儿童难治性自身免疫性疾病自体外周血干细胞动员采集和分选的临床研究

目的 探讨儿童难治性自身免疫性疾病进行自体外周血干细胞动员采集的安全性和CD34+细胞分选纯化的可行性及其临床意义.方法 8例儿童难治性自身免疫性疾病,包括4例系统性红斑狼疮、2例皮肌炎,1例幼年型类风湿关节炎和1例多发性硬化,予行CD34+细胞纯化的自体外周血干细胞移植.首先采用环磷酰胺(CTX)联合粒细胞集落刺激因子(G-CSF)方案动员外周血干细胞,然后采用CS-3000血细胞分离机采集外周血,通过CliniMACS细胞分选仪分选自体外周血CD34+细胞,将其用保养液配置冻存于-80℃冰箱.采用非清髓内去除T的预处理方案,即卡氮芥+足叶乙苷+阿糖胞苷+马法兰+抗胸腺球蛋白(ATG)或CTX+ATG或CTX+马法兰+ATG,于第0天回输自体外周血CD34+细胞.结果 儿童能够耐受自体外周血干细胞动员采集过程,无动员相关死亡,动员后获得的单个核细胞数和CD34+细胞数的平均值分别为8.35×108/kg和7.92×106/kg,纯化后的白体外周血CD34+和CD3+细胞数的平均值分别为6.28×106/kg和0.71×105/kg.回输后中性粒细胞和血小板的植入中位时间分别为+11d和+15 d.结论 经CTX联合G-CSF方案可动员出足量的外周血干细胞,经CS-3000血细胞分离机采集可获得足够的单个核细胞,在动员过程中原发病无明显进展恶化,患儿能耐受动员方案,采集过程顺利安全;经CliniMACS细胞分选仪分选的自体外周血CD34+细胞纯度高,移植后造血恢复;采用CD34+细胞纯化的自体外周血移植治疗是常规治疗无效的儿童难治性自身免疫性疾病的可选择治疗措施之一.

A study of the mobilization, collection and selection of autologous peripheral blood stem cells in patients with autoimmune diseases undergoing autologous hematopoietic stem cell transplantation in juvenile severe autoimmune disease

Objective To explore the safety of mobilization and collection as well as the feasibility of selection of autologous peripheral blood stem cells (auto-PBSC) from patients with juvenile severe autoimmune diseases (AID) for autologous hematopoietic stem cell transplantation (auto-HSCT). The clinical significance of these procedure is evaluated. Methods Eight patients with AID, including four patients with systemic lupus erythematosus(SLE),two patients with dermatomysoitis, one patient with juvenile rheumatoid arthritis (JRA), one patient with multiple sclerosis(MS),underwent auto-HSCT. Auto-PBSCs were mobilized in 8 patients using cyclophosphamide(CTX) and granulocyte colony-stimulating factor (G-CSF), and their PBSCs were collected by CS-3000 Blood Cell Separator, then the CD34+cells were selected and purified by CliniMACS CD34+cell selection device. The CD34+ cells were frozenand preserved under -80 ℃ ALL patients received non-myeloablative or lymphoablative conditioning regimens which consisted of CTX/Mel/ATG or CTX/ATG or BEAM/ATG. All patient received CD34+ cells transplantation. The safety of mobilization and collection process of auto-PBSC as well asthe feasibility of selection and purification of CD34+cells were recorded and hematopoietic reconstruction were evaluated. Results All patients tolerated the collection process well, and there was no mobilization-related mortality. The number of collected MNCs and CD34+ cells were 8.35×108/kg and 7.92×106/kg respectively. The number of CD34+ and CD3+ cells after purification was 6.28×106/kg and0.71 ×105/kg respectively. The mean granulocytes and platelet engraftment occurred on days 11 and 15 after G-CSF regimen, and they can be collected using CS-3000 instrument. PBSC mobilization and collection from patients with juvenile severe AID is safe. The CD34+ cell can be highly purified. The auto-PBSC CD34+cell transplantation is an alternative therapy for severe AIDs that do not respond to conventional treatments.