Purification and Characterization of a Serine Protease from Organic Dust and Elucidation of Its Inductive Effects on Lung Inflammatory Mediators

Details

• Personal Author:
  Boggaram V ; Keshava S ; Kusampudi S ; Meganathan V
• Description:
  Organic dust inhalation is associated with the development of respiratory diseases. Serine protease activities in organic dusts were previously reported to contribute to the induction of lung inflammatory mediators however, the identities of the proteases and the mechanisms by which they induce inflammatory mediators are unknown. The goal of this study was to purify and characterize serine protease(s) from organic dust and elucidate mechanisms by which they induce lung inflammatory mediators. A serine protease was purified from poultry organic dust by benzamidine-agarose affinity chromatography. Mass spectrometry and amino-terminal sequence analysis identified the purified protease as chicken trypsin II P29. Purified protease induced proinflammatory cytokine levels in Beas2B and NHBE epithelial and THP-1 macrophage cells. Treatment with the purified protease increased cellular and mitochondrial reactive oxygen species (ROS) generation. Induction of inflammatory mediators and ROS were suppressed by serine protease inhibitors and antioxidants. Purified protease activated PKC, MAPK1/3 and MAPK14 signaling and NF-kB and Stat-3, and chemical inhibitors targeting these pathways suppressed induction of inflammatory mediators. Calcium mobilization studies showed that the purified protease activated protease activated receptors (PAR) F2R, F2RL1, F2RL2, F2RL3, and F2R and F2RL1 knockdown suppressed the induction of inflammatory mediators. Intranasal instillation of purified protease increased lung CXCL1, IL-6 and TNF levels in mice. Our studies have shown that chicken trypsin is a proinflammatory constituent of poultry organic dust, and induces lung inflammatory mediators via increased ROS and PAR activation in a cell signaling pathway involving PKC, MAPK1/3 and MAPK14 and NF-kB and Stat-3. [Description provided by NIOSH]
• Subjects:
  Agriculture Chromatography Dust Food Industry Immune System Lung Occupational Health Respiratory System Respiratory Tract Diseases Safety Trypsin

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• Keywords:
  Author Keywords: Agricultural Dust Cytokines Gene Regulation Immune Reaction Mass Spectrometry Organic Dusts Poultry Reactive Oxygen Species Respiratory Diseases Trypsin
• ISSN:
  1040-0605
• Document Type:
  Text
• Funding:
  Cooperative Agreement
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 6 ; Texas
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  325
• Issue:
  1
• NIOSHTIC Number:
  nn:20067681
• Citation:
  Am J Physiol Lung Cell Mol Physiol 2023 Jul; 325(1):L74-L90
• Contact Point Address:
  Vijay Boggaram, Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United States
• Email:
  vijay.boggaram@uthct.edu
• Federal Fiscal Year:
  2023
• NORA Priority Area:
  Agriculture, Forestry and Fishing
• Performing Organization:
  University of Texas Health Center at Tyler
• Peer Reviewed:
  True
• Start Date:
  20010930
• Source Full Name:
  American Journal of Physiology: Lung Cellular and Molecular Physiology
• End Date:
  20270929
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:c411a71b4302a05a0fc9f53e0d4d1112fb10391d97c1baf02e5fc919540271a07e05f040cc8afe929e5a4c1d0954ff55ad73481f5e56070b16fa497d286e4200
• Download URL:
  https://stacks.cdc.gov/view/cdc/229949/cdc_229949_DS1.pdf
• File Type:
  [PDF - 4.10 MB ]