Castellani, Stefano

Conese, Massimo

Di Gioia, Sante

Lepore, Silvia

Maffione, Angela Bruna

Martínez, Maria Carmen

Porro, Chiara

Ratclif, Luigi

Trotta, Teresa

Firenze University Press: E-Journals

IJAE Vol. 115, n. 1/2 (Supplement), 2010
142
A new biological marker isolated and characterizated 
in sputum from lung pathology patients: 
microparticles
Chiara Porro1, Silvia Lepore1, Teresa Trotta1, Stefano Castellani1, Luigi Ratclif2, Sante Di Gioia1, Maria 
Carmen Martínez3, Massimo Conese1, Angela Bruna Maffione1
1 Department of Biomedical Sciences, University of Foggia, Foggia, Italy 
2 Centro Regionale di Supporto FC, Ospedale “G. Tatarella”, Cerignola, Italy 
3 CNRS UMR 6214, INSERM U 771, Université d’Angers, Angers, France
Sputum is recognized as a very useful sampling method in Cystic Fibrosis (CF) for 
both research and clinical use aiding both the diagnosis and monitoring of lung disease 
inflammatory status. A great advantage of the technique is that it enables sampling of 
the airways in a non-invasive manner. We investigated on the presence and phenotype 
of microparticles (MPs) in the sputum, of acute and intermittent CF and Primary Ciliary 
Dyskinesia (PCD) adult patients. MPs are membrane vesicles that are released during 
cell activation and apoptosis. MPs have different biological effects depending on the cell 
from they originate. CF lung disease is characterized by massive neutrophil influx in 
the airways, their activation and eventually apoptosis. Spontaneous sputum, obtained 
from 21 CF patients (10 acute and 11 intermittent) and 7 patients with PCD, was lique-
fied with Sputasol. MPs were counted, visualized by electron microscopy, identified 
in the supernatants of treated sputum by cytofluorimetry and immunodecoration for 
leukocyte (CD11a), granulocyte (CD66b), and monocyte-macrophage (CD11b) antigens. 
Electron microscopy revealed that sputum MPs were in the 100-500 nm range and did 
not contain bacteria. CF sputa contained higher number of MPs in comparison with 
PCD sputa. CD66b and CD11a, but not CD11b, levels in MPs were significantly higher 
in CF than in PCD. Acute and intermittent patients presented significantly higher levels 
of CD11a- and CD66b-expressing MPs respect to PCD patients. MPs are detectable in 
sputa obtained from CF patients and are predominantly of granulocyte origin. These 
findings are consistent with massive influx of neutrophils into CF airways and their 
accumulation on the surface of the airway epithelium. In this environment, neutrophils 
are activated by bacterial products, pro-inflammatory cytokines, and chemokines. Neu-
trophils undergo apoptosis, as normally happens in acute inflammation, but also post-
apoptotic necrosis, releasing toxic enzymes and oxygen radicals. MPs in CF sputa likely 
reflect both activation and apoptosis of neutrophils. The significantly less presence of 
granulocyte-derived MPs in PCD respect to CF patients, may be due to the fact that lung 
disease in PCD, although similar, is delayed in time as compared with CF.
In CF airways, neutrophil-derived MPs could contribute to self-perpetuating inflam-
matory cycle, and may account for the exaggerated pro-inflammatory response of cells 
in CF patients. The MPs isolated from sputum could be a new non invasive methods and 
opens a new opportunity for the study of lung disease.
Key words
Microparticles, sputum, cytofluorimetry, electron microscopy


A new biological marker isolated and characterizated in sputum from lung pathology patients: microparticles

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A new biological marker isolated and characterizated in sputum from lung pathology patients: microparticles

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