title: Nonantibiotic macrolides restore airway macrophage phagocytic function with potential anti-inflammatory effects in chronic lung diseases.
abstract: We reported defective efferocytosis associated with cigarette smoking and/or airway inflammation in chronic lung diseases, including chronic obstructive pulmonary disease, severe asthma, and childhood bronchiectasis. We also showed defects in phagocytosis of nontypeableHaemophilus influenzae(NTHi), a common colonizer of the lower airway in these diseases. These defects could be substantially overcome with low-dose azithromycin; however, chronic use may induce bacterial resistance. The aim of the present study was therefore to investigate two novel macrolides-2'-desoxy-9-(S)-erythromycylamine (GS-459755) and azithromycin-based 2'-desoxy molecule (GS-560660)-with significantly diminished antibiotic activity againstStaphylococcus aureus,Streptococcus pneumonia,Moraxella catarrhalis, andH. influenzaeWe tested their effects on efferocytosis, phagocytosis of NTHi, cell viability, receptors involved in recognition of apoptotic cells and/or NTHi (flow cytometry), secreted and cleaved intracellular IL-1β (cytometric bead array, immunofluorescence/confocal microscopy), and nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) using primary alveolar macrophages and THP-1 macrophages ± 10% cigarette smoke extract. Dose-response experiments showed optimal prophagocytic effects of GS-459755 and GS-560660 at concentrations of 0.5-1 µg/ml compared with our findings with azithromycin. Both macrolides significantly improved phagocytosis of apoptotic cells and NTHi (e.g., increases in efferocytosis and phagocytosis of NTHi: GS-459755, 23 and 22.5%,P= 0.043; GS-560660, 23.5 and 22%,P= 0.043, respectively). Macrophage viability remained >85% following 24 h exposure to either macrolide at concentrations up to 20 µg/ml. Secreted and intracellular-cleaved IL-1β was decreased with both macrolides with no significant changes in recognition molecules c-mer proto-oncogene tyrosine kinase; scavenger receptor class A, member 1; Toll-like receptor 2/4; or CD36. Particulate cytoplasmic immunofluorescence of NLRP3 inflammasome was also reduced significantly. We conclude that GS-459755 and GS-560660 may be useful for reducing airway inflammation in chronic lung diseases without inducing bacterial resistance.
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia; sandra.hodge@sa.gov.au.|Department of Medicine, University of Adelaide, Adelaide, Australia; and.
authors.forename: Sandra
authors.initials: S
authors.lastname: Hodge
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.
authors.forename: Hai B
authors.initials: HB
authors.lastname: Tran
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.
authors.forename: Rhys
authors.initials: R
authors.lastname: Hamon
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.|Department of Medicine, University of Adelaide, Adelaide, Australia; and.
authors.forename: Eugene
authors.initials: E
authors.lastname: Roscioli
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.|Department of Medicine, University of Adelaide, Adelaide, Australia; and.
authors.forename: Greg
authors.initials: G
authors.lastname: Hodge
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.|Department of Medicine, University of Adelaide, Adelaide, Australia; and.
authors.forename: Hubertus
authors.initials: H
authors.lastname: Jersmann
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.|Department of Medicine, University of Adelaide, Adelaide, Australia; and.
authors.forename: Miranda
authors.initials: M
authors.lastname: Ween
authors.affiliation: Lung Research Unit, Hanson Institute, and Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, Australia.|Department of Medicine, University of Adelaide, Adelaide, Australia; and.
authors.forename: Paul N
authors.initials: PN
authors.lastname: Reynolds
authors.affiliation: Gilead Sciences, Seattle, Washington.
authors.forename: Arthur
authors.initials: A
authors.lastname: Yeung
authors.affiliation: Gilead Sciences, Seattle, Washington.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Treiberg
authors.affiliation: Gilead Sciences, Seattle, Washington.
authors.forename: Sibylle
authors.initials: S
authors.lastname: Wilbert
mesh_terms: D000328:Adult; D000900:Anti-Bacterial Agents / Q000494:pharmacology / Q000627:therapeutic use*; D000893:Anti-Inflammatory Agents / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002908:Chronic Disease; D004917:Erythromycin / Q000031:analogs & derivatives / Q000494:pharmacology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006193:Haemophilus influenzae / Q000187:drug effects; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D008168:Lung / Q000473:pathology*; D008171:Lung Diseases / Q000188:drug therapy* / Q000382:microbiology / Q000473:pathology; D018942:Macrolides / Q000494:pharmacology / Q000627:therapeutic use*; D016676:Macrophages, Alveolar / Q000473:pathology*; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000378:metabolism; D010587:Phagocytosis / Q000187:drug effects*; D000090063:Proto-Oncogene Mas; D011956:Receptors, Cell Surface / Q000378:metabolism; D012907:Smoking / Q000009:adverse effects
chemical_list: C582729:2'-desoxy-9-erythromycylamine; D000900:Anti-Bacterial Agents; D000893:Anti-Inflammatory Agents; D053583:Interleukin-1beta; C000715470:MAS1 protein, human; D018942:Macrolides; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D000090063:Proto-Oncogene Mas; D011956:Receptors, Cell Surface; D004917:Erythromycin
keywords: azithromycin; chronic lung disease; chronic obstructive pulmonary disease; nonantibiotic macrolides; nontypeable H. influenzae