Activation of Human Peripheral Blood Eosinophils by Cytokines in a Comparative Time-Course Proteomic/Phosphoproteomic Study

Activated eosinophilscontribute to airway dysfunction and tissue remodelingin asthma and thus are considered to be important factors in asthma pathology. We report here comparative proteomic and phosphoproteomicchanges upon activation of eosinophilsusing eight cytokinesindividually and in selected cytokinecombinations in time-course reactions. Differential protein and phosphoproteinexpressions were determined by mass spectrometry after 2-dimensional gel electrophoresis (2DGE) and by LC–MS/MS. We found that each cytokine-stimulation produced significantly different changes in the eosinophilproteome and phosphoproteome, with phosphoproteomicchanges being more pronounced and having an earlier onset. Furthermore, we observed that IL-5, GM-CSF, and IL-3 showed the greatest change in protein expression and phosphorylation, and this expression differed markedly from those of the other five cytokinesevaluated. Comprehensive univariate and multivariate statisticalanalyses were employed to evaluate the c...