Volume 6 Supplement 1

2nd WAO International Scientific Conference (WISC 2012), Abstracts

Open Access

Mechanisims of asthma and allergic disease – 1087. Serine protease activity of per a 10 is requried to activiate airway epithelial cells

  • Sagar Laxman Kale1 and
  • Naveen Arora1
World Allergy Organization Journal20136(Suppl 1):P83

DOI: 10.1186/1939-4551-6-S1-P83

Published: 23 April 2013

Background

Airway epithelial cells play an important role in initiating airway allergic responses by secreting a variety of proinflammatory cytokines. The present study was aimed to elucidate the effect of serine protease activity of Per a 10 on biochemical properties of an airway epithelial cell line.

Methods

Alveolar type II epithelial cell line A549 was incubated with active (nPer a 10), AEBSF inactivated (iPer a 10), heat inactivated (ΔPer a 10) native Per a 10 and inactive recombinant Per a 10 (rPer a 10). After incubation pro-inflammatory cytokines IL-6, IL-8 and GMCSF were measured by ELISA. Intracellular Ca2+ mobilisation was determined in A549 cells by Fluo 4AM. PAR-2 derived peptide (GTNRSSKGRSLIGKVDGTSHVTGKGVTC) was incubated with nPer a 10 and resultant cleaved products were subjected to LC-MS.

Results

nPer a 10 caused desquamation of cells that led to the secretion of pro-inflammatory cytokines from A549 cells in concentration dependent manner. ΔPer a 10 whose activity was abolished by heat treatment and rPer a 10 failed to secrete any of these cytokines. iPer a 10 showed some residual activity after treating with AEBSF and caused lower but significant secretion of proinflammatory cytokines. Per a 10 was able to mobilise Ca2+ in a concentration and activity dependent manner. nPer a 10 cleaved the PAR-2 derived peptide (GTNRSSKGRSLIGKVDGTSHVTGKGVTC) between arginine and serine residues as determined by LC-MS, to expose the PAR-2 specific ligand SLIGKV. Whereas rPer a 10 and inactive Per a 10 failed both to mobilise Ca2+ and to cleave PAR-2 derived peptide. Thus Per a 10 activated epithelial cells to secrete proinflammatory cytokines in activity dependent manner via PAR-2 receptors.

Conclusions

Per a 10 activates epithelial cells to secrete proinflammatory cytokines that mediate the allergic responses and the activation is through the cleavage of PAR-2 receptors.

Authors’ Affiliations

(1)
Allergy and Immunology, Institute of Genomics and Integrative Biology

Copyright

© Kale and Arora; licensee BioMed Central Ltd. 2013

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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