Journal
CYTOKINE
Volume 56, Issue 2, Pages 422-429Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.cyto.2011.06.025
Keywords
Neutrophil; 2D-DIGE; TNF alpha; GM-CSF; Proteome profiles
Funding
- Dutch Asthma Foundation [3.2.03.63]
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Increased serum levels of TNF alpha and GM-CSF are found in various chronic inflammatory diseases and these cytokines affect the function of circulating and tissue neutrophils. TNF alpha- and GM-CSF-induced protein expression profiles could, therefore, serve as biomarker for the action of these cytokines in vivo. We stimulated human peripheral neutrophils with TNF alpha and GM-CSF in vitro and analyzed changes in their proteome by fluorescence two-dimensional difference gel electrophoresis (2D-DIGE). We report the differential expression of 3 and 18 protein spots following TNF alpha and GM-CSF stimulation, respectively. Differences in protein expression induced by TNF alpha were limited and did not show discriminatory power in a principal component analysis, whereas the profile induced by GM-CSF did. TNF alpha- and GM-CSF-induced both de novo IL-1 beta and sIL-1Ra protein expression as detected by Western blot analysis, which confirmed proper neutrophil activation by these cytokines in vitro. Mass spectrometry analysis of cytokine-regulated protein spots resulted in the identification of 8 proteins. Among the identified proteins, enolase 1 and annexin A1 might function as markers for peripheral neutrophil activation. In conclusion, a proteomic analysis of neutrophils by 2D-DIGE provides proof-of-principle that cytokine-induced protein profiles can serve as biomarkers for the action of individual cytokines in vivo. (C) 2011 Elsevier Ltd. All rights reserved.
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