometry-based quantitative proteomics was used to compare IAPV-infected and healthy pupae. Proteins whose expression levels changed significantly during infection were identified and functional analysis was performed to determine host systems and pathways perturbed by IAPV infection. Among the A. mellifera proteins most affected by IAPV, those involving translation and the ubiquitin–proteasome pathway were most highly enriched and future investigation of these pathways will be useful in identifying host proteins required for infection. This analysis represents an important first step towards understanding the honey bee host response to IAPV infection through the systems-level analysis of protein expression.