The X-CGD PLB-985 Cell Model for NOX2 Structure-Function Analysis

Beaumel, Sylvain; Stasia, Marie José

doi:10.1007/978-1-4939-9424-3_10

Cited by 5 publications

(5 citation statements)

References 26 publications

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“…Analysis of X-CGD-related mutations revealed the existence of three distinct cases defined by a total absence of NOX2 (X 0 -CGD), or by a low expression of the mutated protein correlated with a reduced oxidase activity (X minus-CGD), or finally by normal expression of NOX2 but a loss of oxidase activity (X + CGD) [345]. These alternatives provided crucial models that led to definitions of functional domains and residues in NOX2 [346]. X + CGD-related mutations mainly affect the catalytic activity of the oxidase, while the mutations responsible for X minus CGD appear to affect the proper maturation and correct folding of NOX2 [347].…”

Section: Chronic Granulomatous Diseasementioning

confidence: 99%

NADPH Oxidases (NOX): An Overview from Discovery, Molecular Mechanisms to Physiology and Pathology

et al. 2021

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The reactive oxygen species (ROS)-producing enzyme NADPH oxidase (NOX) was first identified in the membrane of phagocytic cells. For many years, its only known role was in immune defense, where its ROS production leads to the destruction of pathogens by the immune cells. NOX from phagocytes catalyzes, via one-electron trans-membrane transfer to molecular oxygen, the production of the superoxide anion. Over the years, six human homologs of the catalytic subunit of the phagocyte NADPH oxidase were found: NOX1, NOX3, NOX4, NOX5, DUOX1, and DUOX2. Together with the NOX2/gp91phox component present in the phagocyte NADPH oxidase assembly itself, the homologs are now referred to as the NOX family of NADPH oxidases. NOX are complex multidomain proteins with varying requirements for assembly with combinations of other proteins for activity. The recent structural insights acquired on both prokaryotic and eukaryotic NOX open new perspectives for the understanding of the molecular mechanisms inherent to NOX regulation and ROS production (superoxide or hydrogen peroxide). This new structural information will certainly inform new investigations of human disease. As specialized ROS producers, NOX enzymes participate in numerous crucial physiological processes, including host defense, the post-translational processing of proteins, cellular signaling, regulation of gene expression, and cell differentiation. These diversities of physiological context will be discussed in this review. We also discuss NOX misregulation, which can contribute to a wide range of severe pathologies, such as atherosclerosis, hypertension, diabetic nephropathy, lung fibrosis, cancer, or neurodegenerative diseases, giving this family of membrane proteins a strong therapeutic interest.

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Section: Chronic Granulomatous Diseasementioning

confidence: 99%

NADPH Oxidases (NOX): An Overview from Discovery, Molecular Mechanisms to Physiology and Pathology

et al. 2021

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“…It was shown that differentiated PLB-985 cells expressed NADPH oxidase components [ 17 , 18 ]. Antibody (7D5 monoclonal antibody) directed against the extracellular loop of gp91 phox in PLB-985 revealed that cyt b 558 should be present at the plasma membrane [ 19 ]. However, the proportion of gp91 phox at the plasma membrane or in the intracellular compartment is unknown.…”

Section: Discussionmentioning

confidence: 99%

Accumulation of Cytochrome b558 at the Plasma Membrane: Hallmark of Oxidative Stress in Phagocytic Cells

Owusu

Dupré-Crochet

Bizouarn

et al. 2022

IJMS

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Neutrophils play a very key role in the human immune defense against pathogenic infections. The predominant players in this role during the activation of neutrophils are the release of cytotoxic agents stored in the granules and secretory vesicles and the massive production of reactive oxygen species (ROS) initiated by the enzyme NADPH oxidase. In addition, in living organisms, cells are continuously exposed to endogenous (inflammations, elevated neutrophil presence in the vicinity) and exogenous ROS at low and moderate levels (travels by plane, radiotherapy, space irradiation, blood banking, etc.). To study these effects, we used ROS induced by gamma radiation from low (0.2 Gy) to high (25 Gy) dose levels on PLB-985 cells from a myeloid cell line differentiated to neutrophil-like cells that are considered a good alternative to neutrophils. We determined a much longer lifetime of PLB-985 cells than that of neutrophils, which, as expected, decreased by increasing the irradiation dose. In the absence of any secondary stimulus, a very low production of ROS is detected with no significant difference between irradiated and non-irradiated cells. However, in phagocytosing cells, irradiation doses above 2 Gy enhanced oxidative burst in PLB-985 cells. Whatever the irradiation dose, NADPH oxidase devoid of its cytosolic regulatory units is observed at the plasma membrane in irradiated PLB-985 cells. This result is different from that observed for irradiated neutrophils in which irradiation also induced a translocation of regulatory subunits suggesting that the signal transduction mechanism or pathway operate differently in both cells.

