A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung

Arlene M A Glasgow; Donna M Small; Aaron Scott; Denise T McLean; Nicolas Camper; Umar Hamid; Shauna Hegarty; Dhruv Parekh; Cecilia O'Kane; Fionnuala T Lundy; Paul McNally; J Stuart Elborn; Danny F McAuley; Sinéad Weldon; Clifford C Taggart

doi:10.1136/thoraxjnl-2014-206488

A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung

Arlene M A Glasgow, Donna M Small, Aaron Scott, Denise T McLean, Nicolas Camper, Umar Hamid, Shauna Hegarty, Dhruv Parekh, Cecilia O'Kane, Fionnuala T Lundy, Paul McNally, J Stuart Elborn, Danny F McAuley, Sinéad Weldon, Clifford C Taggart

Inflammation and Ageing

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

Abstract

INTRODUCTION: Secretory leucocyte protease inhibitor and elafin are members of the whey acidic protein (WAP), or WAP four disulfide-core (WFDC), family of proteins and have multiple contributions to innate defence including inhibition of neutrophil serine proteases and inhibition of the inflammatory response to lipopolysaccharide (LPS). This study aimed to explore potential activities of WFDC12, a previously uncharacterised WFDC protein expressed in the lung.

METHODS: Recombinant expression and purification of WFDC12 were optimised in Escherichia coli. Antiprotease, antibacterial and immunomodulatory activities of recombinant WFDC12 were evaluated and levels of endogenous WFDC12 protein were characterised by immunostaining and ELISA.

RESULTS: Recombinant WFDC12 inhibited cathepsin G, but not elastase or proteinase-3 activity. Monocytic cells pretreated with recombinant WFDC12 before LPS stimulation produced significantly lower levels of the pro-inflammatory cytokines interleukin-8 and monocyte chemotactic protein-1 compared with cells stimulated with LPS alone. Recombinant WFDC12 became conjugated to fibronectin in a transglutaminase-mediated reaction and retained antiprotease activity. In vivo WFDC12 expression was confirmed by immunostaining of human lung tissue sections. WFDC12 levels in human bronchoalveolar lavage fluid from healthy and lung-injured patients were quantitatively compared, showing WFDC12 to be elevated in both patients with acute respiratory distress syndrome and healthy subjects treated with LPS, relative to healthy controls.

CONCLUSIONS: Together, these results suggest a role for this lesser known WFDC protein in the regulation of lung inflammation.

Original language	English
Pages (from-to)	426-432
Number of pages	7
Journal	Thorax
Volume	70
Issue number	5
Early online date	13 Mar 2015
DOIs	https://doi.org/10.1136/thoraxjnl-2014-206488
Publication status	Published - May 2015

Keywords

Bronchoalveolar Lavage Fluid
Humans
Lipopolysaccharides
Lung
Microbial Sensitivity Tests
Monocytes
Proteins
Recombinant Proteins
Serine Endopeptidases
Tissue Culture Techniques

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Glasgow, A. M. A., Small, D. M., Scott, A., McLean, D. T., Camper, N., Hamid, U., Hegarty, S., Parekh, D., O'Kane, C., Lundy, F. T., McNally, P., Elborn, J. S., McAuley, D. F., Weldon, S., & Taggart, C. C. (2015). A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung. Thorax, 70(5), 426-432. https://doi.org/10.1136/thoraxjnl-2014-206488

@article{f63bdecab1b5463face544335c37655b,

title = "A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung",

abstract = "INTRODUCTION: Secretory leucocyte protease inhibitor and elafin are members of the whey acidic protein (WAP), or WAP four disulfide-core (WFDC), family of proteins and have multiple contributions to innate defence including inhibition of neutrophil serine proteases and inhibition of the inflammatory response to lipopolysaccharide (LPS). This study aimed to explore potential activities of WFDC12, a previously uncharacterised WFDC protein expressed in the lung.METHODS: Recombinant expression and purification of WFDC12 were optimised in Escherichia coli. Antiprotease, antibacterial and immunomodulatory activities of recombinant WFDC12 were evaluated and levels of endogenous WFDC12 protein were characterised by immunostaining and ELISA.RESULTS: Recombinant WFDC12 inhibited cathepsin G, but not elastase or proteinase-3 activity. Monocytic cells pretreated with recombinant WFDC12 before LPS stimulation produced significantly lower levels of the pro-inflammatory cytokines interleukin-8 and monocyte chemotactic protein-1 compared with cells stimulated with LPS alone. Recombinant WFDC12 became conjugated to fibronectin in a transglutaminase-mediated reaction and retained antiprotease activity. In vivo WFDC12 expression was confirmed by immunostaining of human lung tissue sections. WFDC12 levels in human bronchoalveolar lavage fluid from healthy and lung-injured patients were quantitatively compared, showing WFDC12 to be elevated in both patients with acute respiratory distress syndrome and healthy subjects treated with LPS, relative to healthy controls.CONCLUSIONS: Together, these results suggest a role for this lesser known WFDC protein in the regulation of lung inflammation.",

keywords = "Bronchoalveolar Lavage Fluid, Humans, Lipopolysaccharides, Lung, Microbial Sensitivity Tests, Monocytes, Proteins, Recombinant Proteins, Serine Endopeptidases, Tissue Culture Techniques",

