Mitochondrial dysfunction and glycolytic shift define the metabolic profile of hydatid fluid-stimulated dendritic cells.

Dendritic cells (DCs) require substantial metabolic reprogramming to mount an immune response against parasites. In the context of Echinococcus granulosus infection, parasite antigens can modulate host immune responses; however, their specific impact on DC metabolism remains poorly defined. We measured mitochondrial membrane potential (ΔΨM), intracellular reactive oxygen species (ROS), and nitric oxide (NO) production in FMS-like tyrosine kinase 3 ligand (FLT3-L)-derived DC (FL-DCs) stimulated with hydatid fluid (HF) or purified laminar layer (pLL) from E. granulosus. Gene expression profiling of key metabolic enzymes involved in glycolysis and oxidative phosphorylation (OXPHOS) was performed using RT-qPCR. Functional metabolic flux was analyzed using the Seahorse glycolysis stress test, while key metabolites from both pathways were quantified by HPLC. Our results showed that HF-stimulated FL-DCs exhibited marked mitochondrial dysfunction, evidenced by reduced ΔΨM and diminished mitochondrial network complexity, in contrast to the preserved mitochondrial morphology in pLL-stimulated cells. Gene expression analysis revealed that both stimuli enhanced glycolytic enzyme transcripts; however, only pLL induced OXPHOS-related genes, suggesting divergent bioenergetic adaptations. HF-stimulated FL-DCs produced elevated levels of ROS and NO, indicative of oxidative stress and a glycolysis-favored metabolic state, whereas pLL maintained mitochondrial respiration without excessive ROS production. Metabolic flux assays corroborated these findings; HF-stimulated cells displayed an increased extracellular acidification rate and a decreased oxygen consumption rate, indicating a glycolytic shift. In contrast, pLL-stimulated FL-DCs preserved oxidative metabolism and aerobic glycolysis. Our findings demonstrate that HF and pLL from E. granulosus differentially modulate metabolic programs in FL-DCs and highlight potential targets for modulating DC function in echinococcosis pathogenesis.