TDK conference 2020

T-2 toxin contamination of feed is a major concern worldwide resulting in serious impairments regarding animal health as well as the economy. The harmful effects often manifest in hardly recognizable, non-specific symptoms and presence of the toxin can be a predisposing factor in the development of further, secondary diseases. The aim of our study was to investigate the immunomodulatory and cytotoxic effects of T-2 toxin, using primary mono-layer hepatocyte mono-culture and hepatocyte — nonparenchymal cell co-culture models, originated from broiler chicken. The established cell cultures are suitable models to study the function of liver, playing central role in the metabolism of the toxin, moreover, to study further cellular mechanisms involved in the inflammatory and stress responses.

Cultures were exposed to 10, 100 and 1000 nmol/l T-2 toxin containing cell culture medium for 8 and 24 hours. Alterations of cellular metabolic activity were measured using CCK-8 assay. Production of H2O2 as the major reactive oxygen species (ROS) was detected by Amplex Red method. The concentrations of heat shock protein 70 (HSP70) and different pro-inflammatory cytokines such as interleukin (IL-)6 and IL-8 were determined from culture media by chicken specific ELISA tests.

As the result of T-2 administration, metabolic activity was significantly decreased in all the cell culture models in case of every applied concentration and incubation time. The concentrations of HSP70 and IL-8 were increased in hepatocyte mono-cultures in the higher concentration T-2 toxin treated cultures (100 and 1000 nmol/l for HSP70 and 1000 nmol/l for IL-8) after 24 hours incubation. IL-6 levels were intensely elevated in case of 100 and 1000 nmol/l T-2 toxin administration in both of mono- and co-cultures, but only after 8 hours of incubation time. Despite the general harmful effects of T-2 toxin treatment, no significant alteration was observed in ROS production. Furthermore, the different cell cultures showed significantly varying levels of H2O2, HSP70 and IL-8 regardless the applied toxin supplementation.

In conclusion, the established primary cell cultures proved to be proper models to study the specific molecular effects caused by T-2 toxin. Based on our results, the metabolic activity and immune status of the cells were mainly affected by the toxin. However, no changes were found in the extracellular H2O2 levels, which can suggest that ROS production may not play a key mediatory role in the cytotoxic effects of T-2 toxin in chicken liver. The exact effects of T-2 toxin were successfully monitored on the cellular level, and hopefully, our results can contribute to understand the mycotoxin related health and perfomance problems of broilers.