Background: Cytokines have been implicated in the progression of T cell-mediated liver injury, in which mitogen-activated protein kinase (MAPK) cascades may play a role. We focus on the role of Spred-2, a negative regulator of MAPK that constitutively expressed in the liver, in concanavalin A (ConA)-induced acute liver injury. Methods: Wild type(C57BL/6J) and Spred2-KO mice were intravenously injected with ConA (15mg/kg). For some experiments, CD4+T cells, CD8+T cells and Kupffer cells were deleted prior to Con A challenge. Liver injury was assessed by level of alanine aminotransferase and histology. Cytokines/chemokines were measured by ELISA and real-time PCR. Hepatic lymphocytes were analyzed by flow cytometry. Results: Liver injury after 15mg/kg ConA injection in Spred-2KO mice was much severer than that in the control WT mice, as evidenced by increased serum levels of ALT and exacerbated injured area. Activities of caspase-3, -8, and -9 were significantly elevated in Spred-2KO liver. In Spred-2KO mice, systemic level of IFNg was significantly higher than that in WT mice. FACS analyses demonstrated that the numbers of hepatic CD4+T, CD8+T and NKT cells in Spred-2KO liver were significantly increased as compared to WT control. Interestingly, expressions of IFNg and perforin were augmented in CD8+T cells but not CD4+T cells, isolated from Spred-2 KO liver. Neutralization of CD8+T cells but not CD4+T cells restored ConA-induced hepatotoxity. When Kupffer cells were deleted, levels of IFNg-inducing chemokines CXCL9/10 were decreased relative to the control. Upon stimulation with ConA, Kupffer cells from Spred-2KO mice produced higher levels of CXCL9/10 compared with WT mice. Upon stimulation with IFNg, hepatocytes from Spred-2 KO mice and WT mice produced comparable levels of CXCL9/10. Conclusion: Altogether, these results suggest that lacking Spred-2 is deleterious in ConA-induced liver injury, by attracting and activating CD8+T cells via enhanced production of CXCL9/10 from Kupffer cells.