Abstract
Workers in swine confinement facilities develop chronic inflammatory lung disease, and an aqueous hog‐barn dust extract (HDE) mimics disease‐relevant effects in cultured cells and animal models. We hypothesized that multiple components in HDE contribute to its effects on airway cells. The sensitivity/resistance of HDE activities for IL6/8 release and EGF receptor (EGFR) modulation in BEAS‐2B airway epithelial cells to chemical and enzyme treatments suggested heat‐stable proteins as mediators. Gel filtration revealed three separate peaks of activity for IL6/8 release, one >100 kDa, one ~30–50 kDa, and one of very low MW. Fractions of ~3–10 kDa had activity to phosphorylate and decrease binding to EGFRs. Mass spectrometry analysis of proteins in active fractions identified porcine pancreatic elastase (PPE). PPE mimicked some but not all cellular effects of complete HDE. Zymography identified protease activity in HDE, and protease inhibitors markedly reduced HDE activity on cells. Hog manure extract also exhibited activity on IL6/8 release and EGFRs. Thus multiple factors contribute to HDE effects, and manure‐derived PPE and/or other proteases are candidate mediators of hog‐barn dust‐induced lung disease. Supported by NIH R01‐OH008539.
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