Diesel exhaust particles modulate the tight junction protein occludin in lung cells in vitro

Abstract Background: While exposure to diesel exhaust particles has been linked to aberrant immune responses in allergic diseases such as asthma, little attention has been paid to their effects on the airway epithelium. In this study, we sought to determine the effect of diesel exhaust exposure on airway epithelial barrier function and composition using in vitro and in vivo model systems. Methods: 16HBE14o- human bronchial epithelial cells were grown on collagen coated Transwell inserts and exposed to 5 to 50 µg/cm2 SRM 2975 diesel particulate matter (DEP) suspended in cell culture medium or vehicle controls. Changes in barrier function were assessed by measuring transepithelial electrical resistance (TEER) and permeability to 4 kDa FITC Dextran. Neonatal BALB/c mice were exposed to aerosolized DEP (255 ± 89 µg/m3; 2 hours per day for 5 days) and changes in the tight junction protein Tricellulin were assessed two weeks post exposure. Results: A six-hour incubation of epithelial cells with diesel exhaust particles caused a significant concentration-dependent reduction in epithelial barrier integrity as measured by decreased TEER and increased permeability to 4 kDa FITC-Dextran. This reduction in epithelial barrier integrity corresponded to a significant reduction in expression of the tight junction protein Tricellulin. siRNA mediated knockdown of Tricellulin recapitulated changes in barrier function caused by DEP exposure. Neonatal exposure to aerosolized DEP caused a significant reduction in lung Tricellulin two weeks post exposure at both the protein and mRNA level. Conclusion: Short term exposure to DEP causes a significant reduction in epithelial barrier integrity through a reduction in the tight junction protein Tricellulin. Neonatal exposure to aerosolized DEP caused a significant and sustained reduction in Tricellulin protein and mRNA in the lung, suggesting that early life exposure to inhaled DEP may cause lasting changes in airway epithelial barrier function.

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Diesel Exhaust Particle Exposure Reduces Expression of the Epithelial Tight Junction Protein Tricellulin

10.21203/rs.3.rs-25522/v1 ◽

2020 ◽

Author(s):

Timothy Smyth ◽

Janelle Veazey ◽

Sophia Eliseeva ◽

David Chalupa ◽

Alison Elder ◽

...

Keyword(s):

Tight Junction ◽

Diesel Exhaust ◽

Barrier Function ◽

Diesel Exhaust Particles ◽

Tight Junction Protein ◽

Epithelial Barrier ◽

Airway Epithelial ◽

Epithelial Barrier Function ◽

Barrier Integrity ◽

Junction Protein

Abstract Background While exposure to diesel exhaust particles has been linked to aberrant immune responses in allergic diseases such as asthma, little attention has been payed to their effects on the airway epithelium. In this study, we sought to determine the effect of diesel exhaust exposure on airway epithelial barrier function and composition using in vitro and in vivo model systems. 16HBE14o- human bronchial epithelial cells were grown on collagen coated Transwell inserts and exposed to 5 to 50 µg/cm2 SRM 2975 diesel particulate matter (DEP) suspended in cell culture medium or vehicle controls. Changes in barrier function were assessed by measuring transepithelial electrical resistance (TEER) and permeability to 4 kDa FITC Dextran. Neonatal BALB/c mice were exposed to aerosolized DEP (255 ± 89 µg/m3; 2 hours per day for 5 days) and changes in the tight junction protein Tricellulin were assessed two weeks post exposure. Results A six-hour incubation of epithelial cells with diesel exhaust particles caused a significant concentration-dependent reduction in epithelial barrier integrity as measured by decreased TEER and increased permeability to 4 kDa FITC-Dextran. This reduction in epithelial barrier integrity corresponded to a significant reduction in expression of the tight junction protein Tricellulin. siRNA mediated knockdown of Tricellulin recapitulated changes in barrier function caused by DEP exposure. Neonatal exposure to aerosolized DEP caused a significant reduction in lung Tricellulin two weeks post exposure at both the protein and mRNA level. Conclusion Short term exposure to DEP causes a significant reduction in epithelial barrier integrity through a reduction in the tight junction protein Tricellulin. Neonatal exposure to aerosolized DEP caused a significant and sustained reduction in Tricellulin protein and mRNA in the lung, suggesting that early life exposure to inhaled DEP may cause lasting changes in airway epithelial barrier function.