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“…However, evaluating these new therapies requires cellular models. The first described in 1993 [50], and the only cell-based model mimicking the X-CGD form, available for this purpose, was the knockout CYBB PLB-985 cell [50,51]. These cells are bi-potential and can differentiate into either granulocytic or monocytic forms.…”

Section: Nox2 Deficiency: Cellular Modelsmentioning

confidence: 99%

“…In these cells, the X chromosome-linked gp91 phox gene was disrupted by homologous recombination to generate cells that, after differentiation to granulocytes, did not generate O 2 − , reproducing the phenotype of the X 0 -CGD patients with this mutation [50]. A second method to generate mutated NOX2 PLB-985 cells was described by Beaumel et al [51]. PLB-985 cells were transfected with various NOX2 mutations, cultured, and differentiated into neutrophils or monocytes/macrophages to evaluate the impact of these mutations and to study the relationships between NOX2 structure function [51].…”

Section: Nox2 Deficiency: Cellular Modelsmentioning

confidence: 99%

“…A second method to generate mutated NOX2 PLB-985 cells was described by Beaumel et al [51]. PLB-985 cells were transfected with various NOX2 mutations, cultured, and differentiated into neutrophils or monocytes/macrophages to evaluate the impact of these mutations and to study the relationships between NOX2 structure function [51].…”

Section: Nox2 Deficiency: Cellular Modelsmentioning

confidence: 99%

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Structure, Activation, and Regulation of NOX2: At the Crossroad between the Innate Immunity and Oxidative Stress-Mediated Pathologies

et al. 2023

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Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) is a multisubunit enzyme complex that participates in the generation of superoxide or hydrogen peroxide (H2O2) and plays a key role in several biological functions. Among seven known NOX isoforms, NOX2 was the first identified in phagocytes but is also expressed in several other cell types including endothelial cells, platelets, microglia, neurons, and muscle cells. NOX2 has been assigned multiple roles in regulating many aspects of innate and adaptive immunity, and human and mouse models of NOX2 genetic deletion highlighted this key role. On the other side, NOX2 hyperactivation is involved in the pathogenesis of several diseases with different etiologies but all are characterized by an increase in oxidative stress and inflammatory process. From this point of view, the modulation of NOX2 represents an important therapeutic strategy aimed at reducing the damage associated with its hyperactivation. Although pharmacological strategies to selectively modulate NOX2 are implemented thanks to new biotechnologies, this field of research remains to be explored. Therefore, in this review, we analyzed the role of NOX2 at the crossroads between immunity and pathologies mediated by its hyperactivation. We described (1) the mechanisms of activation and regulation, (2) human, mouse, and cellular models studied to understand the role of NOX2 as an enzyme of innate immunity, (3) some of the pathologies associated with its hyperactivation, and (4) the inhibitory strategies, with reference to the most recent discoveries.

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The X-CGD PLB-985 Cell Model for NOX2 Structure-Function Analysis

Cited by 5 publications

References 26 publications

NADPH Oxidases (NOX): An Overview from Discovery, Molecular Mechanisms to Physiology and Pathology

NADPH Oxidases (NOX): An Overview from Discovery, Molecular Mechanisms to Physiology and Pathology

Accumulation of Cytochrome b558 at the Plasma Membrane: Hallmark of Oxidative Stress in Phagocytic Cells

Structure, Activation, and Regulation of NOX2: At the Crossroad between the Innate Immunity and Oxidative Stress-Mediated Pathologies

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