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Genetic disorders coupled to ROS deficiency.

O'Neill S, Brault J, Stasia MJ, Knaus UG - Redox Biol (2015)

Bottom Line: Disturbances such as continuously elevated ROS levels will result in oxidative stress and development of disease, but likewise, insufficient ROS production will be detrimental to health.More recently, additional diseases have been linked to functionally altered variants in genes encoding for other NADPH oxidases, such as for DUOX2/DUOXA2 in congenital hypothyroidism, or for the Nox2 complex, NOX1 and DUOX2 as risk factors for inflammatory bowel disease.A comprehensive overview of novel developments in terms of Nox/Duox-deficiency disorders is presented, combined with insights gained from structure-function studies that will aid in predicting functional defects of clinical variants.

View Article: PubMed Central - PubMed

Affiliation: Conway Institute, University College Dublin, Dublin, Ireland.

No MeSH data available.


Related in: MedlinePlus

Phenotypic and functional characterization of neutrophils and macrophages derived from WT and CGD iPSCs. (A) MGG staining showing the characteristic morphology of neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). (B) Electron microscopy shows the presence of cytoplasmic granules in neutrophils and vacuoles in macrophages (scale bar 2 µm). (C) NBT reduction assay on opsonized latex bead-activated WT or CGD iPSC-derived neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). ROS-mediated NBT reduction is shown as blue formazan precipitates in WT neutrophils and macrophages (black arrows). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
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f0020: Phenotypic and functional characterization of neutrophils and macrophages derived from WT and CGD iPSCs. (A) MGG staining showing the characteristic morphology of neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). (B) Electron microscopy shows the presence of cytoplasmic granules in neutrophils and vacuoles in macrophages (scale bar 2 µm). (C) NBT reduction assay on opsonized latex bead-activated WT or CGD iPSC-derived neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). ROS-mediated NBT reduction is shown as blue formazan precipitates in WT neutrophils and macrophages (black arrows). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Mentions: Recently, our group optimized protocols to differentiate iPSCs from X0-, AR470- and AR220-CGD patients' fibroblasts into neutrophils and macrophages [107]. Unlike other protocols described for CGD modeling, iPSCs were directly differentiated using adapted and optimized protocols from Choi et al. for the production of neutrophils and macrophages separately [108,109]. The average production of CD34+ progenitors was 1.5×106 cells after 10 days of differentiation of 10×106 iPSCs. Extensive characterization of CGD iPSC-derived neutrophils confirmed the presence of primary, secondary and tertiary granules in the cytoplasm (Fig. 4A, B). Cells were terminally differentiated into about 3×105 neutrophils or 3×107 macrophages in 25–28 days. CGD neutrophils and macrophages exhibited an oxidase-negative phenotype characterized by the absence of NADPH oxidase activity related to the absence of Nox2 and p22phox expression in X0-CGD and AR220-CGD cells (Fig. 4C). CGD iPSC-derived macrophages were able to phagocytose opsonized S. aureus or zymosan particles and produced pro- and anti-inflammatory cytokines after stimulation. iPSC-derived macrophages expressed classical CD14, CD45, and CD11b antigen surface markers and were HLA-DR−, CCR7−, and MR+, specific to the M2c subtype of macrophages, which are regulatory macrophages involved in immunosuppression and wound healing/tissue repair [107].


Genetic disorders coupled to ROS deficiency.

O'Neill S, Brault J, Stasia MJ, Knaus UG - Redox Biol (2015)

Phenotypic and functional characterization of neutrophils and macrophages derived from WT and CGD iPSCs. (A) MGG staining showing the characteristic morphology of neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). (B) Electron microscopy shows the presence of cytoplasmic granules in neutrophils and vacuoles in macrophages (scale bar 2 µm). (C) NBT reduction assay on opsonized latex bead-activated WT or CGD iPSC-derived neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). ROS-mediated NBT reduction is shown as blue formazan precipitates in WT neutrophils and macrophages (black arrows). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4550764&req=5

f0020: Phenotypic and functional characterization of neutrophils and macrophages derived from WT and CGD iPSCs. (A) MGG staining showing the characteristic morphology of neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). (B) Electron microscopy shows the presence of cytoplasmic granules in neutrophils and vacuoles in macrophages (scale bar 2 µm). (C) NBT reduction assay on opsonized latex bead-activated WT or CGD iPSC-derived neutrophils (upper panel, scale bar 10 µm) and macrophages (lower panel, scale bar 20 µm). ROS-mediated NBT reduction is shown as blue formazan precipitates in WT neutrophils and macrophages (black arrows). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Mentions: Recently, our group optimized protocols to differentiate iPSCs from X0-, AR470- and AR220-CGD patients' fibroblasts into neutrophils and macrophages [107]. Unlike other protocols described for CGD modeling, iPSCs were directly differentiated using adapted and optimized protocols from Choi et al. for the production of neutrophils and macrophages separately [108,109]. The average production of CD34+ progenitors was 1.5×106 cells after 10 days of differentiation of 10×106 iPSCs. Extensive characterization of CGD iPSC-derived neutrophils confirmed the presence of primary, secondary and tertiary granules in the cytoplasm (Fig. 4A, B). Cells were terminally differentiated into about 3×105 neutrophils or 3×107 macrophages in 25–28 days. CGD neutrophils and macrophages exhibited an oxidase-negative phenotype characterized by the absence of NADPH oxidase activity related to the absence of Nox2 and p22phox expression in X0-CGD and AR220-CGD cells (Fig. 4C). CGD iPSC-derived macrophages were able to phagocytose opsonized S. aureus or zymosan particles and produced pro- and anti-inflammatory cytokines after stimulation. iPSC-derived macrophages expressed classical CD14, CD45, and CD11b antigen surface markers and were HLA-DR−, CCR7−, and MR+, specific to the M2c subtype of macrophages, which are regulatory macrophages involved in immunosuppression and wound healing/tissue repair [107].

Bottom Line: Disturbances such as continuously elevated ROS levels will result in oxidative stress and development of disease, but likewise, insufficient ROS production will be detrimental to health.More recently, additional diseases have been linked to functionally altered variants in genes encoding for other NADPH oxidases, such as for DUOX2/DUOXA2 in congenital hypothyroidism, or for the Nox2 complex, NOX1 and DUOX2 as risk factors for inflammatory bowel disease.A comprehensive overview of novel developments in terms of Nox/Duox-deficiency disorders is presented, combined with insights gained from structure-function studies that will aid in predicting functional defects of clinical variants.

View Article: PubMed Central - PubMed

Affiliation: Conway Institute, University College Dublin, Dublin, Ireland.

No MeSH data available.


Related in: MedlinePlus