Isolation and Characterization of Microparticles in Sputum from Cystic Fibrosis Patients

Overview

Journal Respir Res

Specialty Pulmonary Medicine

Date 2010 Jul 13

PMID 20618958

Citations 32

Authors

Chiara Porro

Silvia Lepore

Teresa Trotta

Stefano Castellani

Luigi Ratclif

Anna Battaglino

Sante Di Gioia

Maria C Martinez

Massimo Conese

Angela B Maffione

Affiliations

Soon will be listed here.

Abstract

Background: Microparticles (MPs) are membrane vesicles released during cell activation and apoptosis. MPs have different biological effects depending on the cell from they originate. Cystic fibrosis (CF) lung disease is characterized by massive neutrophil granulocyte influx in the airways, their activation and eventually apoptosis. We investigated on the presence and phenotype of MPs in the sputum, a rich non-invasive source of inflammation biomarkers, of acute and stable CF adult patients.

Methods: Spontaneous sputum, obtained from 21 CF patients (10 acute and 11 stable) and 7 patients with primary ciliary dyskinesia (PCD), was liquefied with Sputasol. MPs were counted, visualized by electron microscopy, and identified in the supernatants of treated sputum by cytofluorimetry and immunolabelling for leukocyte (CD11a), granulocyte (CD66b), and monocyte-macrophage (CD11b) antigens.

Results: Electron microscopy revealed that sputum MPs were in the 100-500 nm range and did not contain bacteria, confirming microbiological tests. CF sputa contained higher number of MPs in comparison with PCD sputa. Levels of CD11a+-and CD66b+-, but not CD11b+-MPs were significantly higher in CF than in PCD, without differences between acute and stable patients.

Conclusions: In summary, MPs are detectable in sputa obtained from CF patients and are predominantly of granulocyte origin. This novel isolation method for MPs from sputum opens a new opportunity for the study of lung pathology in CF.

Citing Articles

Extracellular vesicles in sputum of children with cystic fibrosis pulmonary exacerbations.

Ben-Meir E, Antounians L, Eisha S, Ratjen F, Zani A, Grasemann H ERJ Open Res. 2024; 10(6).

PMID: 39655173 PMC: 11626615. DOI: 10.1183/23120541.00547-2024.

Gene expression profile analysis of severe influenza-based modulation of idiopathic pulmonary fibrosis.

Kong J, Chen L Eur J Med Res. 2024; 29(1):501.

PMID: 39420432 PMC: 11488079. DOI: 10.1186/s40001-024-02107-9.

Airway inflammation accelerates pulmonary exacerbations in cystic fibrosis.

Liou T, Argel N, Asfour F, Brown P, Chatfield B, Cox D iScience. 2024; 27(3):108835.

PMID: 38384849 PMC: 10879674. DOI: 10.1016/j.isci.2024.108835.

Laboratory Tools to Predict CFTR Modulator Therapy Effectiveness and to Monitor Disease Severity in Cystic Fibrosis.

Bacalhau M, Camargo M, Lopes-Pacheco M J Pers Med. 2024; 14(1).

PMID: 38248793 PMC: 10820563. DOI: 10.3390/jpm14010093.

Impact of extracellular vesicles on the pathogenesis, diagnosis, and potential therapy in cardiopulmonary disease.

Soto-Vazquez Y, Genschmer K Front Pharmacol. 2023; 14:1081015.

PMID: 36891265 PMC: 9986338. DOI: 10.3389/fphar.2023.1081015.

References

Khan T, Wagener J, Bost T, Martinez J, Accurso F, Riches D . Early pulmonary inflammation in infants with cystic fibrosis. Am J Respir Crit Care Med. 1995; 151(4):1075-82. DOI: 10.1164/ajrccm/151.4.1075. View

Watt A, Courtney J, Moore J, Ennis M, Elborn J . Neutrophil cell death, activation and bacterial infection in cystic fibrosis. Thorax. 2005; 60(8):659-64. PMC: 1747479. DOI: 10.1136/thx.2004.038240. View

Mostefai H, Andriantsitohaina R, Martinez M . Plasma membrane microparticles in angiogenesis: role in ischemic diseases and in cancer. Physiol Res. 2008; 57(3):311-320. DOI: 10.33549/physiolres.931533. View

Conese M, Copreni E, Di Gioia S, De Rinaldis P, Fumarulo R . Neutrophil recruitment and airway epithelial cell involvement in chronic cystic fibrosis lung disease. J Cyst Fibros. 2004; 2(3):129-35. DOI: 10.1016/S1569-1993(03)00063-8. View

Tual-Chalot S, Guibert C, Muller B, Savineau J, Andriantsitohaina R, Martinez M . Circulating microparticles from pulmonary hypertensive rats induce endothelial dysfunction. Am J Respir Crit Care Med. 2010; 182(2):261-8. DOI: 10.1164/rccm.200909-1347OC. View

