Link H, Arseniev L, Bähre O, Berenson R J, Battmer K, Kadar J G, Jacobs R, Casper J, Kühl J, Schubert J, Diedrich H, Poliwoda H
Department of Hematology and Oncology, Medizinische Hochschule Hannover, Germany.
Blood. 1995 Oct 1;86(7):2500-8.
Allogeneic peripheral blood progenitor cells (PBPCs) were transplanted after immunoselection of CD34+ cells. Two patient groups were studied: group I patients received immunoselected blood CD34+ cells and unmanipulated marrow cells from the same donor. Group II patients were given immunoselected blood and bone marrow (BM) CD34+ cells. One to 6 weeks before bone marrow transplantation (BMT), PBPCs from HLA-identical and MLC- sibling donors were mobilized with granulocyte colony-stimulating factor (G-CSF) (5 micrograms/kg twice daily subcutaneously) for 5 days. Aphereses were performed at days 4 and 5 of G-CSF application. CD34+ cells were separated from the pooled PBPC concentrates by immunoadsorption onto avidin with the biotinylated anti-CD34 monoclonal antibody 12.8 and then stored in liquid nitrogen. BM was procured on the day of transplantation. Patients were conditioned with either busulfan (16 mg/kg) or total body irradiation (12 Gy) followed by cyclophosphamide (120 mg/kg). Cyclosporin A and short methotrexate were used for graft-versus-host disease (GVHD) prophylaxis. After transplantation, all patients received 5 micrograms G-CSF/kg/d from day 1 until greater than 500 neutrophils/microL were reached and 150 U erythropoietin/kg/d from day 7 until erythrocyte transfusion independence for 7 days. Group I consisted of patients with acute myeloid leukemia (AML) (n = 2), chronic myeloid leukemia (CML) (n = 2), and T-gamma-lymphoproliferative syndrome and BM aplasia (n = 1). The patients received a mean of 3.3 x 10(6) CD34+ and 3.7 x 10(5) CD3+ cells/kg body weight of PBPC origin and 4.5 x 10(6) CD34+ and 172 x 10(5) cells/kg body weight of BM origin. Group II consisted of five patients (two AML, two CML, one non-Hodgkin's lymphoma). They received a mean of 3.3 x 10(6) CD34+ and 3.2 x 10(5) CD3+ cells/kg from PBPC and 1.4 x 10(6) CD34+ and 0.6 x 10(5) CD3+ cells from BM. A matched historical control group (n = 12) transplanted with a mean of 5.2 x 10(6) CD34+ and 156 x 10(5) CD3+ cells/kg from BM alone was assembled for comparison. In group I, the median time to neutrophil recovery to > 100, > 500, and > 1,000/microL was 12, 15, and 17 days, respectively. Patients from group II reached these neutrophil levels at days 13, 15 and 17 post BMT. Neutrophil recovery in the control patient group occurred at days 17, 18, and 20 respectively. Group I patients were given platelet transfusions within 18 days and red blood cells within 10 days, whereas for group II patients, these time points were 26 and 17 days, respectively. These same transfusions could be ceased within 38 and 24 days, respectively, in control patients. The addition of about 2% more peripheral blood CD3+ cells (group I patients) did not result in higher grades of acute GVHD (median grade II) as compared with the controls (median grade II). Four of five group II patients showed no signs of acute GVHD. These data suggest that the addition of immunoselected allogeneic CD34+ progenitor cells to BM cells may accelerate hematopoietic recovery.
在对CD34+细胞进行免疫选择后，移植了异基因外周血祖细胞（PBPCs）。研究了两个患者组：第一组患者接受了来自同一供体的经免疫选择的血液CD34+细胞和未处理的骨髓细胞。第二组患者接受了经免疫选择的血液和骨髓（BM）CD34+细胞。在骨髓移植（BMT）前1至6周，用粒细胞集落刺激因子（G-CSF）（5微克/千克，每日皮下注射两次）动员来自HLA相同和混合淋巴细胞培养阴性同胞供体的PBPCs，共5天。在应用G-CSF的第4天和第5天进行血细胞分离。通过用生物素化抗CD34单克隆抗体12.8免疫吸附到抗生物素蛋白上，从汇集的PBPC浓缩物中分离出CD34+细胞，然后储存在液氮中。在移植当天采集骨髓。患者接受白消安（16毫克/千克）或全身照射（12 Gy）预处理，随后给予环磷酰胺（120毫克/千克）。使用环孢素A和短疗程甲氨蝶呤预防移植物抗宿主病（GVHD）。移植后，所有患者从第1天开始接受5微克G-CSF/千克/天，直至中性粒细胞计数大于500/微升，从第7天开始接受150单位促红细胞生成素/千克/天，直至红细胞输注独立7天。第一组包括急性髓性白血病（AML）患者（n = 2）、慢性髓性白血病（CML）患者（n = 2）以及T-γ淋巴细胞增殖综合征和骨髓发育不全患者（n = 1）。这些患者接受的PBPC来源的CD34+细胞平均为3.3×10⁶/千克体重，CD3+细胞平均为3.7×10⁵/千克体重，BM来源的CD34+细胞平均为4.5×10⁶/千克体重，细胞平均为172×10⁵/千克体重。第二组包括五名患者（两名AML、两名CML、一名非霍奇金淋巴瘤）。他们接受的PBPC来源的CD34+细胞平均为3.3×10⁶/千克，CD3+细胞平均为3.2×10⁵/千克，BM来源的CD34+细胞平均为1.4×10⁶，CD3+细胞平均为0.6×10⁵。组建了一个匹配的历史对照组（n = 12），该组仅移植了平均5.2×10⁶/千克的BM来源的CD34+细胞和156×10⁵/千克的CD3+细胞用于比较。在第一组中，中性粒细胞恢复至>100/微升、>500/微升和>1000/微升的中位时间分别为12天、15天和17天。第二组患者在BMT后第13天、15天和17天达到这些中性粒细胞水平。对照患者组中性粒细胞恢复分别发生在第17天、18天和20天。第一组患者在18天内接受血小板输注，在10天内接受红细胞输注，而第二组患者，这些时间点分别为26天和17天。对照患者分别在38天和24天内可以停止这些相同的输血。与对照组（中位II级）相比，第一组患者外周血CD3+细胞增加约2%并没有导致更高等级的急性GVHD（中位II级）。五名第二组患者中有四名没有急性GVHD的迹象。这些数据表明，向BM细胞中添加经免疫选择的异基因CD34+祖细胞可能会加速造血恢复。
