Figure 7.
NADPH oxidase activity in primary phagocytic cells from PKCδ-deficient patients. (A) Neutrophil intracellular ROS production, as measured by DHR, upon PMA stimulation, for travel controls (ctrl; n = 11), PKCδ-deficient patients (n = 11), p40phox-deficient patients (n = 5), and gp91phox-deficient patients (n = 2), or upon E. coli stimulation, for travel controls (n = 5), PKCδ-deficient patients (n = 5), p40phox-deficient patients (n = 1), and gp91phox-deficient patients (n = 2). (B) Monocyte intracellular ROS production, measured by DHR, upon PMA stimulation, for travel controls (n = 11), PKCδ-deficient patients (n = 11), p40phox-deficient patients (n = 5), and gp91phox-deficient patients (n = 2), or upon E. coli stimulation, for travel controls (n = 5), PKCδ-deficient patients (n = 5), p40phox-deficient patients (n = 1), and gp91phox-deficient patients (n = 1). All values are expressed as a percentage of DHR oxidation normalized against travel controls. (C) Left: Quantification of NET formation for healthy controls (n = 6), PKCδ-deficient patients (n = 3), and a gp91phox-deficient patient, after PMA stimulation. All values are expressed as a percentage of NET-forming cells. Right: Representative images of PMA-induced NET formation by neutrophils of local and travel controls and a PKCδ-deficient patient (P17). Green represents myeloperoxidase, and blue, DNA (DAPI). Scale bar = 60 µm. (D) Extracellular H2O2 production in response to stimulation with IFN-γ, PMA, or both (mean ± SD) for MDMs from local controls (n = 10), travel controls (n = 10), PKCδ-deficient patients (n = 12), p40phox-deficient patients (n = 3), and gp91phox-deficient patients (n = 1). (E) Extracellular H2O2 production in response to stimulation with LPS, PMA, or both (mean ± SD), in MDDCs from local controls (n = 10), travel controls (n = 10), PKCδ-deficient patients (n = 8), p40phox-deficient patients (n = 2), and gp91phox-deficient patients (n = 2). (F) Western blot of MDMs of healthy controls (n = 4) and PKCδ-deficient patients (n = 5), without (–) or with (+) PMA stimulation. Solid bars between blots indicate different regions of the same membrane. In A–E, dots represent individual samples, and bars, the mean and SD. Two-way ANOVA was used in A–D; one-way ANOVA was used in C; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

NADPH oxidase activity in primary phagocytic cells from PKCδ-deficient patients. (A) Neutrophil intracellular ROS production, as measured by DHR, upon PMA stimulation, for travel controls (ctrl; n = 11), PKCδ-deficient patients (n = 11), p40phox-deficient patients (n = 5), and gp91phox-deficient patients (n = 2), or upon E. coli stimulation, for travel controls (n = 5), PKCδ-deficient patients (n = 5), p40phox-deficient patients (n = 1), and gp91phox-deficient patients (n = 2). (B) Monocyte intracellular ROS production, measured by DHR, upon PMA stimulation, for travel controls (n = 11), PKCδ-deficient patients (n = 11), p40phox-deficient patients (n = 5), and gp91phox-deficient patients (n = 2), or upon E. coli stimulation, for travel controls (n = 5), PKCδ-deficient patients (n = 5), p40phox-deficient patients (n = 1), and gp91phox-deficient patients (n = 1). All values are expressed as a percentage of DHR oxidation normalized against travel controls. (C) Left: Quantification of NET formation for healthy controls (n = 6), PKCδ-deficient patients (n = 3), and a gp91phox-deficient patient, after PMA stimulation. All values are expressed as a percentage of NET-forming cells. Right: Representative images of PMA-induced NET formation by neutrophils of local and travel controls and a PKCδ-deficient patient (P17). Green represents myeloperoxidase, and blue, DNA (DAPI). Scale bar = 60 µm. (D) Extracellular H2O2 production in response to stimulation with IFN-γ, PMA, or both (mean ± SD) for MDMs from local controls (n = 10), travel controls (n = 10), PKCδ-deficient patients (n = 12), p40phox-deficient patients (n = 3), and gp91phox-deficient patients (n = 1). (E) Extracellular H2O2 production in response to stimulation with LPS, PMA, or both (mean ± SD), in MDDCs from local controls (n = 10), travel controls (n = 10), PKCδ-deficient patients (n = 8), p40phox-deficient patients (n = 2), and gp91phox-deficient patients (n = 2). (F) Western blot of MDMs of healthy controls (n = 4) and PKCδ-deficient patients (n = 5), without (–) or with (+) PMA stimulation. Solid bars between blots indicate different regions of the same membrane. In A–E, dots represent individual samples, and bars, the mean and SD. Two-way ANOVA was used in A–D; one-way ANOVA was used in C; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

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