[en] Secreted deoxyribonucleases (DNases), such as DNase-I and DNase-IL3, degrade extracellular DNA, and endogenous DNases have roles in resolving airway inflammation and guarding against autoimmune responses to nucleotides. Subsets of patients with asthma have high airway DNA levels, but information about DNase activity in health and in asthma is lacking. To characterize DNase activity in health and in asthma, we developed a novel kinetic assay using a Taqman probe sequence that is quickly cleaved by DNase-I to produce a large product signal. We used this kinetic assay to measure DNase activity in sputum from participants in the Severe Asthma Research Program (SARP)-3 (n = 439) and from healthy controls (n = 89). We found that DNase activity was lower than normal in asthma [78.7 relative fluorescence units (RFU)/min vs. 120.4 RFU/min, P < 0.0001]. Compared to patients with asthma with sputum DNase activity in the upper tertile activity levels, those in the lower tertile of sputum DNase activity were characterized clinically by more severe disease and pathologically by airway eosinophilia and airway mucus plugging. Carbamylation of DNase-I, a post-translational modification that can be mediated by eosinophil peroxidase, inactivated DNase-I. In summary, a Taqman probe-based DNase activity assay uncovers low DNase activity in the asthma airway that is associated with more severe disease and airway mucus plugging and may be caused, at least in part, by eosinophil-mediated carbamylation.NEW & NOTEWORTHY We developed a new DNase assay and used it to show that DNase activity is impaired in asthma airways.
Disciplines :
Life sciences: Multidisciplinary, general & others
Author, co-author :
Charbit, Annabelle R ; Cardiovascular Research Institute (CVRI), University of California, San Francisco, California, United States
Liégeois, Maude ; Université de Liège - ULiège > Département des sciences fonctionnelles (DSF) > Biochimie et biologie moléculaire ; Cardiovascular Research Institute (CVRI), University of California, San Francisco, California, United States
Raymond, Wilfred W; Cardiovascular Research Institute (CVRI), University of California, San Francisco, California, United States
Comhair, Suzy A A; Inflammation and Immunity, Cleveland Clinic, Cleveland, Ohio, United States
Johansson, Mats W ; Pulmonary and Critical Care Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States
Hastie, Annette T ; Pulmonary, Critical Care, Allergy and Immunologic Diseases, Wake Forest University, Winston-Salem, North Carolina, United States
Bleecker, Eugene R; Asthma and Airway Disease Research Center, University of Arizona, Phoenix, Arizona, United States
Fajt, Merritt; Pulmonary, Allergy and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
Castro, Mario ; Pulmonary, Critical Care and Sleep Medicine, University of Kansas, Kansas City, Kansas, United States
Sumino, Kaharu; Division of Pulmonary and Critical Care Medicine, Washington University in St. Louis, St. Louis, Missouri, United States
Erzurum, Serpil C ; Inflammation and Immunity, Cleveland Clinic, Cleveland, Ohio, United States
Israel, Elliot ; Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, Massachusetts, United States
Jarjour, Nizar N; Pulmonary and Critical Care Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States
Mauger, David T ; Division of Biostatistics and Bioinformatics, Penn State University, Hershey, Pennsylvania, United States
Moore, Wendy C; Pulmonary, Critical Care, Allergy and Immunologic Diseases, Wake Forest University, Winston-Salem, North Carolina, United States
Wenzel, Sally E ; Pulmonary, Allergy and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
Woodruff, Prescott G; Cardiovascular Research Institute (CVRI), University of California, San Francisco, California, United States
Levy, Bruce D ; Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, Massachusetts, United States
Tang, Monica C ; Cardiovascular Research Institute (CVRI), University of California, San Francisco, California, United States
Fahy, John V ; Cardiovascular Research Institute (CVRI), University of California, San Francisco, California, United States
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