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Methodology for Sputum Induction and Laboratory Processing
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Sputum Cytokine Repeatability in COPD.

Alex Mulvanny1, Augusta Beech1,2, Jian Li2

  • 1Bioanalytical Department, Medicines Evaluation Unit, Manchester, UK.

International Journal of Chronic Obstructive Pulmonary Disease
|November 27, 2025
PubMed
Summary

Sputum biomarker measurements in COPD patients showed high repeatability over six months. These reproducible inflammatory profiles in sputum link to clinical factors like neutrophilia and bacterial colonization.

Keywords:
biomarkerinflammationluminexrespiratoryvalidation

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Area of Science:

  • Pulmonary Medicine
  • Biomarker Discovery
  • Chronic Obstructive Pulmonary Disease (COPD)

Background:

  • Sputum biomarker measurements are crucial for assessing airway inflammation in COPD patients.
  • A previously validated Luminex assay quantifies 15 analytes in COPD sputum supernatant, revealing profiles linked to neutrophilia, bacterial colonization, and smoking.

Purpose of the Study:

  • To assess the reproducibility of sputum biomarker measurements over a six-month period in COPD patients.
  • To identify correlations between sputum inflammatory markers and clinical characteristics at six months.

Main Methods:

  • Repeat sputum supernatant analysis was performed on 48 COPD patients after six months.
  • A multiplex immunoassay quantified 15 analytes in the sputum samples.
  • Statistical analysis, including intraclass correlation coefficient (ICC), was used to assess repeatability and correlations.

Main Results:

  • The majority of sputum analytes demonstrated good to excellent repeatability between baseline and six months (ICC).
  • Specific cytokines (IL-1β, IL-2, IL-8, IL-17A, G-CSF, MIP-1α, MIP-1β, TNF-α) correlated with sputum neutrophil percentage at six months.
  • Elevated levels of certain cytokines were observed in patients with *Haemophilus influenzae* colonization and in ex-smoking COPD patients.

Conclusions:

  • Multiplex immunoassay of sputum biomarkers shows high reproducibility in COPD patients over time.
  • Reproducible inflammatory signatures in sputum are associated with key clinical characteristics in COPD, including neutrophilia, bacterial colonization, and smoking status.