犬肾移植致敏模型的建立

Establishment of Sensitized Canine Model for Kidney Transplantation

目的建立犬致敏后的肾移植动物模型。方法雄性家犬各6条配对作为淋巴细胞供受体,采用小剂量（0.4×10^7-1.2×10^7个/kg）淋巴细胞多部位多次输注的方法诱导致敏。用补体依赖的淋巴细胞毒（complement deendent cytotoxicity,CDC）和混合淋巴细胞培养（mixed lymphocyte culture,MLC）试验进行检测。当CDC转为阳性和MLC显示反应淋巴细胞增殖明显活跃后,淋巴细胞供受体犬间进行交叉肾移植,术后定期ECT动态显像观察移植肾功能变化;分批摘取移植肾行病理检查,观察排斥反应发生情况。结果实验犬均在输注淋巴细胞3-4次后CDC转为阳性,MLC显示反应淋巴细胞增殖明显活跃。肾移植术后4天ECT检查有4条致敏犬移植肾血流灌注下降,肾功能出现损害,摘取的3只移植肾病理检查表现为抗体介导的排斥（延缓）或急性排斥反应;对照组未见异常。术后7天,致敏犬移植肾均因急性排斥失功;对照组3条犬有2条出现肾功能损害,病理检查示急性排斥反应。结论小剂量淋巴细胞多部位多次输注可以诱导出家犬的免疫致敏状态。致敏犬肾移植术后移植肾排斥反应出现较早,肾功能损害更严重。

Objective To establish a sensitized canine model for kidney transplantation.Methods 12 male dogs were averagely grouped as donors and recipients.A small number of lymphocytes of donor canine（0.4×10^7-1.2×10^7/kg）were injected into different anatomic of the paired recipient canine weekly.Specific immune sensitization was monitored by complement dependent cytotoxicity（CDC）and mixed lymphocyte culture（MLC）test.When results of CDC test converted positive and MLC test showed a significant proliferation of reactive lymphocytes of canine recipients,the right kidneys of the paired dogs were excised and transplanted to each other concurrently.Injury of renal allograft function was determined by ECT dynamic kidney photography and pathologic investigation.Results After 3 to 4 times of canine donor lymphocyte stimulations,CDC results usually converted positive and reactive lymphocytes of canine recipients were also observed to proliferate significantly by MLC test.Renal allograft function was observed to be deteriorated 4 days post-operatively in 4 of 6 canine recipients,and no deterioration was observed in control dogs.Pathologic changes suggested antibody-mediated rejection（delayed）or acute rejection in the excised renal allografts of 3 sensitized dogs.7 days after operation,all sensitized dogs lost their graft functions,and the pathologic examination showed that their renal allografts were seriously rejected.2 of 3 dogs in control group also had acute rejection.Conclusion Repeated stimulation of canine lymphocyte may induce specific immune sensitization in canine recipients.Sensitized dogs have earlier rejection against renal allografts and a more deteriorated graft function.