Modeled Microgravity Induces Neutrophil Extracellular Trap (NET)osis Formation and Reduced Phagocytosis of Polymorphonuclear Neutrophils

Spaceflight can dysregulate immunity, by way of increasing granulocytes numbers with impaired function. Polymorphonuclear neutrophils (PMN) are granulocytes that are first responders to infection or injury, and consist of the largest pool of immune cells in humans. PMNs function during innate immunity, through phagocytosis and promotion of inflammation, via the release of reactive oxygen species (ROS) mediators and granule-containing enzymes, such as myeloperoxidase (MPO) and NADPH oxidase-2 (NOX-2). In addition, neutrophil extracellular trap (NET) formation is another mechanism of PMN surveillance that works independently of engulfment phagocytosis, and is a last resort function that can induce NETosis or PMN-specific cell death. Previous studies in our lab have identified increased mature neutrophils, ROS and MPO production, and reduced phagocytosis in granulocytes in simulated microgravity (sug) models of hindlimb unloading (HU) in adult mice and leukocytes cultured in high-aspect rotating wall vessels (HARV-RWV). Since sug impaired phagocytosis, but improved enzymatic mediator production of MPO and redox molecules, we sought to address the third known function of PMNs, NETosis. For this, PMNs were culture in the presence or absence of the anti-oxidant N-acetyl cysteine (NAC), which rescued impaired phagocytosis that was present in sug without NAC treatment. Further, NETosis was induced in sug that was no different in the presence of NAC, suggesting NAC targets independent functions of PMNs under sug. Collectively, these results suggest modeled microgravity induced NETosis, which opens a new avenue for spaceflight studies in immune dysfunction.