Der p 1 facilitates transepithelial allergen delivery by disruption of tight junctions
Hong Wan, … , Mark B. Cannell, Clive Robinson
Hong Wan, … , Mark B. Cannell, Clive Robinson
Published July 1, 1999
Citation Information: J Clin Invest. 1999;104(1):123-133. https://doi.org/10.1172/JCI5844.
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Article

Der p 1 facilitates transepithelial allergen delivery by disruption of tight junctions

Abstract

House dust mite (HDM) allergens are important factors in the increasing prevalence of asthma. The lung epithelium forms a barrier that allergens must cross before they can cause sensitization. However, the mechanisms involved are unknown. Here we show that the cysteine proteinase allergen Der p 1 from fecal pellets of the HDM Dermatophagoides pteronyssinus causes disruption of intercellular tight junctions (TJs), which are the principal components of the epithelial paracellular permeability barrier. In confluent airway epithelial cells, Der p 1 led to cleavage of the TJ adhesion protein occludin. Cleavage was attenuated by antipain, but not by inhibitors of serine, aspartic, or matrix metalloproteinases. Putative Der p 1 cleavage sites were found in peptides from an extracellular domain of occludin and in the TJ adhesion protein claudin-1. TJ breakdown nonspecifically increased epithelial permeability, allowing Der p 1 to cross the epithelial barrier. Thus, transepithelial movement of Der p 1 to dendritic antigen-presenting cells via the paracellular pathway may be promoted by the allergen’s own proteolytic activity. These results suggest that opening of TJs by environmental proteinases may be the initial step in the development of asthma to a variety of allergens.

Authors

Hong Wan, Helen L. Winton, Christian Soeller, Euan R. Tovey, Dieter C. Gruenert, Philip J. Thompson, Geoffrey A. Stewart, Graham W. Taylor, David R. Garrod, Mark B. Cannell, Clive Robinson

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Figure 8

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(a) Occludin immunoblots from MDCK cells. Lane 1: untreated cells; lane ...
(a) Occludin immunoblots from MDCK cells. Lane 1: untreated cells; lane 2: cells treated with heat-inactivated Der p 1 for 1 hour; lane 3: following 1-hour treatment with active Der p 1 (activity 17 nmol/min); lane 4: active Der p 1 with 100 μM antipain. Note attenuation of degradation by antipain or heat inactivation. (b) Inhibition of occludin degradation in MDCK cells by occludin peptide 88AWDRGYGTSLLG99. Lane 5: cells treated for 1 hour with Der p 1; lane 6: peptide (100 μM) and cells alone; lanes 7–9: cells treated with peptide and Der p 1 for 0.25, 0.5, and 1 hour, respectively. (c) Lack of effect of mixed proteinase inhibitors on occludin degradation in MDCK cells treated with Der p 1. Lane 10: untreated cells; lane 11: mixed inhibitors (100 μM AEBSF, 1 μM BB-250, and 1 μM pepstatin) alone; lane 12: Der p 1 alone; lane 13: Der p 1 with mixed inhibitors. (d) HPLC/electrospray–selected ion mass chromatograms for m/z 490.3 (M+H+, GTSLLG) and m/z 563.3 (M+H22+, AWDRGYGTSL) are shown following Der p 1 treatment of AWDRGYGTSLLG in the presence of E-64 (100 μM), antipain (100 μM), or the absence of inhibitors. Both inhibitors reduced the formation of 2 major enzymic fragments, AWDRGYGTSL and GTSLLG.