Poly(I:C) induced microRNA-146a regulates epithelial barrier and secretion of proinflammatory cytokines in human nasal epithelial cells

Background. Airway epithelium plays an important role during the development of allergic rhinitis (AR), which is characterized by production of thymic stromal lymphopoietin (TSLP), interleukin 33 (IL-33), and interleukin 25 (IL-25). IL-35, mainly expressed by Treg cells, have negative regulation in Th2, Th17, and ILC2 inflammation. However, the effect of IL-35 on human nasal epithelial cells (HNECs) especially the secretion of nasal epithelial-derived proinflammatory cytokines as well as the possible mechanism is still unclear. Methods. HNECs were cultured and stimulated by various stimulators. The expression of IL-33, IL-25, TSLP, eotaxin-1, eotaxin-2, and eotaxin-3 from supernatant was measured using real-time reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). AR mice were developed to verify the effect of IL-35 on nasal epithelial cells in vivo. Results. After Poly I:C stimulation, IL-35 inhibited the production of IL-25, and TSLP from HNECs increased significantly compared with baseline levels ( P < 0.05 ). After Dermatophagoides pteronyssinus or Aspergillus fumigatus stimulation, IL-35 inhibited the production of IL-25, IL-33, and TSLP from HNECs increased significantly compared with baseline levels ( P < 0.05 ). After Dermatophagoides pteronyssinus, IL-35 inhibited the production of eotaxin-1, eotaxin-2, and eotaxin-3 released from HNECs increased significantly compared with baseline levels ( P < 0.05 ). Similarly, IL-35-treated AR mice presented with decreased expression of IL-33, IL-25, TSLP, eotaxin-1, eotaxin-2, and eotaxin-3 in nasal lavage fluid. Conclusion. IL-35 suppressed both type 2 inflammation-inducing cytokines and eosinophil chemotactic factor from HNECs, suggesting the important role of IL-35 during the development of AR.

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The role of transcriptional factor p63 in regulation of epithelial barrier and ciliogenesis of human nasal epithelial cells

Scientific Reports ◽

10.1038/s41598-017-11481-w ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 13

Author(s):

Yakuto Kaneko ◽

Takayuki Kohno ◽

Takuya Kakuki ◽

Ken-ichi Takano ◽

Noriko Ogasawara ◽

...

Keyword(s):

Epithelial Cells ◽

Transcriptional Factor ◽

Epithelial Barrier ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells

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Urban Particulate Matters May Affect Endoplasmic Reticulum Stress and Tight Junction Disruption in Nasal Epithelial Cells

American Journal of Rhinology and Allergy ◽

10.1177/19458924211004006 ◽

2021 ◽

pp. 194589242110040

Author(s):

Soo Kyoung Park ◽

Sun Hee Yeon ◽

Mi-Ra Choi ◽

Seung Hyeon Choi ◽

Sung Bok Lee ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Epithelial Cells ◽

Er Stress ◽

Stress Responses ◽

Barrier Function ◽

Epithelial Barrier ◽

In Vivo Models ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells ◽

Mucosal Barrier Function

Background Exposure to airborne urban particulate matter (UPM) has been closely related to the development and aggravation of respiratory disease, including sinonasal disorders. Objective The aims of this study were to investigate the effect of UPM on nasal epithelial tight junctions (TJs) and mucosal barrier function and delineate the underlying mechanism by using both in vitro and in vivo models. Methods In this study, human nasal epithelial cells (hNECs) and BALB/c mice were exposed to UPMs. UPM 1648a and 1649 b were employed. TJ and endoplasmic reticulum (ER) stress marker expression was measured using western blot analysis and immunofluorescence. TJ integrity and nasal epithelial barrier function were evaluated by transepithelial electric resistance (TER) and paracellular flux. In addition, the effects of N‐acetyl‐L‐cysteine (NAC) on UPM-induced nasal epithelial cells were investigated. Results UPM significantly impaired the nasal epithelial barrier, as demonstrated by decreased protein expression of TJ and ER stress markers in human nasal epithelial cells. This finding was in parallel to reduced transepithelial electrical resistance and increased fluorescein isothiocyanate–dextran permeability. Pretreatment with NAC decreased the degree of UPM-mediated ER stress and restored nasal epithelial barrier disruption in human nasal epithelial cells (hNEC) and the nasal mucosa of experimental animals. Conclusion These data suggest that UPMs may induce nasal epithelial barrier dysfunction by targeting TJs and ER stress could be related in this process. Based on these results, we suggest that suppression of this process with an inhibitor targeting ER stress responses could represent a novel promising therapeutic target in UPM-induced sinonasal disease.

