Influenza virus M2 protein inhibits epithelial sodium channels by increasing reactive oxygen species

Lazrak, Ahmed; Iles, Karen E.; Liu, Gang; Noah, Diana L.; Noah, James W.; Matalon, Sadis

doi:10.1096/fj.09-135590

Cited by 84 publications

(76 citation statements)

References 61 publications

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“…Infection with H5N1, compared with seasonal H1N1, influenza virus also led to greater down-regulation of the Na,K-ATPase and CFTR transporter proteins in alveolar epithelium. Influenza virus infection was previously shown to impair alveolar fluid transport by inhibiting epithelial sodium channel activity through interaction of viral hemagglutinin (15) and matrix (16) proteins with the epithelial sodium channel. To our knowledge, ours is the first demonstration of impaired alveolar fluid clearance and protein permeability due to soluble mediators released by infected alveolar epithelium.…”

Section: Discussionmentioning

confidence: 99%

Human mesenchymal stromal cells reduce influenza A H5N1-associated acute lung injury in vitro and in vivo

Chan

Kuok

Leung

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

182

181

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Influenza can cause acute lung injury. Because immune responses often play a role, antivirals may not ensure a successful outcome. To identify pathogenic mechanisms and potential adjunctive therapeutic options, we compared the extent to which avian influenza A/H5N1 virus and seasonal influenza A/H1N1 virus impair alveolar fluid clearance and protein permeability in an in vitro model of acute lung injury, defined the role of virus-induced soluble mediators in these injury effects, and demonstrated that the effects are prevented or reduced by bone marrow-derived multipotent mesenchymal stromal cells. We verified the in vivo relevance of these findings in mice experimentally infected with influenza A/H5N1. We found that, in vitro, the alveolar epithelium’s protein permeability and fluid clearance were dysregulated by soluble immune mediators released upon infection with avian (A/Hong Kong/483/97, H5N1) but not seasonal (A/Hong Kong/54/98, H1N1) influenza virus. The reduced alveolar fluid transport associated with down-regulation of sodium and chloride transporters was prevented or reduced by coculture with mesenchymal stromal cells. In vivo, treatment of aged H5N1-infected mice with mesenchymal stromal cells increased their likelihood of survival. We conclude that mesenchymal stromal cells significantly reduce the impairment of alveolar fluid clearance induced by A/H5N1 infection in vitro and prevent or reduce A/H5N1-associated acute lung injury in vivo. This potential adjunctive therapy for severe influenza-induced lung disease warrants rapid clinical investigation.

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Section: Discussionmentioning

confidence: 99%

Human mesenchymal stromal cells reduce influenza A H5N1-associated acute lung injury in vitro and in vivo

Chan

Kuok

Leung

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

182

181

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“…Voltage Clamp Recordings of Whole-cell ENaC CurrentsWhole-cell cation currents were recorded 48 -72 h post-injection across the entire oocyte membrane using the two-electrode voltage clamp technique (35). Briefly, the oocytes were held in a small groove in a chamber of 1-ml volume at room temperature (21°C).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of Lung Fluid Clearance and Epithelial Na+ Channels by Chlorine, Hypochlorous Acid, and Chloramines

Song¹,

Wei²,

Zhou³

et al. 2010

Journal of Biological Chemistry

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We investigated the mechanisms by which chlorine (Cl 2 ) and its reactive byproducts inhibit Na ؉ -dependent alveolar fluid clearance (AFC) in vivo and the activity of amiloridesensitive epithelial Na ؉ channels (ENaC) by measuring AFC in mice exposed to Cl 2 (0 -500 ppm for 30 min) and Na ؉ and amiloride-sensitive currents (I Na and I amil , respectively) across Xenopus oocytes expressing human ␣-, ␤-, and ␥-ENaC incubated with HOCl (1-2000 M). Both Cl 2 and HOCl-derived products decreased AFC in mice and whole cell and single channel I Na in a dose-dependent manner; these effects were counteracted by serine proteases. Mass spectrometry analysis of the oocyte recording medium identified organic chloramines formed by the interaction of HOCl with HEPES (used as an extracellular buffer). In addition, chloramines formed by the interaction of HOCl with taurine or glycine decreased I Na in a similar fashion. Preincubation of oocytes with serine proteases prevented the decrease of I Na by HOCl, whereas perfusion of oocytes with a synthetic 51-mer peptide corresponding to the putative furin and plasmin cleaving segment in the ␥-ENaC subunit restored the ability of HOCl to inhibit I Na . Finally, I Na of oocytes expressing wild type ␣-and ␥-ENaC and a mutant form of ␤ENaC (S520K), known to result in ENaC channels locked in the open position, were not altered by HOCl. We concluded that HOCl and its reactive intermediates (such as organic chloramines) inhibit ENaC by affecting channel gating, which could be relieved by proteases cleavage.The balance of fluid covering the respiratory and alveolar epithelia is determined in part by the ability of these cells to transport sodium (Na ϩ ) and chloride (Cl Ϫ ) ions in a vectorial fashion. Active Na ϩ reabsorption across lung epithelia requires the coordinated entry of Na ϩ ions through cation-and Na ϩ -selective amiloride-sensitive channels (ENaC) 5 located at the apical membranes, their extrusion across the basolateral membranes by the electrogenic Na ϩ -K ϩ -ATPase, and the passive movement of K ϩ ions through basolateral K ϩ channels. The entry of Na ϩ ions through apical pathways is thought to be the rate-limiting step in this process (1-3). To preserve neutrality, Cl Ϫ ions follow Na ϩ ions both through transcellular and paracellular pathways (4, 5). The coordinated movement of Na ϩ and Cl Ϫ ions creates an oncotic gradient favoring the absorption of alveolar fluid.Injury to either apical or basolateral pathways by partially reduced intermediates may lead to impairment of fluid reabsorption, which in turn may result in pulmonary edema, hypoxemia, and eventually death from respiratory failure (6 -9). One such specie is hypochlorous acid (HOCl) 6 , which may be generated either endogenously or exogenously. Millimolar concentrations of HOCl may be generated by activated neutrophils and eosinophils by the catalytic actions of neutrophil-and eosinophil-derived myeloperoxidases on chloride (Cl Ϫ ) and hydrogen peroxide (H 2 O 2 ) in close proximity of the apical and basolatera...

