Surfactant protein-D modulation of pulmonary macrophage phenotype is controlled by S -nitrosylation

• Published in: American journal of physiology. Lung cellular and molecular physiology Vol. 317; no. 5; pp. L539 - L549
• Main Authors: Guo, Chang-Jiang, Atochina-Vasserman, Elena N, Abramova, Elena, Smith, Ley Cody, Beers, Michael F, Gow, Andrew J
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.11.2019
• Series: Translational Physiology
• Subjects: Alveoli; Animals; Bronchoalveolar Lavage Fluid - chemistry; Bronchoalveolar Lavage Fluid - immunology; Bronchus; CCL20 protein; Cellular manufacture; Chemokine CCL20 - genetics; Chemokine CCL20 - immunology; Chemokine CXCL1 - genetics; Chemokine CXCL1 - immunology; Cyclooxygenase 2 - genetics; Cyclooxygenase 2 - immunology; Female; Gene expression; Genes; Immunity, Innate; Inflammation; Innate immunity; Interleukin 1; Interleukin-1beta - genetics; Interleukin-1beta - immunology; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Lung - immunology; Lung - metabolism; Lung - microbiology; Macrophages; Macrophages, Alveolar - drug effects; Macrophages, Alveolar - immunology; Macrophages, Alveolar - microbiology; Male; Metabolites; Mice; Mice, Inbred C57BL; NF-kappa B - genetics; NF-kappa B - immunology; NF-κB protein; Nitric Oxide Synthase Type II - genetics; Nitric Oxide Synthase Type II - immunology; Nitric-oxide synthase; Nitroso Compounds - immunology; Nitroso Compounds - metabolism; Phenotype; Phenotypes; Pneumocystis - growth & development; Pneumocystis - pathogenicity; Pneumocystis Infections - genetics; Pneumocystis Infections - immunology; Pneumocystis Infections - metabolism; Pneumocystis Infections - microbiology; Protein Processing, Post-Translational; Proteins; Pulmonary Surfactant-Associated Protein D - genetics; Pulmonary Surfactant-Associated Protein D - immunology; Pulmonary Surfactant-Associated Protein D - metabolism; RAW 264.7 Cells; Surfactants; Target recognition; Tumor necrosis factor; Tumor Necrosis Factor-alpha - genetics; Tumor Necrosis Factor-alpha - immunology; Vascular Cell Adhesion Molecule-1 - genetics; Vascular Cell Adhesion Molecule-1 - immunology; surfactant protein-D; S-nitrosothiol; macrophage phenotype; pneumocystis pneumonia; inducible nitric oxide synthase
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/ajplung.00506.2018

Surfactant protein-D (SP-D) is a regulator of pulmonary innate immunity whose oligomeric state can be altered through -nitrosylation to regulate its signaling function in macrophages. Here, we examined how nitrosylation of SP-D alters the phenotypic response of macrophages to stimuli both in vivo and in vitro. Bronchoalveolar lavage (BAL) from C57BL6/J and SP-D-overexpressing (SP-D OE) mice was incubated with RAW264.7 cells ± LPS. LPS induces the expression of the inflammatory genes and , which is reduced 10-fold by SP-D OE-BAL. -nitrosylation of the SP-D OE-BAL (SNO-SP-D OE-BAL) abrogated this inhibition. SNO-SP-D OE-BAL alone induced and expression. PCR array analysis of macrophages incubated with SP-D OE-BAL (±LPS) shows increased expression of repair genes, , , and , that was accentuated by LPS. LPS increases inflammatory gene expression, , , , and , which was accentuated by SNO-SP-D OE-BAL but inhibited by SP-D OE-BAL. The transcription factor NF-κB was identified as a target for SNO-SP-D by IPA, which was confirmed by Trans-AM ELISA in vitro. In vivo, SP-D overexpression increases the burden of infection in a model while increasing cellular recruitment. Expression of iNOS and the production of NO metabolites were significantly reduced in SP-D OE mice relative to C57BL6/J. Inflammatory gene expression was increased in infected C57BL6/J mice but decreased in SP-D OE. SP-D oligomeric structure was disrupted in C57BL6/J infected mice but unaltered within SP-D OE. Thus SP-D modulates macrophage phenotype and the balance of multimeric to trimeric SP-D is critical to this regulation.