IL-4 up-regulates cyclooxygenase-1 expression in macrophages

Abstract Macrophages use various cell-surface receptors to sense their environment and undergo polarized responses. The cytokines IL-4 and IL-13, released from T-helper type 2 (Th2) cells, drive macrophage polarization toward an alternatively activated phenotype (M2). This phenotype is associated with the expression of potent pro-resolving mediators, such as the prostaglandin (PG) D2-derived cyclopentenone metabolite, 15d-PGJ2, produced by the cyclooxygenase (Ptgs; Cox) pathway. Interestingly, IL-4 treatment of bone marrow-derived macrophages significantly downregulates Cox-2 protein expression, while Cox-1 levels are significantly increased. This phenomenon not only challenges the dogma that Cox-1 is only developmentally regulated, but also demonstrates a novel mechanism in which IL-4-dependent regulation of Cox-1 involves the activation of the mechanistic target of rapamycin (mTOR) complex. Using specific chemical inhibitors, we demonstrate here that IL-4-dependent Cox-1 upregulation occurs at the post-transcriptional level via the Fes-Akt-mTOR axis. Activation of AMP-activated protein kinase (Ampk) by metformin, inhibition of mTOR by torin, or CRISPR/Cas9-mediated genetic knockout of tuberous sclerosis-2 (Tsc2) blocked the IL-4-dependent expression of Cox-1 and ability of macrophages to polarize to M2. However, use of 15d-PGJ2 partially rescued the effects of Ampk activation, suggesting the importance of Cox-1 in macrophage polarization as also observed in a model of gastrointestinal helminth clearance. In summary, these findings suggest a new paradigm where IL-4-dependent upregulation of Cox-1 expression may play a key role in tissue homeostasis and wound healing during Th2-mediated immune responses, such as parasitic infections.