A new rapid staining and measuring method has been developed for the quantification of migrated cells in a microchemotaxis chamber. The migrated cells were, after staining, evaluated by a transmission densitometer. The method introduced here is more accurate and faster than those described previously. In addition the technique can be used to determine the adherent capacity of cells.
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Several chemoattractants have been measured in exhaled breath condensate (EBC) from
patients with COPD. The aim of this study was to compare the eosinophil and neutrophil chemotactic activity contained in EBC from healthy subjects and patients with COPD.
Methods: EBC collected using a commercially available condenser (EcoScreen; Erich Jaeger Viasys; Hoechberg, Germany) was compared in 45 COPD patients and 65 healthy subjects. EBC chemotactic activity for eosinophils and neutrophils was assessed using microchambers (Boyden; Neuro Probe; Gaithersburg, MD). Chemotactic index (CI) was used to evaluate cell migration.
Results: EBC from patients with COPD (CI, 2.210.16 [meanSEM]) and healthy subjects (CI, 1.670.11) displayed significant neutrophil chemotactic activity (p<0.0001 for both), which was however higher in patients with COPD (p<0.001). Healthy smokers had a significantly raised CI for neutrophils by comparison with healthy nonsmokers (p<0.01) and ex-smokers (p<0.05). Likewise, current COPD smokers tended to have greater neutrophil CI than COPD who stopped smoking (p0.08). COPD ex-smokers had raised chemotactic activity by comparison with healthy ex-smokers (p<0.05). Anti–interleukin-8 (106 g/mL) antibodies reduced neutrophil chemotactic activity by 35.2% (p<0.05). EBC also contained significant eosinophil chemotactic activity in healthy subjects (CI, 1.680.09; p<0.0001) and patients with COPD (CI, 1.230.07; p<0.01), with a significantly lower CI in patients with COPD as compared to healthy subjects (p<0.001). Smoking did not influence eosinophil
chemotactic activity in healthy subjects or patients with COPD.