Using a novel “Integrated Biomarker Proteomic” index to assess the effects of freshwater pollutants in European eel peripheral blood mononuclear cells

Abstract

Using proteomic data as biomarkers of environmental pollution has the potential to be of a great interest in ecological risk assessment as they constitute early warning indicators of ecologically relevant effects on biological systems. To develop such specific and sensitive biomarkers, the use of a set of proteins is required and the identification of protein expression signatures (PES) may reflect the exposure to specific classes of pollutants. Using 2D-DIGE (Differential in Gel Electrophoresis) methodology, this study aimed at identifying specific PES on European eel (Anguilla anguilla) peripheral blood mononuclear cells (PBMC) after 48h in vitro exposure to two sublethal concentrations of dichlorodiphenyltrichloroethane (p,p′-DDT) (10μg/L and 1mg/L) or cadmium (Cd) (1μg/L and 100μg/L). The present results have been supplemented with data of a first in vitro study on cells exposed to perfluorooctane sulfonate (PFOS) (10μg/L and 1mg/L). A total of thirty-four protein spots, belonging to 18 different identified proteins found in all conditions, have been selected as possible biomarkers to develop a synthetic Integrated Biomarker Proteomic (IBP) index. IBP follows a dose–response relationship with higher values at the highest tested concentration for each pollutant (Cd: 9.96; DDT: 7.44; PFOS: 7.94) compared to the lowest tested concentration (Cd: 3.81; DDT: 2.91; PFOS: 2.06). In a second step, star plot graphs have been applied to proteomic data in order to allow visual integration of a set of early warning responses measured with protein biomarkers. Such star plots permit to discriminate the type of pollutant inducing a proteomic response. We conclude that using IBP is relevant in environmental risk assessment, giving to this index the potential to be applied as a global index of proteome alteration in endangered species such as the European eel. Biological significanceIn this study, 34 protein spots have been selected as possible biomarkers to develop a synthetic Integrated Biomarker Proteomic index (IBP). Results show that IBP follows a dose–response relationship with higher values at the highest tested concentration for each pollutant compared to the lowest tested concentration. Star plot graphs have also been applied to proteomic data in order to allow visual integration of a set of early warning responses measured with protein biomarkers. Such star plots permit to discriminate the type of pollutant inducing a proteomic response. IBP is relevant in environmental risk assessment, giving to this index the potential to be applied as a global index of proteome alteration in endangered species such as the European eel.