The Human G Protein-Coupled ATP Receptor P2Y₁₁ Is Associated With IL-10 Driven Macrophage Differentiation

The G protein-coupled P2Y₁₁ receptor is known to sense extracellular ATP during inflammatory and immune responses. The dinucleotide NAD⁺ has also been proposed to be a P2Y₁₁ receptor ligand but its role is less clear. Here, we have examined for the first time human P2Y₁₁ receptor protein levels and show that the receptor was upregulated during polarization of M2 macrophages. IL-10 reinforced P2Y₁₁ receptor expression during differentiation of M2c macrophages expressing CD163, CD16, and CD274 (PD-L1). Nutlin-3a mediated p53 stabilization further increased P2Y₁₁ receptor, CD16, and PD-L1 expression. AMP-activated kinase (AMPK), which mediates anti-inflammatory effects of IL-10, and nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme of the NAD⁺ salvage pathway, which is under the control of AMPK, were also required for P2Y₁₁ receptor expression. The P2Y₁₁ receptor agonist ATPγS and NAD⁺ could independently stimulate the production of IL-8 in M2 macrophages, however, only the ATPγS-induced response was mediated by P2Y₁₁ receptor. Both in a recombinant system and in macrophages, P2Y₁₁ receptor-driven IL-8 production predominantly depended on IkB kinase (IKK), and extracellular signal-regulated kinase (ERK). In conclusion, our data indicate that an AMPK-NAMPT-NAD⁺ signaling axis promotes P2Y₁₁ receptor expression during M2 polarization of human macrophages in response to IL-10. PD-L1 expressing M2c macrophages that secrete the cancer-promoting chemokine IL-8 in response to P2Y₁₁ receptor stimulation may represent an important target in checkpoint blockade immunotherapy.