Abstract: Objective To establish a mouse model of CD8⁺T cell-mediated acute graft versus host disease（GVHD）. Methods Female C3H.SW（H-2D^b, CD45.2⁺）mice was used as donors and B6/SJL（H-2D^b, CD45.1⁺）mice as recipients, both of them are MHC-identical but miHA-mismatched. Before transplantation, the B6 recipients received ¹³⁷γ ray （9.5 Gy） TBI administered in three treatments from a ¹³⁷Cs source, then the T cell-depleted bone marrow （T^-BM） cells and CD8⁺T cells from C3H.SW donors were injected immediately via tail vein. Results Two of the recipients injected with C3H.SW TBM and C3H.SW CD8⁺T cells died 28 days and one died 70 days after transplantation. The recipients injected only with C3H.SW T^-BM were still alive within 70 days after transplantation. B6 mice without transplantation died within 6-10 days after irradiation. Flow cytometric analysis showed that mononuclear cells from the bone marrow, spleens and livers of B6 mice derived from the donor, and part of mononuclear cells from the spleens and livers was CD45.2⁺，CD8⁺T cell. Pathologic examination of the liver showed that the accumulation of mononuclear cells was in the portal areas of the liver. The structure of skin was destroyed. Conclusions The miHA-mismatched mouse model was CD8⁺T cell-mediated acute GVHD model. The MHC-identical, multiple miHA mismatched mouse model is analogous to human clinical miHA-mismatched bone marrow transplantation, which can be used as a good tool for study of pathogenesis and prophylaxis of GVHD.