Neutrophil priming that turns natural FFA2R agonists into potent activators of the superoxide generating NADPH‐oxidase

Acetate, an agonist for the free fatty acid receptor 2 (FFA2R/GPR43), triggers an increase in the cytosolic concentration of free Ca2+in neutrophils without any assembly of the superoxide generating NADPH‐oxidase. We show that the phenylacetamide compound 58 (Cmp 58; (S)‐2‐(4‐chlorophenyl)‐3,3‐dimethyl‐N‐(5‐phenylthiazol‐2‐yl)butanamide), lacking a direct activating effect on neutrophils, acts as a positive FFA2R modulator that turns acetate into a potent activating agonist that triggers an assembly of the NADPH‐oxidase. The NADPH‐oxidase activity could be further increased in neutrophils treated with the pro‐inflammatory cytokine TNF‐α. Many neutrophil chemoattractant receptors are stored in secretory organelles but no FFA2R mobilization was induced in neutrophils treated with TNF‐α. The receptor selectivity was demonstrated through the inhibition of the neutrophil response induced by the combined action of acetate and Cmp 58 by the FFA2R antagonist CATPB. Receptor modulators that positively co‐operate with natural FFA2R agonists and prime neutrophils in their response to such agonists, may serve as good tools for further unraveling the physiological functions of FFA2R and its involvement in various diseases. In this study, we show that neutrophils primed with a presumed allosteric FFA2R modulator produce increased amounts of reactive oxygen species when activated by receptor specific agonists. Allosteric modulation of FFA2R, a novel receptor selective mechanism, primes neutrophils to produce increased amounts of reactive oxygen species.