应用树突状细胞单克隆抗体预防脾移植排斥反应的实验研究

Research on the prevention of splenic graft rejection in vitro with anti-dendritic cell monoclonal antibody

目的 建立治疗性单克隆抗体溶液持续器官灌注模型 ,应用抗大鼠树突状细胞单克隆抗体 (WZD)降低大鼠移植脾的免疫原性。方法 以WZD溶液持续灌注移植脾 ,筛选最佳方案 ,并观察灌注后混合淋巴细胞培养(mixedlymphocyteculture ,MLC)中淋巴细胞的增殖情况及灌注后移植脾细胞的超微结构。结果 WZD灌注组MLC淋巴细胞增殖能力明显降低 ,与对照组及应用环孢霉素A(CyclosporinA ,CsA)组相比差异显著 (P <0 .0 5 ) ,与WZD灌注 +CsA组相比无明显差异 (P >0 .0 5 ) ,2 1℃灌注组移植脾细胞功能活跃 ,线粒体损伤小。结论 移植脾在 2 1℃下 ,每小时给予灌注 5 0ml单抗溶液获得了最佳效果 ,WZD溶液灌注移植脾 ,可去除树突状细胞 ,确能降低其免疫原性 ,有可能减少免疫抑制剂的应用剂量。

Objective To reduce immunogenicity of the donor spleen,a rat model of continuous perfusion of graft was established with therapeutic anti dendritic cells monoclonal antibody(WZD).Methods We perfused donor rat spleens continuously with WZD and selected the best protocol among them,then we examined lymphopoiesis by mixed lymphocyte culture (MLC) and observed ultrastructure of donor splenocytes.Results Perfusion alone significantly depressed the ability of lymphopoiesis( P <0.05,compared with control group and Cyclosporine A treatment alone).WZD perfusion together with Cyclosporine A treatment exhibited the same impact( P >0.05).The function of splenocyte in graft which perfused at 21℃ was more active,and the mitochondrial injury was slight.Conclusion The best effect can be obtained by perfusion with WZD at a speed of 50ml/h at 21℃.Pretransplant perfusion of the spleen with WZD may reduce the immunogenicity of the graft and the dosage of immunosuppressant.