Apoptotic hemocytes induced by Microplitis bicoloratus parasitism have been reported, and M. bicoloratus bracovirus (MbBV) is known to be the apoptosis inducer. However, the mechanism how MbBV regulates apoptosis remains unclear. eIF4A, one of translation initiation factors, was found from a Spodoptera litura transcriptome, the expression of which in the parasitized hemocytes of S. litura was inhibited in RT‐qPCR analysis. The western blot also illustrated eIF4A at 6‐day post‐parasitization was inhibited in hemocytes. For testing interaction of MbBV–eIF4A–apoptosis, a cDNA clone encoding 1,266 bp of eIF4A was obtained from S. litura hemocytes and sequenced. Then, a 48 kDa V5‐fusion protein of the eIF4A was detected by using the anti‐V5 antibody at 72‐h post‐transfection in the High Five cells, which is located in the cell cytoplasm. In vitro, overexpression of eIF4A rescued the apoptotic High Five cells induced by MbBV. Conversely, in vivo, loss of eIF4A proteins by dsRNA feeding increased apoptosis of hemocytes. Furthermore, RNAi and parasitism significantly increased apoptosis of hemocytes in S. litura. These findings suggested that MbBV inhibited the expression of eIF4A, which was required for apoptosis mediated by MbBV. This study will contribute to biological pest control and enhance our understanding of molecular mechanisms underlying polydnavirus–parasitoid–host interaction.