Eosinophil protein in airway macrophages: a novel biomarker of eosinophilic inflammation in patients with asthma.

BACKGROUND: Noneosinophilic asthma is common across asthma severities. However, in patients with moderate-to-severe disease, the absence of sputum eosinophilia cannot distinguish between asthmatic subjects with eosinophilic inflammation controlled by corticosteroids versus those in whom eosinophilic inflammation is not a component of the disease. OBJECTIVES: We sought to develop a method to quantify eosinophil proteins in airway macrophages as a novel biomarker of eosinophilic airway inflammation. METHODS: Eosinophil proteins in airway macrophages were assessed by means of flow cytometry, immunofluorescence, and cytoplasmic hue change after ingestion of apoptotic eosinophils. Airway macrophage median percentage of red-hued area in stained sputum cytospin preparations was assessed by means of image analysis from (1) subjects with mild-to-severe asthma, subjects with nonasthmatic eosinophilic bronchitis, and healthy control subjects; (2) subjects with eosinophilic severe asthma after treatment with prednisolone; and (3) subject with noneosinophilic asthma before corticosteroid withdrawal. RESULTS: Eosinophil proteins were detected in airway macrophages, and cytoplasmic red hue increased after ingestion of apoptotic eosinophils. Airway macrophage percentage redhued area was increased in subjects with moderate-to-severe asthma compared with that seen in subjects with mild asthma and healthy control subjects, was similar in those with or without a sputum eosinophilia, and was increased after corticosteroid therapy. In asthmatic subjects without sputum eosinophilia, the airway macrophage percentage red-hued area was increased in subjects who did versus those who did not have sputum eosinophilia after corticosteroid withdrawal. CONCLUSIONS: Eosinophil proteins can be reliably measured in airway macrophages. In combination with sputum eosinophilia, the macrophage eosinophil protein content might further define the asthma phenotype and provide an additional tool to direct therapy.

Domestic smoke exposure is associated with alveolar macrophage particulate load.

OBJECTIVES: Indoor air pollution is associated with impaired respiratory health. The pre-dominant indoor air pollutant to which two billion of the world's population is exposed is biomass fuel smoke. We tested the hypothesis that reported smoke exposure in men and women is associated with increased alveolar macrophage uptake of biomass smoke particulates. METHODS: Healthy volunteers attending for research bronchoscopy in Malawi completed a questionnaire assessment of smoke exposure. Particulate matter visible in alveolar macrophages (AM) was quantified using digital image analysis. The geometric mean of the percentage area of the cytoplasm occupied by particulates in 50 cover-slip adherent AM was calculated and termed particulate load. RESULTS: In 57 subjects (40 men and 17 women) there was a significant difference between the particulate load in groups divided according to pre-dominant lighting form used at home (ANOVA P = 0.0009) and type of cooking fuel (P = 0.0078). CONCLUSIONS: Particulate load observed in macrophages is associated with the reported type of biomass fuel exposure. Macrophage function in relation to respiratory health should now be investigated in biomass smoke exposed subjects.

Carbon loading of alveolar macrophages in adults and children exposed to biomass smoke particles.

Exposure to carbonaceous particles from biomass burning is associated with increased respiratory morbidity in both women and children in the developing world. However, the amount of carbon reaching lower airway cells has not been determined in these populations. Alveolar macrophages (AM) remove inhaled particulate matter (PM), and are implicated in the pathogenesis of PM-induced lung disease. In this study, we aimed to compare AM carbon loading in women and children exposed to biomass PM in Gondar, Ethiopia, with individuals exposed to fossil-fuel PM in the developed world (Leicester, UK). To achieve these aims, we sampled AM from Ethiopian mothers and children, and from UK adults and children using induced sputum (IS). AM were imaged under light microscopy, and the total two-dimensional surface area of carbon within each AM determined by image analysis. AM containing carbon were detected in all subjects. The total surface area of carbon per AM was higher in Ethiopian women (n=10) compared with UK adults (n=10, median 9.19 vs. 0.71 microm2/AM, p=0.0002). Similarly, the total surface area of carbon per AM was higher in Ethiopian children (n=10) compared with UK children (n=10, 3.32 vs. 0.44 microm2/AM, p=0.0002). However, loading in Ethiopian children was lower than paired maternal levels (3.32 vs. 9.19 microm2/AM, p=0.011). We conclude that analysis of AM obtained by induced sputum is a practical way of quantifying natural exposure of the lower airway to carbonaceous particles from the burning of biomass fuels.