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Isolation and characterization of microparticles in sputum from cystic fibrosis patients

Chiara Porro1, Silvia Lepore, Teresa Trotta

  • 1Department of Biomedical Sciences, University of Foggia, Foggia, Italy. c.porro@unifg.it

Respiratory Research
|July 13, 2010
PubMed
Abstract

Insights

Microparticles (MPs) are detectable in cystic fibrosis (CF) sputum, primarily originating from granulocytes. This finding offers a new avenue for studying CF lung disease through non-invasive sputum analysis.

Area of Science:

  • Cell Biology
  • Immunology
  • Pulmonology

Background:

  • Microparticles (MPs) are cellular vesicles released during cell activation and apoptosis, with functions varying by origin.
  • Cystic Fibrosis (CF) lung disease involves significant neutrophil influx, activation, and apoptosis in airways.
  • Sputum serves as a valuable, non-invasive source for inflammation biomarker discovery in CF.

Purpose of the Study:

  • To investigate the presence and phenotype of MPs in the sputum of adult CF patients.
  • To compare MP profiles in acute and stable CF disease states.
  • To establish a novel method for MP isolation from sputum for CF research.

Main Methods:

  • Sputum samples from 21 CF patients and 7 primary ciliary dyskinesia (PCD) patients were processed.
  • Microparticles were quantified, visualized via electron microscopy, and identified using flow cytometry and immunolabelling (CD11a, CD66b, CD11b).
  • Bacterial presence in MPs was assessed to rule out contamination.

Main Results:

  • Electron microscopy confirmed sputum MPs ranged from 100-500 nm and were bacteria-free.
  • CF sputa exhibited significantly higher MP counts compared to PCD sputa.
  • Elevated levels of CD11a+ and CD66b+ MPs were observed in CF patients versus PCD patients, irrespective of disease state.

Conclusions:

  • Microparticles are detectable in CF sputum and are mainly of granulocyte origin.
  • The study presents a novel method for isolating MPs from sputum, enabling further research into CF lung pathology.
  • MP analysis in sputum may provide new insights into the inflammatory processes in cystic fibrosis.

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