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Tissue factor (TF) plays an important role in the host's immune system as the principal initiator of coagulation. However, the precise function of TF in teleosts remains unclear. We determined the cDNA sequence of TF from ayu Plecoglossus altivelis (PaTF). The PaTF transcript was expressed in all tested tissues, and changes in expression were observed in tissues and monocytes/macrophages (MO/MΦ) upon infection with Vibrio anguillarum. PaTF was prokaryotically expressed and purified to prepare anti-PaTF antibodies. Western blot analysis revealed that native PaTF was glycosylated in thrombocytes, but not in ayu MO/MΦ. Microparticles could transfer PaTF to thrombocytes. PaTF neutralization or knockdown led to anti-inflammatory status in ayu MO/MΦ upon V. anguillarum infection. PaTF neutralization reduced the apoptosis of ayu MO/MΦ and improve survival rate in V. anguillarum-infected ayu. Our results indicate that PaTF plays a role in ayu immune response against bacterial infection as a pro-inflammatory mediator.