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Effects of HMGB1 on Tricellular Tight Junctions via TGF-β Signaling in Human Nasal Epithelial Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22168390 ◽

2021 ◽

Vol 22 (16) ◽

pp. 8390

Author(s):

Kizuku Ohwada ◽

Takumi Konno ◽

Takayuki Kohno ◽

Masaya Nakano ◽

Tsuyoshi Ohkuni ◽

...

Keyword(s):

Epithelial Cells ◽

Tight Junctions ◽

Chronic Rhinosinusitis ◽

Kinase Inhibitor ◽

High Mobility ◽

Epithelial Barrier ◽

Type I ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells ◽

Matrigel Culture

The airway epithelium of the human nasal mucosa acts as a physical barrier that protects against inhaled substances and pathogens via bicellular and tricellular tight junctions (bTJs and tTJs) including claudins, angulin-1/LSR and tricellulin. High mobility group box-1 (HMGB1) increased by TGF-β1 is involved in the induction of nasal inflammation and injury in patients with allergic rhinitis, chronic rhinosinusitis, and eosinophilic chronic rhinosinusitis. However, the detailed mechanisms by which this occurs remain unknown. In the present study, to investigate how HMGB1 affects the barrier of normal human nasal epithelial cells, 2D and 2.5D Matrigel culture of primary cultured human nasal epithelial cells were pretreated with TGF-β type I receptor kinase inhibitor EW-7197 before treatment with HMGB1. Knockdown of angulin-1/LSR downregulated the epithelial barrier. Treatment with EW-7197 decreased angulin-1/LSR and concentrated the expression at tTJs from bTJs and increased the epithelial barrier. Treatment with a binder to angulin-1/LSR angubindin-1 decreased angulin-1/LSR and the epithelial barrier. Treatment with HMGB1 decreased angulin-1/LSR and the epithelial barrier. In 2.5D Matrigel culture, treatment with HMGB1 induced permeability of FITC-dextran (FD-4) into the lumen. Pretreatment with EW-7197 prevented the effects of HMGB1. HMGB1 disrupted the angulin-1/LSR-dependent epithelial permeability barriers of HNECs via TGF-β signaling in HNECs.

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Fluticasone Propionate Suppresses Poly(I:C)-Induced ACE2 in Primary Human Nasal Epithelial Cells

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.655666 ◽

2021 ◽

Vol 11 ◽

Author(s):

Akira Nakazono ◽

Yuji Nakamaru ◽

Mahnaz Ramezanpour ◽

Takeshi Kondo ◽

Masashi Watanabe ◽

...

Keyword(s):

Epithelial Cells ◽

Fluticasone Propionate ◽

Nasal Mucosa ◽

Fold Change ◽

Innate Immune ◽

Cell Entry ◽

Poly I:C ◽

Nasal Epithelial Cells ◽

Tlr Agonists ◽

Human Nasal Epithelial Cells

BackgroundFrom the first detection in 2019, SARS-CoV-2 infections have spread rapidly worldwide and have been proven to cause an urgent and important health problem. SARS-CoV-2 cell entry depends on two proteins present on the surface of host cells, angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2). The nasal cavity is thought to be one of the initial sites of infection and a possible reservoir for dissemination within and between individuals. However, it is not known how the expression of these genes is regulated in the nasal mucosa.ObjectiveIn this study, we examined whether the expression of ACE2 and TMPRSS2 is affected by innate immune signals in the nasal mucosa. We also investigated how fluticasone propionate (FP), a corticosteroid used as an intranasal steroid spray, affects the gene expression.MethodsPrimary human nasal epithelial cells (HNECs) were collected from the nasal mucosa and incubated with Toll-like receptor (TLR) agonists and/or fluticasone propionate (FP), followed by quantitative PCR, immunofluorescence, and immunoblot analyses.ResultsAmong the TLR agonists, the TLR3 agonist Poly(I:C) significantly increased ACE2 and TMPRSS2 mRNA expression in HNECs (ACE2 36.212±11.600-fold change, p<0.0001; TMPRSS2 5.598±2.434-fold change, p=0.031). The ACE2 protein level was also increased with Poly(I:C) stimulation (2.884±0.505-fold change, p=0.003). The Poly(I:C)-induced ACE2 expression was suppressed by co-incubation with FP (0.405±0.312-fold change, p=0.044).ConclusionThe activation of innate immune signals via TLR3 promotes the expression of genes related to SARS-CoV2 cell entry in the nasal mucosa, although this expression is suppressed in the presence of FP. Further studies are required to evaluate whether FP suppresses SARS-CoV-2 viral cell entry.

