An analysis of the phosphoproteome of immune cell lines exposed to the immunomodulatory mycotoxin deoxynivalenol

Abstract

a b s t r a c t a r t i c l e i n f oThe mycotoxin deoxynivalenol (DON) commonly contaminates cereal grains. It is ubiquitous in the Western European diet, although chronic, low-dose effects in humans are not well described, but immunotoxicity has been reported. In this study, two-dimensional gel electrophoresis was used to identify phosphoproteomic changes in human B (RPMI1788) and T (Jurkat E6.1) lymphocyte cell lines after exposure to modest concentrations of DON (up to 500 ng/mL) for 24 h. Proteins identified as having altered phosphorylation state post-treatment (C-1-tetrahydrofolate synthase, eukaryotic elongation factor 2, nucleoside diphosphate kinase A, heat shock cognate 71 kDa protein, eukaryotic translation initiation factor 3 subunit I and growth factor receptor-bound protein 2) are involved in regulation of metabolic pathways, protein biosynthesis and signaling transduction. All exhibited a greater than 1.4-fold change, reproducible in three separate experiments consisting of 36 gels in total. Flow cytometry validated the observations for eukaryotic elongation factor 2 and growth factor receptor-bound protein 2. These findings provide further insights as to how low dose exposure to DON may affect human immune function and may have potential as mechanismbased phosphoprotein biomarkers for DON exposure.

Global protein phosphorylation dynamics during deoxynivalenol-induced ribotoxic stress response in the macrophage

“…Proteomics facilitates large-scale identification and quantification of proteins, providing information on protein expression and post-translational modification (Farley and Link, 2009; Mallick and Kuster, 2010). Proteome and phosphoproteome changes occurring after prolonged (6 h or 24 h) DON treatment have been previously measured in human B and T cell lines (Osman et al , 2010; Nogueira da Costa et al , 2011a; Nogueira da Costa et al , 2011b). While these studies are important for identification of biomarkers of effect, they are not informative from perspective of early events and signaling within the macrophage, a primary target of DON which mediates innate immune activation (Zhou et al , 2003; Pestka, 2008).…”

The importance of accounting for sex in the search of proteomic signatures of mycotoxin exposure

“…Proteomic analysis also revealed some metabolic changes induced by DON related with an alteration of nucleotide biosynthesis [126]. The same authors later employed the same experimental setup, this time paying more attention to changes in the phosphoproteome [127]. presence of ribotoxins, such as the phosphorylation of key proteins involved in the early inhibition of translation and the recruitment/maintenance of stress-related proteins [44].…”

Proteomics in food: Quality, safety, microbes, and allergens

“…In the last 10 years proteomics has been used either to build a knowledgebase [81] or investigate how different environmental or artificial conditions during fungal growth can modulate fungi stress response [82][83][84] or proteome changes during mycotoxin producing conditions [85][86][87][88]. Recently, several proteomic-based studies have been made to investigate how mycotoxins can modulate immunity in vitro [89,90], mitochondrial dynamics [91], or induce selective toxicity [92][93][94][95].…”