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Cellular crosstalk between airway epithelial and endothelial cells regulates barrier functions during exposure to double-stranded RNA

Cellular crosstalk between airway epithelial and endothelial cells regulates barrier functions during exposure to double-stranded RNA
Cellular crosstalk between airway epithelial and endothelial cells regulates barrier functions during exposure to double-stranded RNA
Introduction: The epithelial and endothelial barriers of the airway mucosa are critical for regulation of tissue homeostasis and protection against pathogens or other tissue damaging agents. In response to a viral infection, epithelial cells must signal to the endothelium to initiate immune cell recruitment. This is a highly temporal regulated process; however, the mechanisms of this cross-talk are not fully understood.

Methods: In a close-contact co-culture model of human airway epithelial and endothelial cells cellular crosstalk was analysed using transepithelial electrical resistance (TER) measurements, immunofluorescence, electron microscopy and ELISA. Viral infections were simulated by exposing airway epithelial cells apically to double-stranded RNA (Poly(I:C)). Using a microfluidic culture system the temporal release of mediators was analysed in the co-culture model.

Results: Within 4h of challenge, double-stranded RNA induced the release of TNF-a by epithelial cells. This activated endothelial cells by triggering the release of the chemoattractant CX3CL1 (fractalkine) by 8h post-challenge and expression of adhesion molecules E-selectin and ICAM-1. These responses were significantly reduced by neutralising TNF-a.

Conclusion: By facilitating kinetic profiling, the microfluidic co-culture system has enabled identification of a key signalling mechanism between the epithelial and endothelial barriers. Better understanding of cell-cell cross-talk and its regulatory mechanisms has the potential to identify new therapeutic strategies to control airway inflammation.
2050-4527
45-56
Blume, Cornelia
aa391c64-8718-4238-906b-d6bb1551a07b
Reale, Riccardo
c7651c37-e622-45aa-a0e1-595d35ca4b2c
Held, Marie
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Loxham, Matthew
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Millar, Tim
ec88510c-ad88-49f6-8b2d-4277c84c1958
Collins, Jane
be0e66f1-3036-47fa-9d7e-914c48710ba4
Swindle, Emily
fe393c7a-a513-4de4-b02e-27369bd7e84f
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Davies, Donna
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Blume, Cornelia
aa391c64-8718-4238-906b-d6bb1551a07b
Reale, Riccardo
c7651c37-e622-45aa-a0e1-595d35ca4b2c
Held, Marie
b883239a-b16c-401a-a6be-3705faaef229
Loxham, Matthew
8ef02171-9040-4c1d-8452-2ca34c56facb
Millar, Tim
ec88510c-ad88-49f6-8b2d-4277c84c1958
Collins, Jane
be0e66f1-3036-47fa-9d7e-914c48710ba4
Swindle, Emily
fe393c7a-a513-4de4-b02e-27369bd7e84f
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Davies, Donna
7de8fdc7-3640-4e3a-aa91-d0e03f990c38

Blume, Cornelia, Reale, Riccardo, Held, Marie, Loxham, Matthew, Millar, Tim, Collins, Jane, Swindle, Emily, Morgan, Hywel and Davies, Donna (2017) Cellular crosstalk between airway epithelial and endothelial cells regulates barrier functions during exposure to double-stranded RNA. Immunity, Inflammation and Disease, 5 (1), 45-56. (doi:10.1002/iid3.139).

Record type: Article

Abstract

Introduction: The epithelial and endothelial barriers of the airway mucosa are critical for regulation of tissue homeostasis and protection against pathogens or other tissue damaging agents. In response to a viral infection, epithelial cells must signal to the endothelium to initiate immune cell recruitment. This is a highly temporal regulated process; however, the mechanisms of this cross-talk are not fully understood.

Methods: In a close-contact co-culture model of human airway epithelial and endothelial cells cellular crosstalk was analysed using transepithelial electrical resistance (TER) measurements, immunofluorescence, electron microscopy and ELISA. Viral infections were simulated by exposing airway epithelial cells apically to double-stranded RNA (Poly(I:C)). Using a microfluidic culture system the temporal release of mediators was analysed in the co-culture model.

Results: Within 4h of challenge, double-stranded RNA induced the release of TNF-a by epithelial cells. This activated endothelial cells by triggering the release of the chemoattractant CX3CL1 (fractalkine) by 8h post-challenge and expression of adhesion molecules E-selectin and ICAM-1. These responses were significantly reduced by neutralising TNF-a.

Conclusion: By facilitating kinetic profiling, the microfluidic co-culture system has enabled identification of a key signalling mechanism between the epithelial and endothelial barriers. Better understanding of cell-cell cross-talk and its regulatory mechanisms has the potential to identify new therapeutic strategies to control airway inflammation.

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Blume et al, 2016 Immunity, Inflammation and Disease accepted manuscript - Accepted Manuscript
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Blume et al 2017 Immunity, Inflammation and Disease - Version of Record
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Accepted/In Press date: 26 October 2016
e-pub ahead of print date: 18 January 2017
Published date: March 2017
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 402168
URI: https://eprints.soton.ac.uk/id/eprint/402168
ISSN: 2050-4527
PURE UUID: 4558c324-271d-4786-b4fc-50774ef13050
ORCID for Cornelia Blume: ORCID iD orcid.org/0000-0001-6133-7318
ORCID for Matthew Loxham: ORCID iD orcid.org/0000-0001-6459-538X
ORCID for Tim Millar: ORCID iD orcid.org/0000-0002-4539-2445
ORCID for Emily Swindle: ORCID iD orcid.org/0000-0003-3644-7747
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676
ORCID for Donna Davies: ORCID iD orcid.org/0000-0002-5117-2991

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Date deposited: 31 Oct 2016 16:53
Last modified: 10 Dec 2019 06:21

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Contributors

Author: Cornelia Blume ORCID iD
Author: Riccardo Reale
Author: Marie Held
Author: Matthew Loxham ORCID iD
Author: Tim Millar ORCID iD
Author: Jane Collins
Author: Emily Swindle ORCID iD
Author: Hywel Morgan ORCID iD
Author: Donna Davies ORCID iD

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