Proteomic analysis of the effects of the immunomodulatory mycotoxin deoxynivalenol

Abstract

The mycotoxin deoxynivalenol (DON) contaminates cereals worldwide and is a common contaminant in the Western European diet. At high doses, DON induces acute gastrointestinal toxicity; chronic, low-dose effects in humans are not well described, but immunotoxicity has been reported. In this study, 2-DE was used to identify proteomic changes in human B (RPMI1788) and T (JurkatE6.1) lymphocyte cell lines after exposure to minimally toxic concentrations (up to 500 ng/mL) for 24 h. Proteins which changed their abundance post treatment, by a greater than 1.4-fold change reproducible in three separate experiments consisting of 36 gels in total, are ubiquitin carboxyl-terminal hydrolase isozyme L3, proteasome subunit β type-4 and α type-6, inosine-5'-monophosphate dehydrogenase 2, GMP synthase, microtubule-associated protein RP/EB family member 1 (EB1), RNA polymerases I, II, III subunit ABC1, triosephosphate isomerase and transketolase. Flow cytometry was used to validate changes to protein expression, except for EB1. These findings provide insights as to how low-dose exposure to DON may affect human immune function and may provide mechanism-based biomarkers for DON exposure.

GC-TOF/MS-based metabolomics approach to study the cellular immunotoxicity of deoxynivalenol on murine macrophage ANA-1 cells

“…The metabolism results of DON-immunotoxicity were in unexpected accordance with the conclusion of proteomics [25] and genomics [26].…”

“…Collectively, these studies have revealed that the metabolic pathways of oxidative metabolism [20], glycolysis [21], and glutaminolysis [22] preferentially fuel the cell fate decisions and effector functions of immune cells [23]. can produce the impairment of glycolysis (and gluconeogenesis) and, subsequently, lead to reduced energy generation [25]. A research report [26] of gene pathway analysis M A N U S C R I P T…”

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

“…Reduced cell viability due to DON exposure in the current series of experiments was similar to previous data and thus we selected doses and an exposure period (24 h) associated with modest (~20%) reduction in cell viability in the two cell lines representing doses at the lower end of a range used by others [23,24]. The above mentioned exposure period was also chosen based on previous studies analyzing the effects of DON on immune cell lines [16], on reproducibility and reduced variability of cell viability and a significant decrease in cell viability after a 30 h exposure period. As our objective was to induce low to moderate changes to cell viability, a 24 h exposure period was shown to be a reasonable compromise between establishing a real effect consistent with human exposure and the induction of cell death.…”

“…The development and application of "omics" techniques to develop potential mechanism-based biomarkers is a promising approach and we recently identified a number of proteins altered in expression following DON exposure in human cells lines [16]. We now extend this work to compare phosphoprotein expression in human B (RPMI1788) and T (Jurkat E6.1) lymphocyte cell lines exposed to DON at a maximum concentration of 500 ng DON/mL for a period of up to 54 h. These cell types represent major cellular components of the adaptive immune response and have been used extensively previously [17,18].…”

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).…”