author = "Glasgow, {Arlene M A} and Small, {Donna M} and Aaron Scott and McLean, {Denise T} and Nicolas Camper and Umar Hamid and Shauna Hegarty and Dhruv Parekh and Cecilia O'Kane and Lundy, {Fionnuala T} and Paul McNally and Elborn, {J Stuart} and McAuley, {Danny F} and Sin{\'e}ad Weldon and Taggart, {Clifford C}",

note = "Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.",

year = "2015",

month = may,

doi = "10.1136/thoraxjnl-2014-206488",

language = "English",

volume = "70",

pages = "426--432",

journal = "Thorax",

issn = "0040-6376",

publisher = "BMJ Publishing Group",

number = "5",

}

Glasgow, AMA, Small, DM, Scott, A, McLean, DT, Camper, N, Hamid, U, Hegarty, S, Parekh, D, O'Kane, C, Lundy, FT, McNally, P, Elborn, JS, McAuley, DF, Weldon, S & Taggart, CC 2015, 'A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung', Thorax, vol. 70, no. 5, pp. 426-432. https://doi.org/10.1136/thoraxjnl-2014-206488

TY - JOUR

T1 - A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung

AU - Glasgow, Arlene M A

AU - Small, Donna M

AU - Scott, Aaron

AU - McLean, Denise T

AU - Camper, Nicolas

AU - Hamid, Umar

AU - Hegarty, Shauna

AU - Parekh, Dhruv

AU - O'Kane, Cecilia

AU - Lundy, Fionnuala T

AU - McNally, Paul

AU - Elborn, J Stuart

AU - McAuley, Danny F

AU - Weldon, Sinéad

AU - Taggart, Clifford C

N1 - Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

PY - 2015/5

Y1 - 2015/5

N2 - INTRODUCTION: Secretory leucocyte protease inhibitor and elafin are members of the whey acidic protein (WAP), or WAP four disulfide-core (WFDC), family of proteins and have multiple contributions to innate defence including inhibition of neutrophil serine proteases and inhibition of the inflammatory response to lipopolysaccharide (LPS). This study aimed to explore potential activities of WFDC12, a previously uncharacterised WFDC protein expressed in the lung.METHODS: Recombinant expression and purification of WFDC12 were optimised in Escherichia coli. Antiprotease, antibacterial and immunomodulatory activities of recombinant WFDC12 were evaluated and levels of endogenous WFDC12 protein were characterised by immunostaining and ELISA.RESULTS: Recombinant WFDC12 inhibited cathepsin G, but not elastase or proteinase-3 activity. Monocytic cells pretreated with recombinant WFDC12 before LPS stimulation produced significantly lower levels of the pro-inflammatory cytokines interleukin-8 and monocyte chemotactic protein-1 compared with cells stimulated with LPS alone. Recombinant WFDC12 became conjugated to fibronectin in a transglutaminase-mediated reaction and retained antiprotease activity. In vivo WFDC12 expression was confirmed by immunostaining of human lung tissue sections. WFDC12 levels in human bronchoalveolar lavage fluid from healthy and lung-injured patients were quantitatively compared, showing WFDC12 to be elevated in both patients with acute respiratory distress syndrome and healthy subjects treated with LPS, relative to healthy controls.CONCLUSIONS: Together, these results suggest a role for this lesser known WFDC protein in the regulation of lung inflammation.

AB - INTRODUCTION: Secretory leucocyte protease inhibitor and elafin are members of the whey acidic protein (WAP), or WAP four disulfide-core (WFDC), family of proteins and have multiple contributions to innate defence including inhibition of neutrophil serine proteases and inhibition of the inflammatory response to lipopolysaccharide (LPS). This study aimed to explore potential activities of WFDC12, a previously uncharacterised WFDC protein expressed in the lung.METHODS: Recombinant expression and purification of WFDC12 were optimised in Escherichia coli. Antiprotease, antibacterial and immunomodulatory activities of recombinant WFDC12 were evaluated and levels of endogenous WFDC12 protein were characterised by immunostaining and ELISA.RESULTS: Recombinant WFDC12 inhibited cathepsin G, but not elastase or proteinase-3 activity. Monocytic cells pretreated with recombinant WFDC12 before LPS stimulation produced significantly lower levels of the pro-inflammatory cytokines interleukin-8 and monocyte chemotactic protein-1 compared with cells stimulated with LPS alone. Recombinant WFDC12 became conjugated to fibronectin in a transglutaminase-mediated reaction and retained antiprotease activity. In vivo WFDC12 expression was confirmed by immunostaining of human lung tissue sections. WFDC12 levels in human bronchoalveolar lavage fluid from healthy and lung-injured patients were quantitatively compared, showing WFDC12 to be elevated in both patients with acute respiratory distress syndrome and healthy subjects treated with LPS, relative to healthy controls.CONCLUSIONS: Together, these results suggest a role for this lesser known WFDC protein in the regulation of lung inflammation.

KW - Bronchoalveolar Lavage Fluid

KW - Humans

KW - Lipopolysaccharides

KW - Lung

KW - Microbial Sensitivity Tests

KW - Monocytes

KW - Proteins

KW - Recombinant Proteins

KW - Serine Endopeptidases

KW - Tissue Culture Techniques

U2 - 10.1136/thoraxjnl-2014-206488

DO - 10.1136/thoraxjnl-2014-206488

M3 - Article

C2 - 25770093

SN - 0040-6376

VL - 70

SP - 426

EP - 432

JO - Thorax

JF - Thorax

IS - 5

ER -

A role for whey acidic protein four-disulfide-core 12 (WFDC12) in the regulation of the inflammatory response in the lung

Abstract

Keywords

UN SDGs

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