TARGETING CD4+ T CELL METABOLISM PROVIDES AN EFFECTIVE AND AGE-SPECIFIC IMMUNOSUPPRESSION

Abstract

Targeting T-cell metabolism may provide an effective immunosuppression. We delineated the correlation of aging and T cell metabolism. Moreover, we defined the consequences of metabolic changes in T-cell compartments on alloimmunity and delineated specific and novel immunosuppressive targets on CD-4+ T-cells. We collected naïve CD4+ T cells (splenocytes) of young and old (3 and 18 months) C67BL/6 mice. Following activation with anti-CD3 and anti-CD28 (24 hrs), we evaluated oxidative phosphorylation (OXPHOS) and aerobic glycolysis by oxygen consumption (OCR) and extracellular acidification (ECAR) using a Seahorse XFe96 extracellular flux analyzer. Graft survival, T-cell specificity and a detailed immune profiling were recorded serially. Upon activation in-vitro, a compromised utilization of OXPHOS and glycolysis pathways was observed in old CD4+ T cells (p<0.0001), indicating failure of old CD4+ T cells to exhibit adequate metabolic reprogramming. Further analysis of metabolic pathways revealed a critical role of glutaminolysis in T cell activation. 6-diazo-5-oxo-l-norleucine (DON), an analog of glutamine inhibiting critical enzymes of glutaminolysis resulted into an age-specific reduced activation and proliferative capacities of CD4+T cells. Notably, lower doses of DON inhibited IL-2 production and CFSE-determined proliferation only in old but not young CD4+ T cells (p<0.001). Utilizing a pre-clinical fully mismatched transplantation model, we observed inversed CD4:CD8 ratio in old recipients following DON (1.6mg/kg every other day). Graft survival was prolonged in young recipients, from 7 to 18 days (p< 0.01). Notably, was a drastic and age-specific effect of DON on graft skin graft survival in old recipients to 42 days (n=7, p<0.001). Targeting metabolic pathways was specific to CD4+ T-cells. Adoptively transferring young CD4+ T cells reversed the prolonged survival observed in old recipients. Depletion of CD8+ T cells did not alter transplant outcomes of DON in either young or old recipients. Taken together, we introduce a detailed and age-specific analysis of metabolic pathways in T cells and introduce a novel concept for an optimized immunosuppression in old recipients.