Spred2 controls the severity of Concanavalin A-induced liver damage by limiting interferon-gamma production by CD4⁺ and CD8⁺ T cells

Introduction: Mitogen-activated protein kinases (MAPKs) are involved in T cell-mediated liver damage. However, the inhibitory mechanism(s) that controls T cell-mediated liver damage remains unknown. Objectives: We investigated whether Spred2 (Sprouty-related, EVH1 domain-containing protein 2) that negatively regulates ERK-MAPK pathway has a biological impact on T cell-mediated liver damage by using a murine model. Methods: We induced hepatotoxicity in genetically engineered mice by intravenously injecting Concanavalin A (Con A) and analyzed the mechanisms using serum chemistry, histology, ELISA, qRT-PCR, Western blotting and flow cytometry. Results: Spred2-deficient mice (Spred2^-/-) developed more sever liver damage than wild-type (WT) mice with increased interferon-γ (IFNγ) production. Hepatic ERK phosphorylation was enhanced in Spred2^-/- mice, and pretreatment of Spred2^-/- mice with the MAPK/ERK inhibitor U0126 markedly inhibited the liver damage and reduced IFNγ production. Neutralization of IFNγ abolished the damage with decreased hepatic Stat1 activation in Spred2^-/- mice. IFNγ was mainly produced from CD4⁺ and CD8⁺ T cells, and their depletion decreased liver damage and IFNγ production. Transplantation of CD4⁺ and/or CD8⁺ T cells from Spred2^-/- mice into RAG1^-/- mice deficient in both T and B cells caused more severe liver damage than those from WT mice. Hepatic expression of T cell attractants, CXCL9 and CXCL10, was augmented in Spred2^-/- mice as compared to WT mice. Conversely, liver damage, IFNγ production and the recruitment of CD4⁺ and CD8⁺ T cells in livers after Con A challenge were lower in Spred2 transgenic mice, and Spred2-overexpressing CD4⁺ and CD8⁺ T cells produced lower levels of IFNγ than WT cells upon stimulation with Con A in vitro. Conclusion: We demonstrated, for the first time, that Spred2 functions as an endogenous regulator of T cell IFNγ production and Spred2-mediated inhibition of ERK-MAPK pathway may be an effective remedy for T cell-dependent liver damage.