Induction Of Rat Limb Allotransplantation Tolerance By Donor Specific Spleen Cell Tranfusion

Objective To investigate the effect and mechanisms of transplantation tolerance induced by donor specific spleen cell transfusion(DST)combined with cyclophosphamide(CP).Methods 20 male Wistar rats and 20 female SD rats were respectively used as donors and recipients of limb allotransplantation,and another 2 male DA rats were used as the third germ line to detect the specificity of tolerance.20 recipient rats were divided randomly into 4 groups:blank control group,spleen cell group,cyclophosphamide group and experimental group(SC+CP),with 5 rats in each group.On the transplantation day a bonus of 3×10~8 spleen cell were injection through the tail vein,and intraperitoneal infusion of CP(150mg/kg)on day 2.The survival time of limb transplantation,and mixed lymphocyte reaction (MLR)were observed.The polymerase chain reaction(PCR)was employed for the detection of microchimerism.Results The limb graft survival time was significantly prolonged in the experimental group(18.4±1.82d),as compared with the biank control group(8.4±1.14d),the SC group(9.0±1.23d),and the CP group (9.6±1.14d),respectively(P＜0.01).MLR demonstrated that the donor specific inhibition of lymphocyte proliferation in the experimental group (74.2±4.92%)was significantly higher than that in the SC group (37.2±3.90%)and the CP group(39.4±4.16%),respectively(P＜0.01). Microchimerism developed in the SC group,the CP group,and the experimental group.Conclusion 1 The rejection followed by transplanting the hind limb of Wistar rat- SD rat is intense and constancy.The transplanted limb is easy to observe.Several kinds of serum detection can be made on mice.Thus,the model is the ideal animal model of translanted limb.2 The immune tolerance of recipient rat can be induced by donor specific spleen cell tranfusion with the company of abdominal injection of cyclophosphamide.3 The immune tolerance induced by this experiment is partial,ephemeral and specificity for the donor.4 The comformation of the Microchimerism and the level of the engomphosis may be related to the generation and maintain of immune tolerance.