Impact of the Tobacco-Smoking Status on the Phagocyte-Derived Generation of Reactive Oxygen Species: New Modification of the Whole-Blood Chemiluminescence Assay.

A new modification of the whole-blood chemiluminescence assay has been developed and used to assess the impact of the tobacco-smoking status on the phagocyte-derived generation of reactive oxygen species (ROS). The strategy behind experiments consisted of maximizing the preservation of the whole-blood composition in the experimental samples while removing erythrocytes that distort the results of chemiluminescence analysis. The removal of erythrocytes was achieved through their spontaneous sedimentation within 2-3 h. In this approach, only a nominal loss of leukocytes (1%-2% of the total amount), as a result of their co-precipitation with red blood cells, takes place, which does not significantly affect the outcome of the chemiluminescence assay. Luminol was utilized as the appropriate chemiluminophore to monitor the ROS generation in the experimental blood samples. The developed experimental approach enables establishing the differences in the reactivity (priming) of circulating leukocytes in the blood of former smokers, individuals who never smoked, and active smokers, while using traditional and well-studied (zymosan) phagocytosis inducers and new agonists such as particles of tobacco dust from regular cigarettes.