The expression of the eotaxins IL-6 and CXCL8 in human epithelial cells from various levels of the respiratory tract

AbstractAirway epithelium acts as multifunctional site of response in the respiratory tract. Epithelial activity plays an important part in the pathophysiology of obstructive lung disease. In this study, we compare normal human epithelial cells from various levels of the respiratory tract in terms of their reactivity to pro-allergic and pro-inflammatory stimulation. Normal human nasal, bronchial and small airway epithelial cells were stimulated with IL-4 and IL-13. The expressions of the eotaxins IL-6 and CXCL8 were evaluated at the mRNA and protein levels. The effects of pre-treatment with IFN-γ on the cell reactivity were measured, and the responses to TNF-α, LPS and IFN-γ were evaluated. All of the studied primary cells expressed CCL26, IL-6 and IL-8 after IL-4 or IL-13 stimulation. IFN-γ pre-treatment resulted in decreased CCL26 and increased IL-6 expression in the nasal and small airway cells, but this effect was not observed in the bronchial cells. IL-6 and CXCL8 were produced in varying degrees by all of the epithelial primary cells in cultures stimulated with TNF-α, LPS or IFN-γ. We showed that epithelial cells from the various levels of the respiratory tract act in a united way, responding in a similar manner to stimulation with IL-4 and IL-13, showing similar reactivity to TNF-α and LPS, and giving an almost unified response to IFN-γ pre-stimulation.

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Regulation of RANTES promoter activation in alveolar epithelial cells after cytokine stimulation

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00162.2002 ◽

2002 ◽

Vol 283 (6) ◽

pp. L1280-L1290 ◽

Cited By ~ 44

Author(s):

Antonella Casola ◽

Allyne Henderson ◽

Tianshuang Liu ◽

Roberto P. Garofalo ◽

Allan R. Brasier

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Viral Infection ◽

Molecular Mechanisms ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Mrna Stabilization ◽

Tnf Α ◽

Ifn Γ ◽

Responsive Element

Regulated on activation, normal T cell expressed, and presumably secreted (RANTES) is a member of the CC chemokine family of proteins implicated in a variety of diseases characterized by lung eosinophilia and inflammation, strongly produced by stimulated airway epithelial cells. Because such cytokines as tumor necrosis factor (TNF)-α and interferon-γ (IFN-γ) have been shown to enhance RANTES induction in airway epithelial cells and RANTES gene expression appears to be differentially regulated depending on the cell type and the stimulus applied, in this study we have elucidated mechanisms that operate to control RANTES induction on exposure to TNF-α and/or IFN-γ. Our results indicate that TNF-α and IFN-γ synergistically induce RANTES protein secretion and mRNA expression. RANTES transcription is activated only after stimulation with TNF-α, but not IFN-γ, which affects RANTES mRNA stabilization. Promoter deletion and mutagenesis experiments indicate that the nuclear factor (NF)-κB site is the most important cis-regulatory element controlling TNF-induced RANTES transcription, although NF-interleukin-6 binding site, cAMP responsive element (CRE), and interferon-stimulated responsive element (ISRE) also play a significant role. TNF-α stimulation induces nuclear translocation of interferon regulatory factor (IRF)-3, which in viral infection binds the RANTES ISRE and is necessary for activation of RANTES transcription. However, TNF-induced IRF-3 translocation does not result in IRF-3 binding to the RANTES ISRE. Although viral infection can activate an ISRE-driven promoter, TNF cannot, indicating that RANTES gene enhancers are controlled in a stimulus-specific fashion. Identification of molecular mechanisms involved in RANTES gene expression is fundamental for developing strategies to modulate lung inflammatory responses.

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TNF, IFN-γ, and endotoxin increase expression of DMT1 in bronchial epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00428.2003 ◽

2005 ◽

Vol 289 (1) ◽

pp. L24-L33 ◽

Cited By ~ 37

Author(s):

Xinchao Wang ◽

Michael D. Garrick ◽

Funmei Yang ◽

Lisa A. Dailey ◽

Claude A. Piantadosi ◽

...

