Document Detail

Effects of homogenization of induced sputum by dithiothreitol on polymorphonuclear cells.
MedLine Citation:
PMID:  16204788     Owner:  NLM     Status:  MEDLINE    
Induced sputum represents a useful and non-invasive tool to isolate different cells from the airways. Complete homogenization of sputum is important for dispersion of cells and is usually achieved by use of dithiothreitol (DTT). However, it is not known if DTT will influence the viability and functionality of cells obtained by induced sputum. In the present study, induced sputum was processed by DTT or by PBS treatment. The obtained neutrophils were compared with neutrophils obtained from peripheral blood and from bronchoalveolar lavage fluid (BAL). These isolated neutrophils were treated in a similar way as the sputum neutrophils with DTT or PBS. All isolated cells were used for chemiluminescence tests and for the measurement of elastase and myeloperoxidase release after stimulation with fMLP. The results showed that the maximum chemiluminescence response was always significantly lower after DTT treatment: blood, 16.68 +/-1.89 vs. 2.62 +/-0.43 mV, P<0.0001; sputum, 2.96 +/-0.30 vs. 1.09 +/-0.01 mV, P<0.01; BAL, 25.47 +/-0.88 vs. 8.22+/-0.20 mV, P<0.0001. Both spontaneous and fMLP-induced release of elastase and myeloperoxidase (MPO) was in most cases enhanced after DTT-treatment (P-values range from 0.24 to <0.01). We conclude that the use of DTT to homogenize sputum for dispersion of cells is harmful to cell functions and these cells are hampered for the evaluation of their normal functional characteristics.
F J van Overveld; U Demkow; D Górecka; E Skopinska-Rozewska; W A de Backer; J Zielinski
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of physiology and pharmacology : an official journal of the Polish Physiological Society     Volume:  56 Suppl 4     ISSN:  1899-1505     ISO Abbreviation:  J. Physiol. Pharmacol.     Publication Date:  2005 Sep 
Date Detail:
Created Date:  2005-10-05     Completed Date:  2007-12-06     Revised Date:  2008-05-19    
Medline Journal Info:
Nlm Unique ID:  9114501     Medline TA:  J Physiol Pharmacol     Country:  Poland    
Other Details:
Languages:  eng     Pagination:  143-54     Citation Subset:  IM    
Department of Respiratory Medicine, Antwerp University, Antwerp, Belgium.
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MeSH Terms
Bronchoalveolar Lavage Fluid / cytology*
Cell Separation / methods*
Cell Survival / drug effects
Dithiothreitol / pharmacology*
Expectorants / pharmacology*
Leukocyte Elastase / metabolism
N-Formylmethionine Leucyl-Phenylalanine / pharmacology
Neutrophils / drug effects*,  enzymology
Peroxidase / metabolism
Reactive Oxygen Species / metabolism
Sputum / cytology*
Time Factors
Reg. No./Substance:
0/Expectorants; 0/Reactive Oxygen Species; 3483-12-3/Dithiothreitol; 59880-97-6/N-Formylmethionine Leucyl-Phenylalanine; EC; EC Elastase

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