Fumonisin B Induces Immunotoxicity and Apoptosis of Chicken Splenic Lymphocytes.

Fumonisin B (FB), produced by , is among the most abundant and toxic mycotoxin contaminations in feed, causing damages to the health of livestock. However, the mechanisms of FB toxicity in chickens are less understood. As splenic lymphocytes play important roles in the immune system, the aim of this study was to investigate the immunotoxic effects and mechanisms of FB on chicken splenic lymphocytes. In the present study, the chicken primary splenic lymphocytes were harvested and treated with 0, 2.5, 5, 10, 20 and 40 μg/mL FB. Then, the cell proliferation, damage, ultrastructure, inflammation and apoptosis were evaluated. Results showed that the proliferation rate of splenic lymphocytes was decreased by FB treatments. The activity of lactate dehydrogenase (LDH) was increased by FB treatments in a dose-dependent manner, implying the induction of cell damage. Consistently, the ultrastructure of splenic lymphocytes showed that FB at all the tested concentrations caused cell structure alterations, including nuclear vacuolation, mitochondrial swelling and mitochondrial crest fracture. Besides, immunosuppressive effects of FB were observed by the decreased concentrations of interleukin-2 (IL-2), IL-4, IL-12 and interferon-γ (IFN-γ) in the cell culture supernatant. Furthermore, apoptosis was observed in FB-treated cells by flow cytometry. The mRNA expressions of apoptosis-related genes showed that the expression of was decreased, while the expressions of the , and were increased with FB treatment. Similar results were found in the concentrations of apoptosis-related proteins in the cell supernatant by ELISA assay. Moreover, regression analysis indicated that increasing FB concentration increased LDH activity, concentrations of Bax, Bak-1 and mRNA expression of linearly, increased M1 area percentage quadratically, decreased concentration of IFN-γ, mRNA expression of linearly, and decreased concentrations of IL-2 and IL-4 quadratically. Besides, regression analysis also showed reciprocal relationships between IL-12 concentration, mRNA expression and increasing FB concentration. The increasing FB concentration could decrease IL-12 concentration and increase mRNA expression. Altogether, this study reported that FB induced the immunotoxicity of chicken splenic lymphocytes and caused splenic lymphocytes apoptosis by the Bcl-2 family-mediated mitochondrial pathway of caspase activation.