Keyword(s):

Epithelial Cells ◽

Proinflammatory Cytokines ◽

Airway Epithelial Cells ◽

Divalent Metal Transporter 1 ◽

Divalent Metal Transporter ◽

Metal Transporter ◽

Tnf Α ◽

Ifn Γ

Regulation of the metal transport protein divalent metal transporter-1 (DMT1) may contribute to the uptake and detoxification of iron by cells resident in the respiratory tract. Inflammation has been associated with an increased availability of this metal resulting in an oxidative stress. Because proinflammatory cytokines and LPS have been demonstrated to affect an elevated expression of DMT1 in a macrophage cell line, we tested the hypothesis that tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and LPS increase DMT1 expression in airway epithelial cells. We used RT-PCR to detect mRNA for both −IRE DMT1 and +IRE DMT1 in BEAS-2B cells. Treatment with TNF-α, IFN-γ, or LPS increased both forms. Western blot analysis also demonstrated an increase in the expression of both isoforms of DMT1 after these treatments. Twenty-four hours after exposure of an animal model to TNF-α, IFN-γ, or LPS, a significant increase in pulmonary expression of −IRE DMT1 was seen by immunohistochemistry; the level of +IRE DMT1 was too low in the lung to be visualized using this methodology. Finally, iron transport into BEAS-2B cells was increased after inclusion of TNF-α, IFN-γ, or LPS in the media. We conclude that proinflammatory cytokines and LPS increase mRNA and protein expression of DMT1 in airway cells in vitro and in vivo. Furthermore, both −IRE and +IRE isoforms are elevated after exposures. Increased expression of this protein appears to be included in a coordinated response of the cell and tissue where the function might be to diminish availability of metal.

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Effects of TNF‐α, IFN‐γ and IL-β on normal human bronchial epithelial cells

European Respiratory Journal ◽

10.1034/j.1399-3003.1999.14a15.x ◽

1999 ◽

Vol 14 (1) ◽

pp. 84 ◽

Cited By ~ 43

Author(s):

C. Kampf ◽

A.J. Relova ◽

S Sandler ◽

G.M. Roomans

Keyword(s):

Epithelial Cells ◽

Bronchial Epithelial Cells ◽

Normal Human Bronchial Epithelial ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Tnf Α ◽

Normal Human ◽

Ifn Γ

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Chemokine expression in CF epithelia: implications for the role of CFTR in RANTES expression

AJP Cell Physiology ◽

10.1152/ajpcell.1999.276.3.c700 ◽

1999 ◽

Vol 276 (3) ◽

pp. C700-C710 ◽

Cited By ~ 63

Author(s):

Lisa M. Schwiebert ◽

Kim Estell ◽

Stacie M. Propst

Keyword(s):

Epithelial Cells ◽

Nuclear Factor Κb ◽

Interferon Γ ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Monocyte Chemoattractant Protein 1 ◽

Tnf Α ◽

Ifn Γ ◽

Rantes Promoter ◽

Cf Transmembrane Conductance Regulator

To delineate the mechanisms that facilitate leukocyte migration into the cystic fibrosis (CF) lung, expression of chemokines, including interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and RANTES, was compared between CF and non-CF airway epithelia. The findings presented herein demonstrate that, under either basal conditions or tumor necrosis factor-α (TNF-α)- and/or interferon-γ (IFN-γ)-stimulated conditions, a consistent pattern of differences in the secretion of IL-8 and MCP-1 between CF and non-CF epithelial cells was not observed. In contrast, CF epithelial cells expressed no detectable RANTES protein or mRNA under basal conditions or when stimulated with TNF-α and/or IFN-γ ( P ≤ 0.05), unlike their non-CF counterparts. Correction of the CF transmembrane conductance regulator (CFTR) defect in CF airway epithelial cells restored the induction of RANTES protein and mRNA by TNF-α in combination with IFN-γ ( P ≤ 0.05) but had little effect on IL-8 or MCP-1 production compared with mock controls. Transfection studies utilizing RANTES promoter constructs suggested that CFTR activates the RANTES promoter via a nuclear factor-κB-mediated pathway. Together, these results suggest that 1) RANTES expression is altered in CF epithelia and 2) epithelial expression of RANTES, but not IL-8 or MCP-1, is dependent on CFTR.

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Abstract 3699: Multi-walled carbon nanotubes induce apoptosis in normal human small airway epithelial cells through proteasome-mediated Mcl-1 protein degradation

10.1158/1538-7445.am10-3699 ◽

2010 ◽

Author(s):

Bao-Zhu Yuan ◽

Joshua Chapman ◽

Yon Rojanasakul ◽

Vincent Castranova ◽

Steven H. Reynolds

Keyword(s):

Carbon Nanotubes ◽

Epithelial Cells ◽

Protein Degradation ◽

Airway Epithelial Cells ◽

Small Airway ◽

Airway Epithelial ◽

Multi Walled Carbon Nanotubes ◽

Normal Human ◽

Walled Carbon Nanotubes ◽

Small Airway Epithelial Cells

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Ozone effect on inflammatory and proteomic profile of human macrophages and airway epithelial cells

European Journal of Public Health ◽

10.1093/eurpub/ckaa166.102 ◽

2020 ◽

Vol 30 (Supplement_5) ◽

Author(s):

L Falcone ◽

E Aruffo ◽

P Di Carlo ◽

P Del Boccio ◽

M C Cufaro ◽

...

