| Objective: Study the possibility of selective depletion of alloreactive donor T cells based on expression of two activation-induced antigens (cd25 and cd69) from C57BL/6H-2b mice to prevent graft-versus-host disease (GVHD). Methods: T cells from C57BL/6H-2b were first activated with BALB/c H-2d host spleen cells in a 2-day mixed-lymphocyte-culture (MLC). Following this activation, this compound is selectively depleted based on expression of two activation-induced antigens cd25 and cd69 using magnetic cell sorting. The depleted cells or the untreated cells were then rechallenged respectively in a secondary mixed lymphocyte culture, with the same stimulator cells or a third-party-(DBA H-2k)or tumor-specific(SP2/0, BALB/c-origin myeloma) cells. Cells prolification were assayed at the indicated time points(1,2,3,4,5 days).These treated cells or control-cultured cells (2.0*106) mixed with bone marrow cells(5.0*106) from C57BL/6 were transfused respectively by the trail vain into the lethally irradiated BALB/c mice to observe the survival time, GVHD incidence and pathological analysis. Results: Mixed lymphocyte culture assays demonstrated that this technique led to a significant decrease in alloreactivity of donor cells(29.02～64.17%),which at the same time preserved reactivity against third party cells(49.61～75.69%)and anti-tumor cells(61.14～68.62%).The mice in the group of control-coclutured were died of acute GVHD within 24days. The 7 recipient mice in the treated group were free of acute GVHD, and 3 mice were died of acute GVHD(aGVHD) within 23 days. Conclusions: MACS-based ex-vivo depletion of alloreactive donor T cells based on expression of two activation-induced antigens (cd25 and cd69) could inhibite anti-host responses, by contrast,anti-SP2/O and anti-third-party responses were preserved. Cotransplantation of these selected depleted cells with bone marrow cells could reduce aGVHD.
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