Methods

Patients were admitted to the hospital with CAP (n= 11) and recruited from the emergency department after they provided informed consent. The inclusion criteria were radiological evidence of pneumonia, a clinical diagnosis of pneumonia using confusion, urea, respiratory rate, blood pressure, age> 65 years (CURB-65) score≥ 2, and age> 18 years. AAT glycoform patterns (M0–M8) were visualized using a Sebia Hydragel A1AT Isofocusing kit (2). AAT glycoanalysis was performed using liquid chromatography and exoglycosidase digestions (6). Plasma levels of IL-8 were determined by ELISA (R&D Systems) using a Spectra Max M3 plate reader. AAT was purified by affinity chromatography using Alpha-1 Antitrypsin Select resin (GE Healthcare)(2). Peripheral blood neutrophils were isolated by dextran sedimentation and Lymphoprep (Axis-Shield PoC) centrifugation, and modulation of IL-8 receptor engagement and neutrophil chemotaxis was as previously described (3).