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Enterovirus 71 (EV71) is one of the 69 enteroviruses belonging to the picornavirus family. We have previously generated a recombinant baculovirus encoding P1 and 3CD genes for co-expressing P1 and 3CD proteins for VLP assembly in insect cells. In this study, we further characterized the VLP after ultracentrifugation purification, and evaluated the immunological properties of VLPs in Balb/c mice. The immunization experiments show that the VLP induced stronger anti-VP1 antibody titer and neutralization titers compared to the inactivated EV71 virus and lysed VLP. The antibodies induced by EV71 VLP were also capable of neutralizing live EV71 of different strains. Moreover, the VLP was able to induce cellular immunities as judged by helper T cell proliferation assay and cytokine assay. These data collectively confirmed that EV71 VLP produced from baculovirus/insect cells is able to elicit humoral and cellular immunities in mice, implicating its potentials as a vaccine against EV71 infection.