Vandivier R, Richens T, Horstmann S, deCathelineau A, Ghosh M, Reynolds S . Dysfunctional cystic fibrosis transmembrane conductance regulator inhibits phagocytosis of apoptotic cells with proinflammatory consequences. Am J Physiol Lung Cell Mol Physiol. 2009; 297(4):L677-86. PMC: 2770781. DOI: 10.1152/ajplung.00030.2009. View

Petit-Bertron A, Tabary O, Corvol H, Jacquot J, Clement A, Cavaillon J . Circulating and airway neutrophils in cystic fibrosis display different TLR expression and responsiveness to interleukin-10. Cytokine. 2007; 41(1):54-60. DOI: 10.1016/j.cyto.2007.10.012. View

Hartl D, Latzin P, Hordijk P, Marcos V, Rudolph C, Woischnik M . Cleavage of CXCR1 on neutrophils disables bacterial killing in cystic fibrosis lung disease. Nat Med. 2007; 13(12):1423-30. DOI: 10.1038/nm1690. View

Levy H, Kalish L, Huntington I, Weller N, Gerard C, Silverman E . Inflammatory markers of lung disease in adult patients with cystic fibrosis. Pediatr Pulmonol. 2007; 42(3):256-62. PMC: 4469989. DOI: 10.1002/ppul.20563. View

10.

Wolter J, Rodwell R, Bowler S, McCormack J . Cytokines and inflammatory mediators do not indicate acute infection in cystic fibrosis. Clin Diagn Lab Immunol. 1999; 6(2):260-5. PMC: 95697. DOI: 10.1128/CDLI.6.2.260-265.1999. View

11.

Livraghi A, Randell S . Cystic fibrosis and other respiratory diseases of impaired mucus clearance. Toxicol Pathol. 2007; 35(1):116-29. DOI: 10.1080/01926230601060025. View

12.

Cohen Z, Gonzales R, Davis-Gorman G, Copeland J, McDonagh P . Thrombin activity and platelet microparticle formation are increased in type 2 diabetic platelets: a potential correlation with caspase activation. Thromb Res. 2002; 107(5):217-21. DOI: 10.1016/s0049-3848(02)00334-1. View

13.

Sagel S . Noninvasive biomarkers of airway inflammation in cystic fibrosis. Curr Opin Pulm Med. 2003; 9(6):516-21. DOI: 10.1097/00063198-200311000-00012. View

14.

Bastarache J, Fremont R, Kropski J, Bossert F, Ware L . Procoagulant alveolar microparticles in the lungs of patients with acute respiratory distress syndrome. Am J Physiol Lung Cell Mol Physiol. 2009; 297(6):L1035-41. PMC: 2793184. DOI: 10.1152/ajplung.00214.2009. View

15.

Bonfield T, Panuska J, Konstan M, Hilliard K, Hilliard J, Ghnaim H . Inflammatory cytokines in cystic fibrosis lungs. Am J Respir Crit Care Med. 1995; 152(6 Pt 1):2111-8. DOI: 10.1164/ajrccm.152.6.8520783. View

16.

Vandivier R, Fadok V, Hoffmann P, Bratton D, Penvari C, Brown K . Elastase-mediated phosphatidylserine receptor cleavage impairs apoptotic cell clearance in cystic fibrosis and bronchiectasis. J Clin Invest. 2002; 109(5):661-70. PMC: 150889. DOI: 10.1172/JCI13572. View

17.

Agouni A, Lagrue-Lak-Hal A, Ducluzeau P, Mostefai H, Draunet-Busson C, Leftheriotis G . Endothelial dysfunction caused by circulating microparticles from patients with metabolic syndrome. Am J Pathol. 2008; 173(4):1210-9. PMC: 2543087. DOI: 10.2353/ajpath.2008.080228. View

18.

Janowska-Wieczorek A, Majka M, Kijowski J, Baj-Krzyworzeka M, Reca R, Turner A . Platelet-derived microparticles bind to hematopoietic stem/progenitor cells and enhance their engraftment. Blood. 2001; 98(10):3143-9. DOI: 10.1182/blood.v98.10.3143. View

19.

Benameur T, Andriantsitohaina R, Martinez M . Therapeutic potential of plasma membrane-derived microparticles. Pharmacol Rep. 2009; 61(1):49-57. DOI: 10.1016/s1734-1140(09)70006-4. View

20.

Mutschler D, Larsson A, Basu S, Nordgren A, Eriksson M . Effects of mechanical ventilation on platelet microparticles in bronchoalveolar lavage fluid. Thromb Res. 2003; 108(4):215-20. DOI: 10.1016/s0049-3848(03)00005-7. View