The proton ATPase inhibitor bafilomycin A1 reduces the release of rhinovirus C and cytokines from primary cultures of human nasal epithelial cells

Polycationic proteins alter electrolyte transport by epithelium and endothelium, and in asthma are thought to disrupt the airway epithelium and contribute to hyperresponsiveness and airway plugging. In the present study, we used primary cultures of human nasal epithelial cells to investigate the response of respiratory tract epithelium to luminal presentation of a polycationic protein, protamine. Protamine (100 micrograms/ml) in the apical bathing solution had no significant effect on basal transepithelial resistance (Rt) but decreased short-circuit current (Isc) and hyperpolarized the apical membrane, indicating that Na+ absorption had been inhibited. Pretreating with amiloride inverted the response to protamine, resulting in an increase in Isc, depolarization of the apical membrane, and decrease in the fractional resistance of the apical membrane (fRa). The increase in Isc was inhibited by pretreatment with bumetanide. These results indicated that protamine augmented amiloride-induced Cl- secretion. Induction of Cl- secretion by bathing the apical surface in 3 mM Cl(-)-Ringer solution similarly resulted in protamine-induced depolarization of the apical membrane. Heparin precipitated protamine from solution and reversed the Isc responses. In summary, low concentrations of polycationic protein can alter electrolyte transport by human airway epithelium without desquamation, and the response is dependent on the secretory state of the tissue.

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Expression Of Surfactant Protein A In Human Nasal Epithelial Cells After Infection With Rhinovirus C In The Presence of Histamine

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2016.12.255 ◽

2017 ◽

Vol 139 (2) ◽

pp. AB64

Author(s):

Georgios T. Noutsios ◽

Erin Romero ◽

Amanda L. Willis ◽

Jaeden T. Calton ◽

Julie G. Ledford ◽

...

Keyword(s):

Epithelial Cells ◽

Protein A ◽

Surfactant Protein ◽

Surfactant Protein A ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells ◽

Rhinovirus C

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Dual mechanisms for Na-K-Cl cotransport regulation in airway epithelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.1993.264.1.c189 ◽

1993 ◽

Vol 264 (1) ◽

pp. C189-C200 ◽

Cited By ~ 32

Author(s):

M. Haas ◽

D. G. McBrayer ◽

J. R. Yankaskas

Keyword(s):

Epithelial Cells ◽

Primary Cultures ◽

Airway Epithelial Cells ◽

Beta Agonist ◽

Secondary Response ◽

Nasal Epithelial Cells ◽

Cl Channel ◽

Human Nasal Epithelial Cells ◽

Stimulation Of ◽

Airway Cells

To investigate cellular mechanisms involved in the regulation of basolateral Na-K-Cl cotransport in airway epithelia, we determined saturable basolateral [3H]bumetanide binding, a measure of functioning cotransporters, in primary cultures of canine tracheal and human nasal epithelial cells, including cells from patients with cystic fibrosis (CF). As we previously reported [M. Haas, L. G. Johnson, and R. C. Boucher. Am. J. Physiol. 259 (Cell Physiol. 28): C557-C569, 1990], isoproterenol and hypertonic cell shrinkage produce an equivalent stimulation of [3H]bumetanide binding to dog tracheal cells. We now find that apical ATP and UTP, which stimulate apical Cl channels and Cl secretion in normal and CF airway cells by an adenosine 3',5'-cyclic monophosphate (cAMP)-independent mechanism (S. J. Mason, A. M. Paradiso, and R. C. Boucher. Br. J. Pharmacol. 103: 1649-1656, 1991), increase basolateral [3H]bumetanide binding to dog tracheal cells to the same extent as do isoproterenol and hypertonic shrinkage. The stimulatory effects of ATP and UTP on binding are inhibited by apical addition of a Cl channel blocker, the indanyloxyacetic acid derivative IAA-94 (0.2 mM), or by raising basolateral K concentration ([K]b) from 3.3 to 40 mM, suggesting these effects are secondary to apical Cl efflux via channels. Apical IAA-94 and increased [K]b also inhibit stimulation of binding by isoproterenol by approximately 50%, suggesting that part (but not all) of the effect of the beta-agonist on basolateral cotransport is secondary to apical Cl efflux, with an additional component of direct stimulation of cotransport via cAMP. In support of this interpretation, we find that isoproterenol and a membrane-permeable cAMP analogue increase [3H]bumetanide binding to primary cultures of CF nasal epithelial cells, in which significant cAMP-mediated stimulation of apical Cl efflux does not occur. [3H]bumetanide binding to CF nasal cells is also stimulated by apical ATP, and levels of saturable [3H]bumetanide binding to CF cells are 1.3-1.5 times those in non-CF nasal cells under both basal and stimulated conditions. The results suggest that basolateral Na-K-Cl cotransport in airway cells may be upregulated in two distinct ways: 1) directly via a cAMP-dependent cascade, and 2) as a secondary response to apical Cl channel activation. Both of these mechanisms appear to be intact in CF.

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Ginsenoside Re prevents disruption of tight junction induced by rhinovirus through inhibition of ROS-mediated phosphatases inactivation in human nasal epithelial cells

10.26226/morressier.5acdc65ed462b8029238de02 ◽

2018 ◽

Author(s):

Seon-Tae Kim

Keyword(s):

Epithelial Cells ◽

Tight Junction ◽

Ginsenoside Re ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells

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miR-1287-5p upregulation inhibits the EMT and pro-inflammatory cytokines in LPS-induced human nasal epithelial cells (HNECs)

Transplant Immunology ◽

10.1016/j.trim.2021.101429 ◽

2021 ◽

pp. 101429

Author(s):

Wenwei Hao ◽

Yongping Zhu ◽

Ying Guo ◽

Haowei Wang

Keyword(s):

Epithelial Cells ◽

Inflammatory Cytokines ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells ◽

Pro Inflammatory Cytokines

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Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro

Viruses ◽

10.3390/v13020282 ◽

2021 ◽

Vol 13 (2) ◽

pp. 282

Author(s):

Finny S. Varghese ◽

Esther van Woudenbergh ◽

Gijs J. Overheul ◽

Marc J. Eleveld ◽

Lisa Kurver ◽

...

Keyword(s):

Epithelial Cells ◽

Early Stage ◽

Antiviral Drugs ◽

Target Cells ◽

Nasal Epithelial Cells ◽

Cytokines And Chemokines ◽

Human Nasal Epithelial Cells ◽

Apoptotic Protein ◽

Air Liquid Interface

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and it has infected over 100 million people in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures, including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, which was originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but it strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, which is in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells that were cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of specific sets of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.

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