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Previously we identified, for the first time, a new small-size subset of neutrophil-derived giant phagocytes (G ϕ ) which spontaneously develop  in vitro  without additional growth factors or cytokines. G ϕ  are CD66b + /CD63 + /MPO + /LC3B +  and are characterized by extended lifespan, large phagolysosomes, active phagocytosis, and reactive oxygen species (ROS) production, and autophagy largely controls their formation. Hypoxia, and particularly hypoxia/reoxygenation, is a prominent feature of many pathological processes. Herein we investigated G ϕ  formation by applying various hypoxic conditions. Chronic intermittent hypoxia (IH) (29 cycles/day for 5 days) completely abolished G ϕ  formation, while acute IH had dose-dependent effects. Exposure to 24 h (56 IH cycles) decreased their size, yield, phagocytic ability, autophagy, mitophagy, and gp91- phox /p22- phox  expression, whereas under 24 h sustained hypoxia (SH) the size and expression of LC3B and gp91- phox /p22- phox  resembled G ϕ  formed in normoxia. Diphenyl iodide (DPI), a NADPH oxidase inhibitor, as well as the PI3K/Akt and autophagy inhibitor LY294002 abolished G ϕ  formation at all oxygen conditions. However, the potent antioxidant, N-acetylcysteine (NAC) abrogated the effects of IH by inducing large CD66b + /LC3B +  G ϕ  and increased both NADPH oxidase expression and phagocytosis. These findings suggest that NADPH oxidase, autophagy, and the PI3K/Akt pathway are involved in G ϕ  development.

Intermittent Hypoxia Affects the Spontaneous DifferentiationIn Vitroof Human Neutrophils into Long-Lived Giant Phagocytes