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“…Single-channel activity in both ATII and ATI cells was recorded using the cellattached mode of the patch clamp technique (19,20). ATII cells were identified by the presence of scattered green fluorescence after incubation with Lysotracker Green (catalogue number DND-26; Invitrogen, Eugene, OR).…”

Section: Recordings From Mouse Lungs Of Enac Single-channel Activitymentioning

confidence: 99%

Regulation of Alveolar Epithelial Na⁺ Channels by ERK1/2 in Chlorine-Breathing Mice

Lazrak¹,

Chen²,

Jurkuvenaite³

et al. 2012

Am J Respir Cell Mol Biol

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The mechanisms by which the exposure of mice to Cl 2 decreases vectorial Na 1 transport and fluid clearance across their distal lung spaces have not been elucidated. We examined the biophysical, biochemical, and physiological changes of rodent lung epithelial Na 1 channels (ENaCs) after exposure to Cl 2 , and identified the mechanisms involved. We measured amiloride-sensitive short-circuit currents (I amil ) across isolated alveolar Type II (ATII) cell monolayers and ENaC single-channel properties by patching ATII and ATI cells in situ. a-ENaC, g-ENaC, total and phosphorylated extracellular signalrelated kinase (ERK)1/2, and advanced products of lipid peroxidation in ATII cells were measured by Western blot analysis. Concentrations of reactive intermediates were assessed by electron spin resonance (ESR). Amiloride-sensitive Na 1 channels with conductances of 4.5 and 18 pS were evident in ATI and ATII cells in situ of air-breathing mice. At 1 hour and 24 hours after exposure to Cl 2 , the open probabilities of these two channels decreased. This effect was prevented by incubating lung slices with inhibitors of ERK1/2 or of proteasomes and lysosomes. The exposure of ATII cell monolayers to Cl 2 increased concentrations of reactive intermediates, leading to ERK1/2 phosphorylation and decreased I amil and a-ENaC concentrations at 1 hour and 24 hours after exposure. The administration of antioxidants to ATII cells before and after exposure to Cl 2 decreased concentrations of reactive intermediates and ERK1/2 activation, which mitigated the decrease in I amil and ENaC concentrations. The reactive intermediates formed during and after exposure to Cl 2 activated ERK1/2 in ATII cells in vitro and in vivo, leading to decreased ENaC concentrations and activity.

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Influenza virus M2 protein inhibits epithelial sodium channels by increasing reactive oxygen species

Cited by 84 publications

References 61 publications

Human mesenchymal stromal cells reduce influenza A H5N1-associated acute lung injury in vitro and in vivo

Human mesenchymal stromal cells reduce influenza A H5N1-associated acute lung injury in vitro and in vivo

Inhibition of Lung Fluid Clearance and Epithelial Na+ Channels by Chlorine, Hypochlorous Acid, and Chloramines

Regulation of Alveolar Epithelial Na⁺ Channels by ERK1/2 in Chlorine-Breathing Mice

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Influenza virus M2 protein inhibits epithelial sodium channels by increasing reactive oxygen species

Cited by 84 publications

References 61 publications

Human mesenchymal stromal cells reduce influenza A H5N1-associated acute lung injury in vitro and in vivo

Human mesenchymal stromal cells reduce influenza A H5N1-associated acute lung injury in vitro and in vivo

Inhibition of Lung Fluid Clearance and Epithelial Na+ Channels by Chlorine, Hypochlorous Acid, and Chloramines

Regulation of Alveolar Epithelial Na+ Channels by ERK1/2 in Chlorine-Breathing Mice

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Regulation of Alveolar Epithelial Na⁺ Channels by ERK1/2 in Chlorine-Breathing Mice