Locatelli F, Perotti C, Torretta L, Maccario R, Montagna D, Ravelli A, Giorgiani G, De Benedetti F, Giraldi E, Magnani M L, De Stefano P, Martini A
Dipartimento di Scienze Pediatriche, Università di Pavia, IRCCS Policlinico San Matteo, P.le Golgi 2, 27100 Pavia, Italy.
Haematologica. 1999 Sep;84(9):839-43.
Autologous transplant of lymphocyte-depleted peripheral blood stem cells has been proposed for treatment of patients with severe autoimmune disease. However, until now, no data are available on the safety and feasibility of both stem cell collection and selection in pediatric patients with these disorders. We report on three children affected by systemic sclerosis with lung involvement, who received chemotherapy and granulocyte colony-stimulating factor (G-CSF) to mobilize autologous peripheral blood progenitors.
The priming regimen consisted of cyclophosphamide (CY, 4 g/m(2)) and G-CSF (lenograstim, 10 microg/kg/day starting 2 days after cyclophosphamide administration until stem cell collection). Leukapheresis was performed when WBC and CD34+ cell count were at least 2 x 10(9)/L and 0.03 x 10(9)/L, respectively. In the first patient, positive selection of CD34+ cells was performed through the Ceprate SC stem cell concentrator (CellPro, Bothell, WA, USA). In the remaining 2 children, progenitor cells were also purged with negative selection of CD4+ and CD8+ lymphocytes performed by means of the Isolex 300i device (Baxter).
All patients tolerated the priming regimen well and did not present any sign of autoimmune disease exacerbation. Collection was successful in all children and the number of CD34+ cells before selection ranged between 10.7 x 10(6) and 17.6 x 10(6)/kg of patient body weight. The selection of haematopoietic stem cells in the 3 patients resulted in at least 2. 6-log T-cell depletion of the cell content, with a recovery of the initial value of CD34+ cells comprised between 21 and 44%. After, a preparative regimen consisting of CY (200 mg/kg over 4 days) and Campath-1 G in vivo (10 mg/day for 2 consecutive days), patients were transplanted using cryopreserved lymphocyte-depleted progenitor cells. In all cases, a prompt hematopoietic engraftment was observed.
Taken together these data suggest that mobilization, collection and selection of hematopoietic progenitors are safe and feasible in children with autoimmune disease.
已有人提出采用去除淋巴细胞的自体外周血干细胞移植来治疗重症自身免疫性疾病患者。然而，迄今为止，尚无关于患有这些疾病的儿科患者干细胞采集和分选的安全性及可行性的数据。我们报告了3例患有系统性硬化症并伴有肺部受累的儿童，他们接受了化疗及粒细胞集落刺激因子（G-CSF）以动员自体外周血祖细胞。
预处理方案包括环磷酰胺（CY，4 g/m²）和G-CSF（来格司亭，在环磷酰胺给药后2天开始，10 μg/kg/天，直至干细胞采集）。当白细胞计数和CD34⁺细胞计数分别至少达到2×10⁹/L和0.03×10⁹/L时，进行白细胞分离术。在第1例患者中，通过Ceprate SC干细胞富集仪（CellPro，美国华盛顿州博塞尔）对CD34⁺细胞进行阳性分选。在其余2名儿童中，还通过Isolex 300i装置（百特）对CD4⁺和CD8⁺淋巴细胞进行阴性分选来清除祖细胞。
所有患者对预处理方案耐受性良好，且未出现自身免疫性疾病加重的任何迹象。所有儿童的采集均成功，分选前CD34⁺细胞数量在10.7×10⁶至17.6×10⁶/kg患者体重之间。3例患者的造血干细胞分选导致细胞成分中T细胞至少减少2.6个对数级，CD34⁺细胞初始值的恢复率在21%至44%之间。之后，采用由CY（4天内200 mg/kg）和体内Campath-1 G（连续2天，10 mg/天）组成的预处理方案，使用冷冻保存的去除淋巴细胞的祖细胞对患者进行移植。在所有病例中，均观察到迅速的造血植入。
综合这些数据表明，造血祖细胞的动员、采集和分选在患有自身免疫性疾病的儿童中是安全可行的。
