340 Tlr8 activation in neutrophils impairs immune complex phagocytosis through furin-dependent shedding of fcgriia

Background and aims Neutrophils play a crucial role in host defense through mechanisms including phagocytosis and formation of neutrophil extracellular traps (NETs), a recently identified neutrophil cell death process in which DNA is extruded together with cytoplasmic and granular content to trap and eliminate extracellular pathogens. Although beneficial from a host-pathogen perspective, exaggerated neutrophil activation has been linked to autoimmunity, in particular the rheumatic disease systemic lupus erythematosus (SLE) where nucleic acid-containing immune complexes (IC) drive inflammation. Toll-like receptor (TLR) agonists, such as nucleic acids, are important components of pathogens, enabling enhanced phagocytosis by macrophages and dendritic cells, but the role of TLR signalling in processing of SLE ICs and downstream inflammatory neutrophil effector functions is not known. Methods Standard Methods. Results We observed that both FcgR- and TLR8-engagement were required for induction of NETosis, whereas TLR8 activation, through the RNA component of the ICs, suppressed further IC-mediated phagocytosis. Mechanistically, TLR8 ligation induced PI3K-dependent ROS generation through NADPH oxidase, and subsequent furin-dependent proteolytic cleavage of the N-terminal part of FcgRIIA shifting neutrophils away from phagocytosis of ICs toward NETosis. TLR8 activated neutrophils promoted cleavage of FcgRIIA also on plasmacytoid dendritic cells and monocytes resulting in impaired overall clearance of ICs and increased complement C5a generation. Importantly , ex vivo derived activated neutrophils from SLE patients demonstrated a similar cleavage of FcgRIIA that was correlated with markers of disease activity as well as complement activation. Conclusions Therapeutic approaches aimed at blocking TLR8 activation would be predicted to increase phagocytosis of circulating ICs while disarming their inflammatory potential.