RT Journal Article
SR Electronic
T1 A role for the WFDC protein, WAP2, in regulation of innate host defence in the lung
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP 4850
VO 44
IS Suppl 58
A1 Arlene Glasgow
A1 Donna Small
A1 Aaron Scott
A1 Denise McLean
A1 Nicolas Camper
A1 Umar Hamid
A1 Shauna Hegarty
A1 Cecilia O'Kane
A1 Danny McAuley
A1 Fionnuala Lundy
A1 Stuart Elborn
A1 Sinéad Weldon
A1 Cliff Taggart
YR 2014
UL http://erj.ersjournals.com/content/44/Suppl_58/4850.abstract
AB Secretory leukocyte protease inhibitor (SLPI) and elafin are members of the WAP Four-Disulphide-Core (WFDC) family of proteins and have multiple contributions to innate immunity including inhibition of neutrophil serine proteases, antimicrobial properties and inhibition of the inflammatory response to LPS. The aims of this research were to explore potential activities of WAP2, a previously uncharacterised WFDC protein expressed in the lung. Recombinant expression and purification of WAP2 were optimised in E. coli for use in a range of functional assays. Protease activity assays were used to test antiprotease activity of rWAP2. THP-1 monocytic cells were stimulated with LPS alone or rWAP2 in combination with LPS; cytokine levels in cell-free supernatants were subsequently analysed by ELISA. To test if WAP2 could become cross-linked to extracellular matrix proteins, rWAP2 was incubated with fibronectin +/- transglutaminase, and then assessed by SDS-PAGE and Western blotting. WAP2 expression was investigated in human lung tissue sections via immunohistochemistry. Recombinant WAP2 inhibited cathepsin G activity. Monocytic cells pre-treated with rWAP2 before LPS stimulation showed significantly lower levels of IL-8 and MCP-1 production compared to cells stimulated with LPS alone. Recombinant WAP2 was conjugated to fibronectin in a transglutaminase-mediated reaction and retained antiprotease activity. WAP2 was detected in human lung tissue sections via immunohistochemistry. Together these results suggest a role for WAP2 in innate host defence. Further characterisation is required to fully understand the implications of WAP2 properties and its clinical significance.