Keyword(s):

Oxidative Stress ◽

Air Pollution ◽

Epithelial Cells ◽

Cell Lines ◽

Urban Areas ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Proteomics Analysis ◽

Human Macrophages ◽

Tnf Α

Abstract Background Reactive oxygen species (ROS) and oxidative stress in the respiratory system are involved in lung inflammation and tumorigenesis. Ozone (O3) is one of the main components of air pollution in urban areas able to act as strong pro-oxidant agent, however its effects on human health is still poorly investigated. In this study the effect of O3 has been evaluated in THP-1 monocytes differentiated into macrophages with PMA and in HBEpC (primary human bronchial epithelial) cells, two model systems for in vitro studies and translational research. Methods Cell viability, ROS and pro-inflammatory cytokines like interleukin-8(IL-8) and tumor necrosis factor(TNF-α) have been tested in the above-mentioned cell lines not exposed to any kind of pollution (basal condition-b.c.) or exposed to O3 at a concentration of 120 ppb. In HBEpC a labelfree shotgun proteomics analysis has been also performed in the same conditions. Results Ozone significantly increased the production of IL-8 and TNF-α in THP-1 whereas no changes were shown in HBEpC. In both cell lines lipopolysaccharide(LPS) caused an increase of IL-8 and TNF-α production in b.c. and O3 treatment potentiated this effect. Ozone exposure increased ROS formation in a time dependent manner in both cell lines and in THP-1 cells a decrease in catalase activity was also shown. Finally, according to these data, functional proteomics analysis revealed that in HBEpC exposure to O3 many differential proteins are related to oxidative stress and inflammation. Conclusions Our results indicate that O3, at levels that can be reached in urban areas, causes an increase of pro-inflammatory agents either per se or potentiating the effect of immune response stimulators in cell models of human macrophages and human airway epithelial cells. Interestingly, the proteomic analysis showed that besides the dysregulated proteins, O3 induced the expression of AKR1D1 and AKR1B10, proteins recognized to play a significant role in cancer development. Key messages This study adds new pieces of information on the association between O3 exposure and detrimental effects on respiratory system. This study suggests the need for further research on the mechanisms involved and for a continued monitoring/re-evaluation of air pollution standards aimed at safeguarding human health.

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Noncoordinated expression of alpha-, beta-, and gamma-subunit mRNAs of epithelial Na+ channel along rat respiratory tract

AJP Cell Physiology ◽

10.1152/ajpcell.1997.272.1.c131 ◽

1997 ◽

Vol 272 (1) ◽

pp. C131-C141 ◽

Cited By ~ 86

Author(s):

N. Farman ◽

C. R. Talbot ◽

R. Boucher ◽

M. Fay ◽

C. Canessa ◽

...

Keyword(s):

Epithelial Cells ◽

Respiratory Tract ◽

Mrna Level ◽

Airway Epithelial Cells ◽

Respiratory Physiology ◽

Na Channel ◽

Alveolar Cells ◽

Cellular Expression ◽

Alpha Beta ◽

Epithelial Na Channel

Na+ reabsorption from the epithelial surface of the respiratory tract plays a fundamental role in respiratory physiology. As in the epithelia of the renal collecting tubule and distal colon, Na+ enters across the luminal surface of respiratory epithelial cells via a recently cloned amiloride-sensitive multisubunit (alpha, beta, gamma) epithelial Na+ channel. We have examined the cellular expression at the mRNA level of the alpha-, beta-, and gamma-subunits of rat epithelial Na+ channel (rENaC) in the rat lung and upper airway epithelial cells using in situ hybridization. A large prevalence of alpha- and gamma-rENaC subunit expression (over beta) was found in tracheal epithelium, in a subpopulation of alveolar cells, presumably type II pneumocytes, and in nasal and tracheal gland acini. In contrast, equivalent levels of expression of all three subunits were detected in bronchiolar epithelium and in rat nasal gland ducts. This diversity of expression may reflect cell-specific functions of the amiloride-sensitive Na+ channel along the respiratory tract.

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