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Diesel exhaust particle exposure reduces expression of the epithelial tight junction protein Tricellulin

Particle and Fibre Toxicology ◽

10.1186/s12989-020-00383-x ◽

2020 ◽

Vol 17 (1) ◽

Author(s):

Timothy Smyth ◽

Janelle Veazey ◽

Sophia Eliseeva ◽

David Chalupa ◽

Alison Elder ◽

...

Keyword(s):

Tight Junction ◽

Diesel Exhaust ◽

Barrier Function ◽

Diesel Exhaust Particles ◽

Tight Junction Protein ◽

Epithelial Barrier ◽

Airway Epithelial ◽

Epithelial Barrier Function ◽

Barrier Integrity ◽

Junction Protein

Abstract Background While exposure to diesel exhaust particles has been linked to aberrant immune responses in allergic diseases such as asthma, little attention has been paid to their effects on the airway epithelial barrier. In this study, we sought to determine the effect of diesel exhaust exposure on airway epithelial barrier function and composition using in vitro and in vivo model systems. Methods 16HBE14o- human bronchial epithelial cells were grown on collagen coated Transwell inserts and exposed to 5 to 50 μg/cm2 SRM 2975 diesel particulate matter (DEP) suspended in cell culture medium or vehicle controls. Changes in barrier function were assessed by measuring transepithelial electrical resistance (TEER) and permeability to 4 kDa FITC Dextran. Neonatal BALB/c mice were exposed to aerosolized DEP (255 ± 89 μg/m3; 2 h per day for 5 days) and changes in the tight junction protein Tricellulin were assessed 2 weeks post exposure. Results A six-hour incubation of epithelial cells with diesel exhaust particles caused a significant concentration-dependent reduction in epithelial barrier integrity as measured by decreased TEER and increased permeability to 4 kDa FITC-Dextran. This reduction in epithelial barrier integrity corresponded to a significant reduction in expression of the tight junction protein Tricellulin. siRNA mediated knockdown of Tricellulin recapitulated changes in barrier function caused by DEP exposure. Neonatal exposure to aerosolized DEP caused a significant reduction in lung Tricellulin 2 weeks post exposure at both the protein and mRNA level. Conclusion Short term exposure to DEP causes a significant reduction in epithelial barrier integrity through a reduction in the tight junction protein Tricellulin. Neonatal exposure to aerosolized DEP caused a significant and sustained reduction in Tricellulin protein and mRNA in the lung, suggesting that early life exposure to inhaled DEP may cause lasting changes in airway epithelial barrier function.

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Diesel Exhaust Particle Exposure Reduces Expression of the Epithelial Tight Junction Protein Tricellulin

10.21203/rs.3.rs-25522/v3 ◽

2020 ◽

Author(s):

Timothy Smyth ◽

Janelle Veazey ◽

Sophia Eliseeva ◽

David Chalupa ◽

Alison Elder ◽

...

Keyword(s):

Tight Junction ◽

Diesel Exhaust ◽

Barrier Function ◽

Diesel Exhaust Particles ◽

Tight Junction Protein ◽

Epithelial Barrier ◽

Airway Epithelial ◽

Epithelial Barrier Function ◽

Barrier Integrity ◽

Junction Protein

Abstract Background: While exposure to diesel exhaust particles has been linked to aberrant immune responses in allergic diseases such as asthma, little attention has been paid to their effects on the airway epithelial barrier. In this study, we sought to determine the effect of diesel exhaust exposure on airway epithelial barrier function and composition using in vitro and in vivo model systems. Methods: 16HBE14o- human bronchial epithelial cells were grown on collagen coated Transwell inserts and exposed to 5 to 50 µg/cm 2 SRM 2975 diesel particulate matter (DEP) suspended in cell culture medium or vehicle controls. Changes in barrier function were assessed by measuring transepithelial electrical resistance (TEER) and permeability to 4 kDa FITC Dextran. Neonatal BALB/c mice were exposed to aerosolized DEP (255 ±89 µg/m 3 ; 2 hours per day for 5 days) and changes in the tight junction protein Tricellulin were assessed two weeks post exposure. Results: A six-hour incubation of epithelial cells with diesel exhaust particles caused a significant concentration-dependent reduction in epithelial barrier integrity as measured by decreased TEER and increased permeability to 4 kDa FITC-Dextran. This reduction in epithelial barrier integrity corresponded to a significant reduction in expression of the tight junction protein Tricellulin. siRNA mediated knockdown of Tricellulin recapitulated changes in barrier function caused by DEP exposure. Neonatal exposure to aerosolized DEP caused a significant reduction in lung Tricellulin two weeks post exposure at both the protein and mRNA level. Conclusion: Short term exposure to DEP causes a significant reduction in epithelial barrier integrity through a reduction in the tight junction protein Tricellulin. Neonatal exposure to aerosolized DEP caused a significant and sustained reduction in Tricellulin protein and mRNA in the lung, suggesting that early life exposure to inhaled DEP may cause lasting changes in airway epithelial barrier function.

