异基因造血干细胞移植治疗BXSB狼疮鼠

Treatment of BXSB With Allogeneic Hematopoietic Stem Cell Transplantation

目的探索异基因外周血造血干细胞移植治疗重型系统性红斑狼疮(SLE)的有效性、安全性。方法用小鼠粒细胞集落刺激因子动员、采集供者C57BL/6的正常造血干细胞,输入经用环磷酰胺(CTX)、马利兰预处理的BXSB狼疮鼠体内,用血细胞分析仪和流式细胞仪动态观察移植后BXSB狼疮鼠的造血和免疫重建,尿蛋白试剂、ELISA法、免疫荧光法检测移植后的BXSB鼠尿蛋白、抗双链DNA抗体、肾病理改变和观察与移植相关的并发症、死亡率,并用微卫星PCR法检测移植后BXSB狼疮鼠的植活情况。结果移植后60dDNA指纹图分析BXSB狼疮鼠转为供者的DNA指纹图,移植后白细胞下降至最低值是(0.2±0.1)×109/L,白细胞从最低值上升至1.0×109/L的时间是移植后(47.5±12.8)d,移植后第8周CD3-CD5+和CD3+CD5+尚未恢复,移植后40d95.5%BXSB狼疮鼠尿蛋白减少,90.9%肾荧光减少,与传统的治疗方法比较差异有显著性,54.5%抗双链DNA抗体转阴,明显高于传统治疗组,但差异无显著性。与移植相关的出血、体重减轻、肺部感染及死亡率与传统的治疗方法比较差异均无显著性。结论Allo-HSCT是一种有效的较彻底的治疗重型SLE的方法,其在降低BXSB狼疮鼠尿蛋白,使抗双链DNA抗体转阴及改善肾病理改变方面明显优于传统的治疗方法,但与移植相关的并发症、死亡率较高。

Objective To investigate the efficacy,safety of allogeneic hematopoietic stem cell transplantation(Allo-HSCT)in lupus mice.Methods22BXSB was pre-treated with myleran and cyclophosphamide,12BXSB were treated with CTX and prednisone as the control group.Peripheral blood stem cells of doner mouse(C57BL/6)after mobilizing with G-CSF were infused into recipients after pretreatment.Hematopoietic and immune reconstitution after transplantation were measured by blood cell analysis and flowcytometry.Recipients' renal pathological changes were measured with direct immunofluorescence after transplantation,proteinuria and anti-dsDNA antibody were also determined.Complications and mortality were compared between the two groups.Recipients'genetic map was analysed with PCR.Results Recipients'complete chimerism was observed after60days of transplantation.The lowest value of WBC declined to0.2±0.1×10 9 /L in Allo-HSCT group.The duration of WBC increased from the lowest value up to1.0×10 9 /L was47.5±12.8d in Allo-HSCT group.Early immune reconstitution could not be found in Allo-HSCT group.After transplantation reduction of proteinuria occurred in95.5%BXSB,anti-ds-DNA antibody turned to negative in54.5%BXSB,decrease of kindey immunofluorescence in90.9%BXSB,there was a significant difference between the two groups.There was no significant difference in the incidences of complication of bleeding,pneumonia and mortality between the Allo-HSCT group and the control group.Conclusions Allo-HSCT is an effective treatment method for lupus mice.It may be helpful in improving proteinuria and anti-dsDNA antibody and renal pathological changes of lupus mice.But the incidences of complication and mortality related to transplantation in Allo-HSCT group are higher than those in control group,so the value of Allo-HSCT in the treatment lupu mice has to be further studied.