Preliminary investigations on the single and combined cytotoxic effect of T-2 and HT-2 measured by Methyl Thiazol Tetrazolium (MTT) cytotoxicity test using pig lymphocytes

Fusariotoxins are frequently existing contaminants in cereal and other plant products. High relationship was found between the levels of the trichothecene T-2 and HT-2 toxin detected in contaminated commodities. Many in vitro studies investigated the cytotoxicity of T-2 and HT-2, but the interaction between them has not been thoroughly studied yet. The aim of the study was to examine the dose dependent cytotoxic effect of T-2 and HT-2 toxins, single and in combination, using pig lymphocytes in the methyl thiazol tetrazolium (MTT ) assay. The mycotoxins were added at various concentrations, i.e. 0.5, 0.1, 0.05, 0.01 and 0.001 μM of T-2; 1.0, 0.5, 0.2., 0.1 and 0.05 μM of HT-2; and both mycotoxins combined. Two exposure times (6 and 24 h) were tested. Both T-2 and HT-2 toxins exerted a dose dependent effect. After 6 h incubation, the increase in concentration of T-2 from 0.001 to 0.5 μM and HT-2 for 0.05 to 1.0 μM resulted in lower cell viability by 22 and 17%, respectively. After 24 h cell viability was significantly lower compared to values obtained at 6 h, except 0.5 μM T-2 and 0.05, 1.0 μM HT-2, respectively. Measured cell viability for combinations of T-2 and HT-2 was higher compared to the calculated expected values. The two toxins caused lower cell survival when applied together than in single administration after 6 h incubation. After 24 h incubation this tendency was not consistent.