Article Text

Neutrophil apoptosis, proinflammatory mediators and cell counts in bronchiectasis
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  1. A P Watt,
  2. V Brown,
  3. J Courtney,
  4. M Kelly,
  5. L Garske,
  6. J S Elborn,
  7. M Ennis
  1. Respiratory Research Group, Centre for Infection, Inflammation and Repair, The Queen’s University of Belfast, Belfast, UK
  1. Correspondence to:
    Dr A P Watt
    Respiratory Research Group, Centre for Infection, Inflammation and Repair, The Queen’s University of Belfast, Belfast, UK; a.wattqub.ac.uk

Abstract

Background: Lower airway secretions from patients with bronchiectasis show inflammatory cell infiltration and increased proinflammatory mediators. The aim of this study was to investigate the effects of antibiotic treatment for exacerbations on neutrophil apoptosis and necrosis.

Methods: Sputum was induced from 15 subjects with idiopathic bronchiectasis at the beginning of an acute exacerbation and after intravenous antibiotic treatment. Neutrophil apoptosis and necrosis were assessed using flow cytometry and morphology and the supernatant was analysed for concentrations of inflammatory mediators.

Results: Neutrophil numbers (×106 cells/g sputum) in sputum were significantly greater on day 0 than on day 14 (median difference (95% confidence interval (CI)) 5.14 (1.27 to 8.46), p = 0.02). Controls had a significantly higher percentage of sputum macrophages than patients with bronchiectasis (day 0, 1.35 (95% CI 0.48 to 2.89), p = 0.004; day 14, 1.09 (95% CI 0.26 to 2.86), p = 0.02). The concentrations of tumour necrosis factor α (pg/ml), interleukin 8 (ng/ml), and neutrophil elastase (ng/ml) in sputum supernatant were significantly reduced on day 14 compared with day 0 (median difference −94 (95% CI −158 to −27), p = 0.005; −106 (95% CI −189 to −50), p = 0.0006; and −73 451 (95% CI −135 495 to −12 303), p = 0.02 respectively). Patients with bronchiectasis had a significantly lower percentage of cells which were neither apoptotic nor necrotic than healthy controls (both days, −38.8 (95% CI −49.6 to −8.5), p = 0.002; −45.0 (95% CI −58.0 to −34.1), p = 0.0003, respectively), and on day 14 they had a significantly higher percentage of secondary necrotic cells than healthy controls (40 (95% CI 11.6 to 57.5), p = 0.004).

Conclusions: This study shows that antibiotic treatment affects concentrations of proinflammatory mediators and cell death and clearance may be altered in bronchiectasis.

  • induced sputum
  • inflammation
  • flow cytometry
  • CRP, C reactive protein
  • FEV1, forced expiratory volume in 1 second
  • IL, interleukin
  • NE, neutrophil elastase
  • TNFα, tumour necrosis factor α

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Supplementary materials

  • Web-only Figures

    Files in this Data Supplement:

    • [View Figure] - Figure S1 DiffQuik stain of sputum from a patient with bronchiectasis (�e100) showing an apoptotic neutrophil with condensed nuclei (closed arrow) and normal neutrophil.
    • [View Figure] - Figure S2(A) Dot plot (forward scatter versus side scatter) illustrating the position of the granulocyte population in sputum (circled) induced from a bronchiectasis patient.
    • [View Figure] - Figure S2(B) Dot plot from a bronchiectatic patient (day 14) following double staining of induced sputum cells with annexin V (AV) and propidium iodide (PI). Figures indicate percentage of positively stained cells. Quadrant 1=AV-PI+, quadrant 2=AV+PI+, quadrant 3=AV+PI-, quadrant 4=AV-PI-.

Footnotes

  • Funding: Department of Employment and Learning, Northern Ireland Chest, Heart and Stroke Association, Research and Development Office.