Intermittent Hypoxia Affects the Spontaneous Differentiation In Vitro of Human Neutrophils into Long-Lived Giant Phagocytes

Figure 8

Expression of NADPH oxidase subunits in giant phagocytes (Gϕ) and the effects of NAC on their expression. Freshly isolated PMN were exposed for 24 h to intermittent hypoxia (IH, 56 cycles), sustained hypoxia (SH), or normoxia (N) and then cultured at normoxia for additional six days. NAC (20 μM) was added to PMN cultures 10 min prior to exposing to N, IH, or SH. Equal volumes of DMSO were added as a negative control. Cytospins were prepared and analyzed by confocal microscopy (see Materials and Methods). The developed Gϕ were stained by double immunofluorescence: (a) for CD66b (red) and gp91-phox (green) in untreated and NAC-treated Gϕ and (b) for gp91-phox (green) and p22-phox (red) in untreated and NAC-treated Gϕ. Nuclei were stained with DAPI. Representative photomicrographs out of 3 independent experiments.

(a)

(b)