T-2 Toxin Induces Macrophage Extracellular Trap Formation via CLOCK-WNT10B-PAD2/ROS Signaling

T-2 toxin is a highly immunotoxic mycotoxin, yet the mechanisms by which it induces macrophage extracellular trap (MET) formation and the involvement of circadian regulator CLOCK remain unexplored. Using RAW264.7 and THP-1 macrophages, we analyzed the kinetic features and molecular regulation of T-2 toxin (14-32 nM, 24 h)-induced MET formation. We showed that T-2 toxin induced METs in a time-dependent manner with 4 and 8 h as critical windows. The PAD2 pathway continuously drives this process, with ROS playing a limited role. Mechanistically, T-2 toxin suppressed CLOCK expression, relieving its transcriptional repression of WNT10B, which activated downstream PAD2 and ROS pathways to cooperatively promote MET formation. Clock knockdown significantly enhanced MET release, confirming its negative regulatory function. Together, our findings demonstrate that T-2 toxin suppresses CLOCK expression, relieving WNT10B repression and driving MET generation. This work advances our understanding of the mechanism underlying T-2 toxin-induced MET formation.