Urbano-Ispizua A, Solano C, Brunet S, de la Rubia J, Odriozola J, Zuazu J, Figuera A, Caballero D, Martínez C, García J, Sanz G, Torrabadella M, Alegre A, Pérez-Oteiza J, Jurado M, Oyonarte S, Sierra J, García-Conde J, Rozman C
Bone Marrow Transplant. 1998 Sep;22(6):519-25. doi: 10.1038/sj.bmt.1701386.
The objective of this study was to analyze CD34+ cell recovery and T cell depletion (TCD) achieved in CD34+ cell grafts using either immunoadsorption or immunomagnetic methods applied to leukapheresis products from healthy donors. We also wanted to determine the kinetics of engraftment and incidence and severity of graft-versus-host disease (GVHD) after allogeneic transplantation of selected CD34+ cells. HLA-identical sibling donors received G-CSF. After leukapheresis, peripheral blood progenitor cells were selected using immunoadsorption (Ceprate SC) (n = 38) or immunomagnetic (Isolex 300) (n = 24) methods. Sixty-two patients, with a median age of 42 years (range 17-60) diagnosed with hematological malignancies were conditioned with either cyclophosphamide and total body irradiation (n = 43) or busulphan and cyclophosphamide (n = 19). GVHD prophylaxis consisted of cyclosporin A (CsA) and prednisone (n = 48), CsA alone (n = 11) and CsA and methotrexate (n = 3). The median yield and purity of CD34+ cells after the procedure was 65 and 66% with immunoadsorption, and 48 and 86% with immunomagnetic method, respectively. The median number (range) of CD34+ cells infused into the patients was 3.5 x 10(6)/kg (1-9.6). The median number (range) of CD3+ cells administered was 0.4 x 10(6)/kg (0.01-2) using immunoadsorption and 0.14 x 10(6)/kg (0.03-2.5) using immunomagnetic methods. Neutrophil recovery >500 and >1000/microl was achieved at a median (range) of 13 days (8-22) and 14 days (9-31), respectively. Platelets recovered to >20000 and >50000/microl at a median (range) of 13 days (0-128) and 18 days (0-180), respectively. Two patients developed graft failure. Acute GVHD in patients at risk was clinical grade 0 (n = 43), I (n = 8), II (n = 4) and III (n = 1). No patient developed acute GVHD grade IV. Chronic GVHD was limited in two cases and extensive in four cases. The actuarial probability of acute GVHD II-IV was 10% (95% CI, 1-19%), and of extensive chronic GVHD was 12% (95% CI, 11-13%). The cumulative incidence of transplant-related mortality was 12.6%, and this figure was 9% at 6 months. In conclusion, with the immunomagnetic procedure, a lower recovery and higher purity of CD34+ cells, and stronger TCD is obtained as compared to immunoadsorption (P = 0.008, P < 0.0001 and P = 0.0002, respectively). Our results also indicate that allogeneic transplantation of selected CD34+ cells is associated with a very rapid engraftment and with a low incidence of severe GVHD.
本研究的目的是分析使用免疫吸附或免疫磁珠方法对健康供者白细胞单采产物进行处理后,CD34+细胞移植物中的CD34+细胞回收率和T细胞清除(TCD)情况。我们还想确定选定的CD34+细胞进行异基因移植后植入的动力学以及移植物抗宿主病(GVHD)的发生率和严重程度。HLA相同的同胞供者接受粒细胞集落刺激因子(G-CSF)。白细胞单采后,使用免疫吸附法(Ceprate SC)(n = 38)或免疫磁珠法(Isolex 300)(n = 24)选择外周血祖细胞。62例诊断为血液系统恶性肿瘤的患者,中位年龄42岁(范围17 - 60岁),接受了环磷酰胺和全身照射(n = 43)或白消安和环磷酰胺(n = 19)进行预处理。GVHD预防方案包括环孢素A(CsA)和泼尼松(n = 48)、单独使用CsA(n = 11)以及CsA和甲氨蝶呤(n = 3)。该操作后CD34+细胞的中位回收率和纯度,免疫吸附法分别为65%和66%,免疫磁珠法分别为48%和86%。输注给患者的CD34+细胞中位数量(范围)为3.5×10⁶/kg(1 - 9.6)。使用免疫吸附法给予的CD3+细胞中位数量(范围)为0.4×10⁶/kg(0.01 - 2),使用免疫磁珠法为0.14×10⁶/kg(0.03 - 2.5)。中性粒细胞恢复>500/μl和>1000/μl的中位时间(范围)分别为13天(8 - 22天)和14天(9 - 31天)。血小板恢复到>20000/μl和>50000/μl的中位时间(范围)分别为13天(0 - 128天)和18天(0 - 180天)。2例患者发生移植失败。有风险的患者中急性GVHD的临床分级为0级(n = 43)、I级(n = 8)、II级(n = 4)和III级(n = 1)。无患者发生IV级急性GVHD。慢性GVHD 2例为局限性,4例为广泛性。急性GVHD II - IV级的精算概率为10%(95%CI,1 - 19%),广泛性慢性GVHD的精算概率为12%(95%CI,11 - 13%)。移植相关死亡率的累积发生率为12.6%,6个月时为9%。总之,与免疫吸附法相比,免疫磁珠法获得的CD34+细胞回收率较低但纯度较高,TCD更强(分别为P = 0.008、P < 0.0001和P = 0.0002)。我们的结果还表明,选定的CD34+细胞进行异基因移植后植入非常迅速,严重GVHD的发生率较低。