A Conserved Metabolic–Oxidative Axis Underlies Immune Cell Cryo-vulnerability

SUMMARY Immunotherapy has emerged as a transformative approach for treating cancer and other diseases, yet its widespread deployment requires effective cryopreservation strategies to enable scalable global distribution. However, many immune cell types remain acutely vulnerable to freeze-thaw stress, and the underlying mechanisms of this cryo-vulnerability are not well defined. In this study, we combined metabolic flux analysis, ROS quantification, lipidomics, and preclinical xenograft models to investigate how metabolic state influences cryopreservation outcomes. We found that immune cell activation induces a metabolic shift characterized by elevated glucose utilization and excessive ROS production, leading to profound post-thaw loss of viability and function, as demonstrated by a ∼25% survival rate in natural killer (NK) cells. Targeted pretreatments—including inhibitors of glucose metabolism, antioxidants, and suppression of lipid peroxidation—restored post-thaw recovery to nearly 90% while preserving effector activity and antitumor efficacy. Similar protective effects were observed across αβ T cells, γδ T cells, and macrophages, defining a conserved metabolic–oxidative pathway of cryo-vulnerability and offering applicable strategies to enhance immune cell preservation. Competing Interest Statement The authors have declared no competing interest.