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Diesel exhaust particles induced release of interleukin 6 and 8 by (primed) human bronchial epithelial cells (BEAS 2b) in vitro

Immunology Letters ◽

10.1016/s0165-2478(97)87582-7 ◽

1997 ◽

Vol 56 (1-3) ◽

pp. 185

Author(s):

P Steerenberg

Keyword(s):

Epithelial Cells ◽

Diesel Exhaust ◽

Interleukin 6 ◽

Bronchial Epithelial Cells ◽

Diesel Exhaust Particles ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Exhaust Particles

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Comparison of ciliary activity and inflammatory mediator release from bronchial epithelial cells of nonatopic nonasthmatic subjects and atopic asthmatic patients and the effect of diesel exhaust particles in vitro☆☆☆★★★

Journal of Allergy and Clinical Immunology ◽

10.1016/s0091-6749(98)70017-x ◽

1998 ◽

Vol 102 (5) ◽

pp. 771-782 ◽

Cited By ~ 63

Author(s):

Hasan Bayram ◽

Jagdish L. Devalia ◽

Omar A. Khair ◽

Muntasir M. Abdelaziz ◽

Raymond J. Sapsford ◽

...

Keyword(s):

Epithelial Cells ◽

Diesel Exhaust ◽

Inflammatory Mediator ◽

Diesel Exhaust Particles ◽

Mediator Release ◽

Ciliary Activity ◽

Bronchial Epithelial ◽

Asthmatic Patients ◽

Exhaust Particles

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Gene Expression Analysis to Investigate Biological Networks Underlying Nasal Inflammatory Dysfunctions Induced by Diesel Exhaust Particles Using an In Vivo System

Annals of Otology Rhinology & Laryngology ◽

10.1177/0003489419883289 ◽

2019 ◽

Vol 129 (3) ◽

pp. 245-255 ◽

Cited By ~ 1

Author(s):

Hyun Soo Kim ◽

Byeong-Gon Kim ◽

Sohyeon Park ◽

Nahyun Kim ◽

An-Soo Jang ◽

...

Keyword(s):

Gene Expression ◽

Signaling Pathways ◽

Diesel Exhaust ◽

Expression Profiles ◽

Complex Mixture ◽

Diesel Exhaust Particles ◽

The Body ◽

Exhaust Particles

Objectives: Diesel exhaust particles (DEP)s are notorious ambient pollutants composed of a complex mixture of a carbon core and diverse chemical irritants. Several studies have demonstrated significant relationships between DEP exposure and serious nasal inflammatory response in vitro, but available information regarding underlying networks in terms of gene expression changes has not sufficiently explained potential mechanisms of DEP-induced nasal damage, especially in vivo. Methods: In the present study, we identified DEP-induced gene expression profiles under short-term and long-term exposure, and identified signaling pathways based on microarray data for understanding effects of DEP exposure in the mouse nasal cavity. Results: Alteration in gene expression due to DEP exposure provokes an imbalance of the immune system via dysregulated inflammatory markers, predicted to disrupt protective responses against harmful exogenous substances in the body. Several candidate markers were identified after validation using qRT-PCR, including S100A9, CAMP, IL20, and S100A8. Conclusions: Although further mechanistic studies are required for verifying the utility of the potential biomarkers suggested by the present study, our in vivo results may provide meaningful suggestions for understanding the complex cellular signaling pathways involved in DEP-induced nasal damages.

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