title: Pulmonary surfactant protein a inhibits macrophage reactive oxygen intermediate production in response to stimuli by reducing NADPH oxidase activity.
abstract: Alveolar macrophages are important host defense cells in the human lung that continuously phagocytose environmental and infectious particles that invade the alveolar space. Alveolar macrophages are prototypical alternatively activated macrophages, with up-regulated innate immune receptor expression, down-regulated costimulatory molecule expression, and limited production of reactive oxygen intermediates (ROI) in response to stimuli. Surfactant protein A (SP-A) is an abundant protein in pulmonary surfactant that has been shown to alter several macrophage (Mphi) immune functions. Data regarding SP-A effects on ROI production are contradictory, and lacking with regard to human Mphi. In this study, we examined the effects of SP-A on the oxidative response of human Mphi to particulate and soluble stimuli using fluorescent and biochemical assays, as well as electron paramagnetic resonance spectroscopy. SP-A significantly reduced Mphi superoxide production in response to the phorbol ester PMA and to serum-opsonized zymosan (OpZy), independent of any effect by SP-A on zymosan phagocytosis. SP-A was not found to scavenge superoxide. We measured Mphi oxygen consumption in response to stimuli using a new oxygen-sensitive electron paramagnetic resonance probe to determine the effects of SP-A on NADPH oxidase activity. SP-A significantly decreased Mphi oxygen consumption in response to PMA and OpZy. Additionally, SP-A reduced the association of NADPH oxidase component p47(phox) with OpZy phagosomes as determined by confocal microscopy, suggesting that SP-A inhibits NADPH oxidase activity by altering oxidase assembly on phagosomal membranes. These data support an anti-inflammatory role for SP-A in pulmonary homeostasis by inhibiting Mphi production of ROI through a reduction in NADPH oxidase activity.
authors.affiliation: Interdisciplinary Graduate Program in Immunology, University of Iowa, Iowa City, IA 52240, USA.
authors.forename: Joy E
authors.initials: JE
authors.lastname: Crowther
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Kutala
authors.forename: Periannan
authors.initials: P
authors.lastname: Kuppusamy
authors.forename: J Scott
authors.initials: JS
authors.lastname: Ferguson
authors.forename: Alison A
authors.initials: AA
authors.lastname: Beharka
authors.forename: Jay L
authors.initials: JL
authors.lastname: Zweier
authors.forename: Francis X
authors.initials: FX
authors.lastname: McCormack
authors.forename: Larry S
authors.initials: LS
authors.lastname: Schlesinger
mesh_terms: D006801:Humans; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism; D019255:NADPH Oxidases / Q000502:physiology*; D010587:Phagocytosis / Q000187:drug effects; D010750:Phosphoproteins; D037662:Pulmonary Surfactant-Associated Protein A / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016897:Respiratory Burst / Q000187:drug effects; D013481:Superoxides / Q000378:metabolism
chemical_list: D010750:Phosphoproteins; D037662:Pulmonary Surfactant-Associated Protein A; D017382:Reactive Oxygen Species; D013481:Superoxides; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1