Impact of chemical mixtures from wastewater treatment plant effluents on human immune cell activation: An effect-based analysis

Background: Humans are exposed to many different chemicals on a daily basis, mostly as chemical mixtures, usually from food, consumer products and the environment. Wastewater treatment plant effluent contains mixtures of chemicals that have been discarded or excreted by humans and not removed by water treatment. These effluents contribute directly to water pollution, they are used in agriculture and may affect human health. The possible effect of such chemical mixtures on the immune system has not been characterized. Objective: The aim of this study was to investigate the effect of extracts obtained from four European wastewater treatment plant effluents on human primary immune cell activation. Methods: Immune cells were exposed to the effluent extracts and modulation of cell activation was performed by multi-parameter flow cytometry. Messenger-RNA (mRNA) expression of genes related to immune system and hormone receptors was measured by RT-PCR. Results: The exposure of immune cells to these extracts, containing 339 detected chemicals, significantly reduced the activation of human lymphocytes, mainly affecting T helper and mucosal-associated invariant T cells. In addition, basophil activation was also altered upon mixture exposure. Concerning mRNA expression, we observed that 12 transcripts were down-regulated by at least one extract while 11 were up-regulated. Correlation analyses between the analyzed immune parameters and the concentration of chemicals in the WWTP extracts, highlighted the most immunomodulatory chemicals. Discussion: Our results suggest that the mixture of chemicals present in the effluents of wastewater treatment plants could be considered as immunosuppressive, due to their ability to interfere with the activation of immune cells, a process of utmost importance for the functionality of the immune system. The combined approach of immune effect-based analysis and chemical content analysis used in our study provides a useful tool for investigating the effect of environmental mixtures on the human immune response.