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Poly(I:C) reduces expression of JAM-A and induces secretion of IL-8 and TNF-α via distinct NF-κB pathways in human nasal epithelial cells

Toxicology and Applied Pharmacology ◽

10.1016/j.taap.2010.09.023 ◽

2011 ◽

Vol 250 (1) ◽

pp. 29-38 ◽

Cited By ~ 32

Author(s):

Tsuyoshi Ohkuni ◽

Takashi Kojima ◽

Noriko Ogasawara ◽

Tomoyuki Masaki ◽

Jun Fuchimoto ◽

...

Keyword(s):

Epithelial Cells ◽

Poly I:C ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells ◽

Tnf Α

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Primary human nasal epithelial cells: a source of poly (I:C) LMW-induced IL-6 production

Scientific Reports ◽

10.1038/s41598-018-29765-0 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 7

Author(s):

Mahnaz Ramezanpour ◽

Harrison Bolt ◽

Alkis James Psaltis ◽

Peter-John Wormald ◽

Sarah Vreugde

Keyword(s):

Epithelial Cells ◽

Poly I:C ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells

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Elovanoids Counteract Inflammatory Signaling, Autophagy, Endoplasmic Reticulum Stress, and Senescence Gene Programming in Human Nasal Epithelial Cells Exposed to Allergens

Pharmaceutics ◽

10.3390/pharmaceutics14010113 ◽

2022 ◽

Vol 14 (1) ◽

pp. 113

Author(s):

Alfredo Resano ◽

Surjyadipta Bhattacharjee ◽

Miguel Barajas ◽

Khanh V. Do ◽

Roberto Aguado-Jiménez ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Epithelial Cells ◽

Inflammatory Responses ◽

House Dust Mites ◽

Poly I:C ◽

Lipid Mediator ◽

Omega 3 ◽

Nasal Epithelial Cells ◽

Human Nasal Epithelial Cells

To contribute to further understanding the cellular and molecular complexities of inflammatory-immune responses in allergic disorders, we have tested the pro-homeostatic elovanoids (ELV) in human nasal epithelial cells (HNEpC) in culture challenged by several allergens. ELV are novel bioactive lipid mediators synthesized from the omega-3 very-long-chain polyunsaturated fatty acids (VLC-PUFA,n-3). We ask if: (a) several critical signaling events that sustain the integrity of the human nasal epithelium and other organ barriers are perturbed by house dust mites (HDM) and other allergens, and (b) if ELV would participate in beneficially modulating these events. HDM is a prevalent indoor allergen that frequently causes allergic respiratory diseases, including allergic rhinitis and allergic asthma, in HDM-sensitized individuals. Our study used HNEpC as an in vitro model to study the effects of ELV in counteracting HDM sensitization resulting in inflammation, endoplasmic reticulum (ER) stress, autophagy, and senescence. HNEpC were challenged with the following allergy inducers: LPS, poly(I:C), or Dermatophagoides farinae plus Dermatophagoides pteronyssinus extract (HDM) (30 µg/mL), with either phosphate-buffered saline (PBS) (vehicle) or ELVN-34 (500 nM). Results show that ELVN-34 promotes cell viability and reduces cytotoxicity upon HDM sensitization of HNEpC. This lipid mediator remarkably reduces the abundance of pro-inflammatory cytokines and chemokines IL-1β, IL-8, VEGF, IL-6, CXCL1, CCL2, and cell adhesion molecule ICAM1 and restores the levels of the pleiotropic anti-inflammatory IL-10. ELVN-34 also lessens the expression of senescence gene programming as well as of gene transcription engaged in pro-inflammatory responses. Our data also uncovered that HDM triggered the expression of key genes that drive autophagy, unfolded protein response (UPR), and matrix metalloproteinases (MMP). ELVN-34 has been shown to counteract these effects effectively. Together, our data reveal a novel, pro-homeostatic, cell-protective lipid-signaling mechanism in HNEpC as potential therapeutic targets for allergies.

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