Influence of cell dose and graft-versus-host reactivity on rejection rates after allogeneic bone marrow transplantation

Abstract

Abstract The number of cells transplanted and their capacity to induce graft- versus-host reactivity (GvHR) are two factors that are suspected to influence the engraftment of allogeneic bone marrow. We have investigated their impact on graft rejection rates in busulfan-treated LEW rats. In a series of experiments, we varied (1) the number of marrow cells transferred (1, 5, 10, 20, 30, and 40 x 10(7)), (2) the degree of pretransplant immunosuppression (1.5, 3.0, and 4.5 Gy of total body irradiation [TBI]; 0, 30, 60, 90, 120, and 180 mg/kg cyclophosphamide), and (3) the ability of the marrow graft to induce classical GvHR against major histocompatibility complex (MHC) antigens [semiallogeneic (CAP x LEW)F1 or CAP rats as marrow donors]. Reducing either the immunosuppressive pretreatment or the number of cells transplanted resulted in a stepwise increase in rejection rates. However, every reduction in the size of the marrow inoculum was compensated by increased immunosuppression and vice versa. While 60 mg/kg cyclophosphamide was sufficient to prevent rejections after grafting of 40 x 10(7) cells, 90 mg/kg was necessary to ensure 100% engraftment after transplantation of 20 x 10(7) cells, 120 mg/kg after 10 x 10(7) cells, and 180 mg/kg after 1 x 10(7) cells. Since CAP marrow leads to GvHR-mediated immunosuppression in LEW recipients, in contrast to (CAP x LEW)F1 marrow, we had supposed that lower cell numbers or cyclophosphamide doses are sufficient to achieve engraftment of CAP marrow. Although severe GvHR was present in all animals receiving escalating doses of CAP cells, the rejection rates were the same as for (CAP x LEW)F1 marrow. In conclusion, we have demonstrated that there is a sensitive balance between the immunosuppression of the host and the number of marrow cells transferred. We were not able to number of marrow cells transferred. We were not able to detect a beneficial effect of classical GvHR against MHC antigens on the engraftment of allogeneic marrow. Thus, our results do not support the hypothesis that the loss of GvHR-mediated immunosuppression is responsible for higher rejection rates following transplantation of T-cell-depleted bone marrow.