Detection of Intracellular Markers in Airway Inflammation a Biochemical and Immunocytochemical Study in Induced Sputum

6 ABBREVIATIONS APAAP alkaline phosphatase-anti-alkaline phosphatase AUC area under curve BAL bronchoalveolar lavage BSA bovine serum albumin BUD budesonide COPD chronic obstructive pulmonary disease CTAB N-cetyl-N,N,N,-trimethylammonium bromide DTE dithioerythritol DTT dithiothreitol EG2, EG1 monoclonal antibodies against ECP and/or EPX/EDN ECP eosinophil cationic protein EPO eosinophil peroxidase EPX/EDN eosinophil protein X/eosinophil-derived neurotoxin FA formaldehyde FEV 1 forced expiratory volume in one second GM-CSF granulocyte-macrophage colony-stimulating factor HNL human neutrophil lipocalin HSA human serum albumin IgG, IgE immunoglobulin G, immunoglobulin E IL-8 interleukin-8 kDa kiloDalton MBP major basic protein MGG May-Grünwald-Giemsa MPO myeloperoxidase NaCl sodium chloride OPF Ortho Permeafi x PAF platelet activating factor PBS phosphate-buffered saline PD 15 provocative dose of histamine causing a 15% reduction in FEV 1 (mg of histamine diphosphate) PD 20 FEV 1 provocative dose of histamine causing a 20% reduction in FEV 1 (mg of histamine diphosphate) PEF peak expiratory fl ow PFA paraformaldehyde PLP periodate-lysine-paraformaldehyde ROC receiver-operating characteristic SD standard deviation TER terbutaline 7 ORIGINAL PUBLICATIONS This thesis is based on the following original publications, referred to in the text by the Roman numeral concerned. I Metso T, Rytilä P, Peterson C, Haahtela T. Granulocyte markers in induced sputum in patients with respiratory disorders and healthy persons obtained by two sputum-processing methods. Cell-specifi c markers for eosi-nophils and neutrophils in sputum and bronchoalveolar lavage of patients with respiratory conditions and healthy persons. Eosinophil cationic protein in induced sputum as a marker of infl ammation in asthmatic children. Some unpublished results are also reported and discussed. 8 ABSTRACT Characteristic features in asthma and chronic obstructive pulmonary disease (COPD) are airfl ow limitation and bronchial infl ammation. During airway infl ammation, increased numbers of activated cells (e.g. eosinophils and neutrophils) associated with infl ammation are found in bronchial mucosa, in bronchoalveolar lavage (BAL) fl uid, in sputum, and, in severe cases, in blood. Both eosinophils and neutrophils contain various mediators in their intracellular granules. When such cells are activated, various types of harmful mediator are released into the immediate environment. Until recently, investigation of airway infl ammation has mainly relied on monitoring of lung function. However, numerous bronchial biopsy and BAL fl uid studies have revealed that airway infl ammation can occur in the absence of bronchial hyper-reactivity, if respiratory disease is mild or clinically well controlled. Cellular and biochemical accompaniments of airway infl ammation have been widely studied by investigation of BAL fl uid and bronchial …