Abstract: Objective To explore whether vimentin (VIM) mediates the activation of inflammasome in mice with EV71 infection in the central nervous system. Methods Forty VIM knockout mice (VIM-/-, 3 to 5 days old) were randomly divided into control group and infection group. The infection group was intraperitoneally injected with EV71 (108 TCID50), while the control group was injected with PBS (10 μL); another 40 wild-type mice (WT, 3 to 5 days old) were grouped in the same manner. The general conditions of mice were observed each day. Western blotting, ELISA, and RT-PCR were used to measure the levels of IL-1β and casepase-1 in the brain or cerebrospinal fluid. The pathological changes in the cerebella and brain were observed using immunohistochemistry. Results Compared with the control group, the VIM-/- mice infected with EV71 showed no significant changes in NLRP3, IL-1β or caspase-1 expression. The WT mice infected with EV71 showed obviously increased NLRP3, IL-1β, and caspase-1 expressions in the central nervous system. The neurons of infected VIM-/- mice exhibited milder cell damage than the those in WT mice. Conclusion VIM mediates the activation of inflammasome and promotes brain inflammation and neuronal damage in mice with EV71 infection in the central nervous system.