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The multiphasic TNF-α-induced compromise of Calu-3 airway epithelial barrier function
Journal article   Open access   Peer reviewed

The multiphasic TNF-α-induced compromise of Calu-3 airway epithelial barrier function

EXPERIMENTAL LUNG RESEARCH, v 49(1)
31 Dec 2023
PMID: 37000123
url
https://doi.org/10.1080/01902148.2023.2193637View
Published, Version of Record (VoR) Open

Abstract

Purpose: Airway epithelial barrier leak and the involvement of proinflammatory cytokines play a key role in a variety of diseases. This study evaluates barrier compromise by the inflammatory mediator Tumor Necrosis Factor-alpha (TNF-alpha) in the human airway epithelial Calu-3 model. Methods: We examined the effects of TNF-alpha on barrier function in Calu-3 cell layers using Transepithelial Electrical Resistance (TER) and transepithelial diffusion of radiolabeled probe molecules. Western immunoblot analyses of tight junctional (TJ) proteins in detergent soluble fractions were performed. Results: TNF-alpha dramatically reduced TER and increased paracellular permeability of both 14C-D-mannitol and the larger 5 kDa probe, 14C-inulin. A time course of the effects shows two separate actions on barrier function. An initial compromise of barrier function occurs 2-4 hours after TNF-alpha exposure, followed by complete recovery of barrier function by 24 hrs. Beginning 48 hrs. post-exposure, a second more sustained barrier compromise ensues, in which leakiness persists through 144 hrs. There were no changes in TJ proteins observed at 3 hrs. post exposure, but significant increases in claudins-2, -3, -4, and -5, as well as a decrease in occludin were seen at 72 hrs. post TNF-alpha exposure. Both the 2-4 hr. and the 72 hr. TNF-alpha induced leaks are shown to be mediated by the ERK signaling pathway. Conclusion: TNF-alpha induced a multiphasic transepithelial leak in Calu-3 cell layers that was shown to be ERK mediated, as well as involve changes in the TJ complex. The micronutrients, retinoic acid and calcitriol, were effective at reducing this barrier compromise caused by TNF-alpha. The significance of these results for airway disease and for COVID-19 specifically are discussed.

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Collaboration types
Domestic collaboration
Web of Science research areas
Respiratory System
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