TLR8 activation shifts neutrophils from phagocytosis to NETosis through furin-dependent shedding of FcgRIIA

Background Immune complex (IC)-mediated formation of neutrophil extracellular traps (NETs) has emerged as a mechanism to increase the autoantigenic burden as well as promote the prominent type I interferon (IFN) signature seen in patients with the autoimmune disease systemic lupus erythematosus (SLE). The aim of the current study was to explore properties of ICs regulating their ability to induce phagocytosis and NETosis. Results Whereas immobilized IgG, mimicking tissue-deposited ICs, induced NETs in a TLR-independent manner, both FcgR- and TLR-engagement were required for induction of NETosis by soluble RNA-ICs, as demonstrated by FcgR blocking antibodies as well as RNase-treatment. Despite that degradation of RNA inhibited NETosis, removal of the TLR ligand by RNase markedly increased the phagocytosis of RNA-ICs by neutrophils suggesting that TLR activation suppressed phagocytosis. Consistent with this hypothesis, addition of a TLR8 agonist (R848) inhibited phagocytosis of ICs, but not beads, in neutrophils. Mechanistically, we observed that TLR8 induced ROS-dependent activation of the serine protease furin, participating in shedding of cell surface FcgRIIA reducing the phagocytic capacity but promoting progression into NETosis. Conclusions i) In the fluid phase, both FcgRIIA and TLR8 ligation are necessary for IC-mediated NETosis; ii) TLR8 activation induces furin-mediated shedding of FcgRIIA, thereby inhibiting further phagocytosis of ICs but enabling NETosis. Therapeutic approaches aimed at degrading the TLR8 ligand, such as RNases, would be predicted to increase uptake of circulating ICs, while disarming their inflammatory potential and ability to induce NETs.