title: Eosinophil protein in airway macrophages: a novel biomarker of eosinophilic inflammation in patients with asthma.
abstract: 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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.|Children's Community Health Service, Leicester City NHS Primary Care Trust.
authors.forename: Neeta S
authors.initials: NS
authors.lastname: Kulkarni
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Fay
authors.initials: F
authors.lastname: Hollins
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Amanda
authors.initials: A
authors.lastname: Sutcliffe
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Ruth
authors.initials: R
authors.lastname: Saunders
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Sachil
authors.initials: S
authors.lastname: Shah
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Salman
authors.initials: S
authors.lastname: Siddiqui
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Sumit
authors.initials: S
authors.lastname: Gupta
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Pranab
authors.initials: P
authors.lastname: Haldar
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Ruth
authors.initials: R
authors.lastname: Green
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Ian
authors.initials: I
authors.lastname: Pavord
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wardlaw
authors.affiliation: Institute for Lung Health, Glenfield Hospital, Leicester, and the Department of Infection, Immunity and Inflammation, University of Leicester, United Kingdom.
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Brightling
mesh_terms: D000305:Adrenal Cortex Hormones / Q000627:therapeutic use; D000328:Adult; D017209:Apoptosis; D001249:Asthma / Q000150:complications* / Q000188:drug therapy / Q000378:metabolism; D015415:Biomarkers; D003430:Cross-Sectional Studies; D047150:Eosinophil Cationic Protein / Q000032:analysis*; D047209:Eosinophil Peroxidase / Q000032:analysis*; D004802:Eosinophilia / Q000175:diagnosis*; D005260:Female; D006801:Humans; D008264:Macrophages / Q000737:chemistry*; D008297:Male; D008875:Middle Aged; D013183:Sputum / Q000737:chemistry / Q000166:cytology
chemical_list: D000305:Adrenal Cortex Hormones; D015415:Biomarkers; D047209:Eosinophil Peroxidase; D047150:Eosinophil Cationic Protein; C479220:RNASE3 protein, human