骨髓移植诱导大白兔特异性免疫耐受模型的建立

目的通过供体与受体的骨髓移植来诱导受体对供体的特异性免疫耐受。方法30只雄性和30只雌性新西兰大白兔分别作为供者和受者,随机配对,将30对大白兔再随机分为5组,即无处理对照组、单纯照射组、单纯骨髓移植(BMT)组、实验组(照射+BMT)和无关皮肤供体对照组,每组6对。受者给予7Gy的全身照射(TB I),24小时后耳缘静脉输注骨髓单个核细胞,5天后再次回输相同供者的骨髓单个核细胞,两次总的细胞数量为l~1.5×108/kg,在第二次回输细胞的同时,移植皮肤。观察皮肤移植存活时间、对第三方皮肤的排斥反应及对供体的单向混合淋巴细胞反应,并用多聚酶链式反应(PCR)检测嵌合体的形成。结果单纯照射和骨髓移植不能延长移植物的存活时间,也没有形成供受者混合嵌合;实验组皮肤移植物存活时间(45.83±3.97 d)比无处理对照组(10.83±0.75)、单纯照射组(12.5±1.0488)、单纯骨髓移植组(11.33±0.8165)、无关皮肤供体对照组(12.17±1.169)明显延长(P<0.01)。结论对大白兔模型,采用全身照射加大剂量骨髓移植的方法,可形成较长时间的混合嵌合,并获得一定程度的特异性免疫耐受。

Establishment of specific immunological tolerance model for rabbit by bone marrow transplantation

Objective To investigate the feasibility and practicability of establishing model of specific immunological tolerance using total body irradiation(TBI) and bone marrow transplantation(BUT) for rabbits.Metffods Thirty male and thirty female New Zealand rabbits were respectively used as donors and recipients,matched pairs to them randomly.Thirty pair rabbits were divided randomly into 5 groups,including blank control group,simple TBI group,simple BMT group,experimental group(TBI+BMT) and third party donor control group,with 6 pair of rabbits in each group.Before the BMT,TBI(7Gy)was performed in recipient rabbits.24 hours after TBI,the bone marrow cells were injected through the oto-edge vein.On posttransplant day 5,the bone marrow cells from the same donor were injected again,meanwhile,skin grafting was performed.The total number that bone marrow cells were injected was 1~1.5 × 10~8/kg.The survival time of skin grafts,reject reaction against the third party skin,and mixed lymphocyte reaction(MLR) were observed.The polymerase chain reaction(PCR) was employed for the detection of chimerism.Result Simple BMT or TBI led to neither prolonged survival time of grates nor the formation of chimerism.The skin graft survival time was significantly prolonged in the experimental group(45.83±3.97 d),as compared with the blank control group(10.83 ± 0.75),the TBI group(12.5±1.0488),the BMT group(11.33±0.8165) and the third party donor control group(12.17±1.169),respectively((P<0.01).) Conclusions Our experimental observations suggest that BMT with nonmyeloablative precondition method may induce certain extent of immunological tolerance and prolong the survival time of rabbit skin graft.