Prior infection of Galleria mellonella with sublethal dose of Bt elicits immune priming responses but incurs metabolic changes.

Invertebrate immune priming has attracted wide attention of biologists in recent years because it challenges core notions about the disparate nature of acquired and innate immunity. However, the metabolic switch and energetic cost during eliciting immune priming are poorly investigated issues, which could widen and deepen our understanding of the physiological mechanism of immune priming. In this study, using sublethal dose of Bacillus thuringiensis (Bt) as an elicitor, we detected typical immune priming responses in Galleria mellonella. We found that the intensity of immune priming is positively correlated with the levels of antimicrobial peptides and phagocytosis ability of hemocytes. Subsequently, we employed LC-MS/MS-based untargeted metabolomics techniques to analyze the metabolic changes in the fat body of G. mellonella larvae during immune priming. The results showed that there were 74 and 56 significantly altered metabolites in positive and negative ion mode, respectively, after Bt priming. Most of the differential metabolites were enriched in the following metabolic pathways: amino acid biosynthesis, carbon metabolism, aminoacyl-tRNA biosynthesis and ABC transporters. The energetic cost of immune priming was depicted mainly in the slow growth of body mass and decreased levels of sucrose, lactose, D-ribulose 1,5-bisphosphate, Glycerate-3P and isocitric acid, which are enriched in carbon metabolism and involved in energy production. Meanwhile, correlation and interaction network analysis showed negative correlations between carbohydrates and metabolites involved in amino acid biosynthesis, suggesting that amino acids acted as the main energy source and helped the organisms synthesize immune effectors to participate in the immune priming response. Our results pave the way for uncovering the physiological mechanism of insect immune priming and discovering novel targets for Bt insecticide.