Macrophage migration inhibitory factor (MIF) has been characterized as a proinflammatory cytokine. Previous studies have indicated that MIF may play a beneficial role or a detrimental role in microbial infections, depending on pathogens. In this study, we investigated the role of MIF in Listeria monocytogenes infection. The MIF titers increased 6h after lethal L. monocytogenes infection but not in the sublethal infection. The elimination of bacteria from the spleens and livers was not affected by anti-MIF antibody (Ab) injection in the sublethal infection, whereas anti-MIF Ab treatment rescued mice from the lethal infection, suggesting that MIF plays a deteriorating role in lethal L. monocytogenes infection. Anti-MIF Ab treatment significantly augmented interleukin (IL)-10 production in the spleens and livers 24h after infection, suggesting that MIF might down-regulate IL-10 production. Although the administration of anti-IL-10 monoclonal Ab showed no significant effect on the bacterial growth in the organs, the bacterial infection was deteriorated by the combined administration of Abs against MIF and IL-10. On the other hand, anti-MIF Ab treatment also increased in the serum cortisol titer 6h after infection compared with the control immunoglobulin G-injected group. Depletion of endogenous IL-10 decreased serum cortisol titers. These results suggested that IL-10 and cortisol might be involved in the deteriorating effect of MIF on lethal L. monocytogenes infection.