Persistent tolerance induced after portal venous injection of allogeneic cells plus cyclophosphamide treatment.

The injection of allogeneic cells via the portal vein (p.v.) is known to reduce responses to donor-alloantigens. In the present study, we have obtained persistent tolerance across Mls and multiple minor histocompatibility complexes by p.v. preimmunization followed by the administration of cyclophosphamide (CY). A hundred percent survival of (BALB/c x DBA/2)F1 (CDF1) skin grafts for more than 200 days was observed when BALB/c mice were preimmunized with spleen cells of CDF1 (3 x 10(7)) via the p.v. and administered 300 mg/Kg CY 2 days after the p.v. injection. Comparable survival of the skin graft was observed when bone marrow cells instead of spleen cells were p.v. preimmunized. However, the survival rate was significantly decreased when LPS-stimulated blastic cells were p.v. preimmunized. Microchimerism has been observed in the liver, thymus, bone marrow and peripheral blood of recipients. V beta 6+ cells decreased in CD4+ cells of recipients of the p.v. preimmunization plus CY treatment. However, there was no difference in the decrease in V beta 6+ cells between recipients accepting the CDF1 skin grafts and recipients that had rejected the skin grafts. Furthermore, no intrathymic depletion of the V beta 6+ cells was observed. From these results, it is suggested that, rather than clonal deletion, other mechanisms such as clonal anergy or suppression may be involved in the induction of persistent tolerance after the p.v. preimmunization plus CY treatment.