Neutrophils are a major source of the epithelial barrier disrupting cytokine oncostatin M in patients with mucosal airways disease

Abstract

We have previously shown that oncostatin M (OSM) levels are increased in nasal polyps (NPs) of patients with chronic rhinosinusitis (CRS), as well as in bronchoalveolar lavage fluid, after segmental allergen challenge in allergic asthmatic patients. We also showed invitro that physiologic levels of OSM impair barrier function in differentiated airway epithelium. We sought to determine which hematopoietic or resident cell type or types were the source of the OSM expressed in patients with mucosal airways disease. Paraffin-embedded NP sections were stained with fluorescence-labeled specific antibodies against OSM, GM-CSF, and hematopoietic cell-specific markers. Live cells were isolated from NPs and matched blood samples for flow cytometric analysis. Neutrophils were isolated from whole blood and cultured with the known OSM inducers GM-CSF and follistatin-like 1, and OSM levels were measured in the supernatants. Bronchial biopsy sections from control subjects, patients with moderate asthma, and patients with severe asthma were stained for OSM and neutrophil elastase. OSM staining was observed in NPs, showed colocalization with neutrophil elastase (n=10), and did not colocalize with markers for eosinophils, macrophages, T cells, or B cells (n=3-5). Flow cytometric analysis of NPs (n=9) showed that 5.1%±2% of CD45+ cells were OSM+, and of the OSM+ cells, 56%±7% were CD16+Siglec-8-, indicating neutrophil lineage. Only 0.6 ±0.4% of CD45+ events from matched blood samples (n=5) were OSM+, suggesting that increased OSM levels in patients with CRS was locally stimulated and produced. Amajority of OSM+ neutrophils expressed arginase 1 (72.5%±12%), suggesting an N2 phenotype. GM-CSF levels were increased in NPs compared with those in control tissue and were sufficient to induce OSM production (P<.001) in peripheral blood neutrophils invitro. OSM+ neutrophils were also observed at increased levels in biopsy specimens from patients with severe asthma. Additionally, OSM protein levels were increased in induced sputum from asthmatic patients compared with that from control subjects (P<.05). Neutrophils are a major source of OSM-producing cells in patients with CRS and severe asthma.