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Pro-inflammatory actions of the exodomain shed from Protease Activated Receptor 2 (PAR-2)

Pro-inflammatory actions of the exodomain shed from Protease Activated Receptor 2 (PAR-2)
Pro-inflammatory actions of the exodomain shed from Protease Activated Receptor 2 (PAR-2)
Rationale:
Activation of PAR-2 by proteases has been implicated as a key mechanism in allergic and other inflammatory conditions. Proteolytic cleavage can lead to exposure of a tethered ligand whose binding to the receptor stimulates cell signalling, and the release from PAR-2 of an exodomain fragment (PAR-2E). Potential functions of PAR-2E have been little studied.

Methods:
PAR-2E was synthesised and its stability investigated in human biological fluids, cell supernatants and with selected proteases. The ability of PAR-2E to stimulate calcium flux in cultures of human umbilical vein endothelial cells was examined using a fluorescence based microplate procedure, and altered RNA expression by whole genome microarray analysis and quantitative PCR (qPCR) for selected genes. Adhesion molecule expression was investigated by flow cytometry. In parallel studies, nucleated cells were enumerated and levels of matrix metalloproteases (MMP) determined by gelatin zymography in peritoneal lavage fluid from C57BL/6 mice injected intraperitoneally with PAR-2E.

Results:
PAR-2E was highly susceptible to degradation by proteases, but its addition to cells resulted in calcium flux, and increased expression of genes for various adhesion molecules (including ICAM1, EPCAM and ITGAL), and cytokines (TNFAIP3 and IL1B). PAR-2E-induced upregulation of ICAM1, VCAM-1, EPCAM and ITGAL was observed on flow cytometry. Injection of PAR-2E into mice was associated with eosinophilia and raised levels of MMP2 in peritoneal lavage fluid.

Conclusions:
PAR-2E may act as a stimulus for increased expression of adhesion molecules, cytokine release and eosinophilia, and deserves consideration as a mediator of inflammation following PAR-2 activation.
0091-6749
AB47
Khedr, Mogib E.
37c876ff-7226-4d51-bd20-dfe0bbca3d73
Abdelmotelb, Ahmed
cebf8ef6-9dbe-433d-b8ac-f9f8c0ee3f94
Lau, Laurie
2af8045d-6162-4939-aba7-28dd2f60f6a8
Arno, Matthew
77edce96-78e5-49c9-92b0-f91a69eefb9b
Pender, Sylvia
62528b03-ec42-41bb-80fe-48454c2c5242
Zhou, Xiaoying
84558a96-3129-44de-b295-869d9ee4d19f
Walls, Andrew F.
aaa7e455-0562-4b4c-94f5-ec29c74b1bfe
Khedr, Mogib E.
37c876ff-7226-4d51-bd20-dfe0bbca3d73
Abdelmotelb, Ahmed
cebf8ef6-9dbe-433d-b8ac-f9f8c0ee3f94
Lau, Laurie
2af8045d-6162-4939-aba7-28dd2f60f6a8
Arno, Matthew
77edce96-78e5-49c9-92b0-f91a69eefb9b
Pender, Sylvia
62528b03-ec42-41bb-80fe-48454c2c5242
Zhou, Xiaoying
84558a96-3129-44de-b295-869d9ee4d19f
Walls, Andrew F.
aaa7e455-0562-4b4c-94f5-ec29c74b1bfe

Khedr, Mogib E., Abdelmotelb, Ahmed, Lau, Laurie, Arno, Matthew, Pender, Sylvia, Zhou, Xiaoying and Walls, Andrew F. (2013) Pro-inflammatory actions of the exodomain shed from Protease Activated Receptor 2 (PAR-2). Journal of Allergy and Clinical Immunology, 131 (2, Supplment), AB47. (doi:10.1016/j.jaci.2012.12.851).

Record type: Meeting abstract

Abstract

Rationale:
Activation of PAR-2 by proteases has been implicated as a key mechanism in allergic and other inflammatory conditions. Proteolytic cleavage can lead to exposure of a tethered ligand whose binding to the receptor stimulates cell signalling, and the release from PAR-2 of an exodomain fragment (PAR-2E). Potential functions of PAR-2E have been little studied.

Methods:
PAR-2E was synthesised and its stability investigated in human biological fluids, cell supernatants and with selected proteases. The ability of PAR-2E to stimulate calcium flux in cultures of human umbilical vein endothelial cells was examined using a fluorescence based microplate procedure, and altered RNA expression by whole genome microarray analysis and quantitative PCR (qPCR) for selected genes. Adhesion molecule expression was investigated by flow cytometry. In parallel studies, nucleated cells were enumerated and levels of matrix metalloproteases (MMP) determined by gelatin zymography in peritoneal lavage fluid from C57BL/6 mice injected intraperitoneally with PAR-2E.

Results:
PAR-2E was highly susceptible to degradation by proteases, but its addition to cells resulted in calcium flux, and increased expression of genes for various adhesion molecules (including ICAM1, EPCAM and ITGAL), and cytokines (TNFAIP3 and IL1B). PAR-2E-induced upregulation of ICAM1, VCAM-1, EPCAM and ITGAL was observed on flow cytometry. Injection of PAR-2E into mice was associated with eosinophilia and raised levels of MMP2 in peritoneal lavage fluid.

Conclusions:
PAR-2E may act as a stimulus for increased expression of adhesion molecules, cytokine release and eosinophilia, and deserves consideration as a mediator of inflammation following PAR-2 activation.

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More information

Published date: February 2013
Venue - Dates: American Academy of Allergy, Asthma & Immunology (AAAAI): 2013 AAAAI Annual Meeting, , San Antonio, United States, 2013-02-22 - 2013-02-26

Identifiers

Local EPrints ID: 432850
URI: http://eprints.soton.ac.uk/id/eprint/432850
ISSN: 0091-6749
PURE UUID: cf70b102-ead3-4d69-bad9-a0ebe80e182d
ORCID for Mogib E. Khedr: ORCID iD orcid.org/0000-0001-9942-4409
ORCID for Sylvia Pender: ORCID iD orcid.org/0000-0001-6332-0333
ORCID for Andrew F. Walls: ORCID iD orcid.org/0000-0003-4803-4595

Catalogue record

Date deposited: 31 Jul 2019 16:30
Last modified: 10 Nov 2021 03:32

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Contributors

Author: Mogib E. Khedr ORCID iD
Author: Ahmed Abdelmotelb
Author: Laurie Lau
Author: Matthew Arno
Author: Sylvia Pender ORCID iD
Author: Xiaoying Zhou
Author: Andrew F. Walls ORCID iD

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