Mitochondrial translation inhibition in CD4+T cells selectively sensitizes the Th2 subset to increased cell death

Abstract

Abstract Upon stimulation with antigen, resting T cells undergo extensive proliferation followed by differentiation into effector cells and long-lived memory cells. Upregulation of mitochondrial oxidative phosphorylation (OXPHOS) is essential for the activation and proliferation of resting T cells. Mitochondrial DNA encodes 13 proteins, which are integral parts of the complexes that form the mitochondrial respiratory chain. Since mitochondrial ribosomes are evolutionarily similar to bacterial ribosomes, antibiotics targeting bacterial ribosomes inhibit mitochondrial protein synthesis as well. But how these antibiotics affect T cell proliferation and differentiation is not well understood. To address this, we used chloramphenicol to inhibit mitochondrial translation during the activation of naive murine CD4+T cells in Th1 or Th2 polarizing conditions. Cell proliferation and effector functions were reduced in both Th1 and Th2 cells when activated in presence of chloramphenicol. Moreover, chloramphenicol significantly altered metabolic reprogramming in Th1 and Th2 cells indicated by a lower rate of OXPHOS and upregulation of glycolysis to maximum levels. Interestingly, inhibition of mitochondrial translation resulted in increased apoptosis following activation specifically in the Th2 effector subset. Th2 cells displayed a significant increase in caspase 3/7 activity upon mitochondrial translation inhibition. RNA sequencing analysis identified the upregulation of Bim and Puma in Th2 cells, two pro-apoptotic proteins that are known to be upregulated in response to metabolic stress. These results show that Th2 cells specifically undergo apoptosis in response to antibiotics that target mitochondrial translation. Support from ICMR, NII-DBT