Zearalenone (ZEN), a mycotoxin produced by several Fusarium spp., is most commonly found as a contaminant in stored grain and has chronic estrogenic effects on mammals. In this in vitro study, we compared the effects of zearalenone (ZEN) and some of its derivatives: α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), and zearalanone (ZAN) on several peripheral blood mononuclear cell (PBMC) parameters: cytotoxicity, proliferation, as well as antibody and cytokine synthesis. The amounts of toxins necessary to inhibit viability, in a dehydrogenase enzyme activity assay (MTT test), by 50% were: 22.7 μM for ZEN, 29.1 μM for α-ZOL, 17.3 μM for β-ZOL and 26.3 μM for ZAN. The administration of 10 μM toxin induced a decrease in the ConA stimulated proliferation of PBMC by 19.6% for ZAN, 45.4% for ZEN, 43.6% for α-ZOL and 85.2% for β-ZOL, when compared to the control stimulated cells. Also, ZEN and its metabolites at concentrations higher than 5 μM induced a significant decrease of the IgG, IgA or IgM levels. Concentrations of 5 and 10 μM of ZEN and ZAN significantly decreased the TNF-α synthesis in the supernatant of the stimulated cells; 10 μM of ZAN also decreased IL-8 synthesis. In conclusion, our results show that ZEN and ZEN derivatives altered several parameters of the humoral and cellular immune response. Therefore, our results are clinically relevant as ZEN and its metabolites are frequent contaminants of animal feed and we have shown that intoxicated animals are incapable of inducing an adequate immune response.