脾移植前去除成熟树突状细胞预防移植排斥反应的体外研究

In vitro study on prevention of graft rejection with pretreatment of donor spleen by elimination of dendritic cells

目的 建立治疗性单抗溶液持续灌注大鼠脾脏模型 ,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)淋巴细胞增殖情况及脾细胞的超微结构 ,以及灌注后移植脾细胞的免疫组化情况。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CsA组相比差异有显著意义 (P <0 0 5 ) ,与WZD灌注 +CsA组相比差异无明显意义 (P >0 .0 5 ) ,2 1℃灌注组脾细胞功能活跃 ,线粒体损伤小 ,灌注后移植脾DC呈现强荧光染色。结论 移植脾在 2 1℃下 ,每小时给予灌注 5 0ml单抗溶液灌注获得了最佳效果 ,WZD溶液灌注移植脾 ,可去除成熟树突状细胞 ,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, we established a rat model of continuous perfusion of spleen with therapeutic anti dendritic cell monoclonal antibody (WZD). Methods The donor rat spleen was continuously perfused with WZD to select the best protocol. Then the lymphopoiesis was examined with mixed lymphocyte culture (MLC) and the splenic cells were observed microscopically and immunohistochemically. Results Lymphopoiesis was significantly inhibited in WZD perfused group as compared with the control and cyclosporine A (CsA) treated groups (P<0.05). WZD perfusion in combination with CsA treatment exerted the same effects as those of WZD perfusion alone (P>0.05). The function of splenocyte in the grafts perfused at 21℃ was more active and the mitochondrial injury was slight. Conclusions The best effect can be obtained by perfusion with WZD at a speed of 50 ml/h at 21℃. Perfusion of the spleen with WZD before transplantation may reduce the immunogenicity of the graft and the dosage of immunosuppressive agents.