title: Mechanisms of suppression of alveolar epithelial cell GM-CSF expression in the setting of hyperoxic stress.
abstract: Pulmonary expression of granulocyte/macrophage colony-stimulating factor (GM-CSF) is critically important for normal functional maturation of alveolar macrophages. We found previously that lung GM-CSF is dramatically suppressed in mice exposed to hyperoxia. Alveolar epithelial cells (AEC) are a major source of GM-CSF in the peripheral lung, and in vivo hyperoxia resulted in greatly reduced expression of GM-CSF protein by AEC ex vivo. We now explore the mechanisms responsible for this effect, using primary cultures of murine AEC exposed to hyperoxia in vitro. Exposure of AEC to 80% oxygen/5% CO(2) for 48 h did not induce overt toxicity, but resulted in significantly decreased GM-CSF protein and mRNA expression compared with cells in normoxia. Similar effects were seen when AEC were stressed with serum deprivation, an alternative inducer of oxidative stress. The effects in AEC were opposite those in a murine lung epithelial cell line (MLE-12 cells), in which hyperoxia induced GM-CSF expression. Both hyperoxia and serum deprivation resulted in increased intracellular reactive oxygen species (ROS) in AEC. Hyperoxia and serum deprivation induced significantly accelerated turnover of GM-CSF mRNA. Treatment of AEC with catalase during oxidative stress preserved GM-CSF protein and mRNA and was associated with stabilization of GM-CSF mRNA. We conclude that hyperoxia-induced suppression of AEC GM-CSF expression is a function of ROS-induced destabilization of GM-CSF mRNA. We speculate that AEC oxidative stress results in significantly impaired pulmonary innate immune defense due to effects on local GM-CSF expression in the lung.
authors.affiliation: Department of Veterans Affairs Medicine Center, and Division of Respiratory, Critical Care, and Occupational Pulmonary Medicine, University of Utah School of Medicine, 26 North 1900 East, Salt Lake City, UT 84132, USA.
authors.forename: Anne
authors.initials: A
authors.lastname: Sturrock
authors.forename: Timothy
authors.initials: T
authors.lastname: Vollbrecht
authors.forename: Mustafa
authors.initials: M
authors.lastname: Mir-Kasimov
authors.forename: Michael
authors.initials: M
authors.lastname: McManus
authors.forename: Steven E
authors.initials: SE
authors.lastname: Wilcoxen
authors.forename: Robert
authors.initials: R
authors.lastname: Paine
mesh_terms: D056809:Alveolar Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D002374:Catalase / Q000494:pharmacology; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D016895:Culture Media, Serum-Free; D005786:Gene Expression Regulation / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism*; D018496:Hyperoxia / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D016895:Culture Media, Serum-Free; D012333:RNA, Messenger; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D002374:Catalase