Tichelli A, Gratwohl A, Bargetzi M, Nissen C, Wernli M, Herrmann R, Orth B, Signer E, Speck B
Departement Zentrallaboratorium, Universittsspital, Basel.
Schweiz Med Wochenschr. 1996 Feb 10;126(6):201-6.
Peripheral blood is increasingly used instead of bone marrow as a source of hemopoietic precursor cells for transplantation. The optimal technique still needs to be defined. Selection of CD34+ cells in transplant material may be of benefit in allogeneic and autologous peripheral blood precursor cell transplantation (PBPCT), since it allows elimination of unwanted CD34-negative cells, such as T-cells and contaminating tumor cells. We have evaluated the feasibility of CD34 selection in PB transplants and studied hemopoietic reconstitution after autologous transplantation of CD34 selected precursor cells. Between August 1994 and June 1995 CD34 selection was performed on 12 transplants for 9 patients with malignant disease (non-Hodgkin lymphoma [n = 5]; Ewing sarcoma [n = 1]; chronic lymphocytic leukemia [n = 1]; breast cancer [n = 1]; multiple myeloma [n = 1]). PBPC were collected with a Fenwall CS 3000 harvester after stimulation with G-CSF. For selection of CD34+ cells the Ceprate LC34 system (CellPro) was used. A median CD34 purity of 73% (range 40-94%) was achieved. The median number of CD34 positive cells per transplant was 4.8 x 10(6)/kg body weight (range 0.7-15.8). The median number of colony forming cells per transplant was 31 x 10(4)/kg body weight (range 1.5-131.3). For autologous PBPCT the minimal number of CD34 positive cells required in the transplantate was arbitrarily set at 1.0 x 10(6)/kg body weight. This number was achieved in 10 of the 12 transplants. The median loss of CD34+ cells during selection was 1.5 x 10(6)/kg body weight (range 0.2-6.4). In 2 patients the total number was reduced to below the critical value of 1.0 x 10(6)/kg. 7 of the 9 patients received the CD34 selected transplant after intensive chemotherapy and irradiation. The median follow-up time after PBPCT was 196 days (range 62-278). All 7 patients are now alive and with normal hemopoietic function. A granulocyte count above 0.5 x 10(9)/l and a platelet count above 20 x 10(9)/l was achieved on day 14 (median), and on day 19 after PBPCT. We conclude that CD34 selection is technically feasible and that CD34 selected cells can be used for PBPCT. The procedure is time consuming and expensive; it requires complex organization at laboratory level, and the benefit of CD34 selection with regard to T-cell depletion and tumor purging still needs to be proven. However, CD34+ selection is likely to open new perspectives in transplantation medicine.
外周血越来越多地被用作移植造血前体细胞的来源，而不再使用骨髓。最佳技术仍有待确定。在同种异体和自体外周血前体细胞移植（PBPCT）中，选择移植材料中的CD34+细胞可能有益，因为这样可以去除不需要的CD34阴性细胞，如T细胞和污染的肿瘤细胞。我们评估了在PB移植中选择CD34的可行性，并研究了自体移植经CD34选择的前体细胞后的造血重建情况。1994年8月至1995年6月期间，对9例恶性疾病患者的12次移植进行了CD34选择（非霍奇金淋巴瘤[n = 5]；尤因肉瘤[n = 1]；慢性淋巴细胞白血病[n = 1]；乳腺癌[n = 1]；多发性骨髓瘤[n = 1]）。用G-CSF刺激后，使用Fenwall CS 3000血细胞分离机采集PBPC。为了选择CD34+细胞，使用了Ceprate LC34系统（CellPro）。CD34的中位纯度达到73%（范围40 - 94%）。每次移植的CD34阳性细胞中位数为4.8×10⁶/kg体重（范围0.7 - 15.8）。每次移植的集落形成细胞中位数为31×10⁴/kg体重（范围1.5 - 131.3）。对于自体PBPCT，移植产物中所需的CD34阳性细胞的最小数量被任意设定为1.0×10⁶/kg体重。12次移植中有10次达到了这个数量。选择过程中CD34+细胞的中位损失为1.5×10⁶/kg体重（范围0.2 - 6.4）。2例患者的总数降至低于1.0×10⁶/kg的临界值。9例患者中有7例在强化化疗和放疗后接受了经CD34选择的移植。PBPCT后的中位随访时间为196天（范围62 - 278）。所有7例患者目前均存活且造血功能正常。在PBPCT后的第14天（中位数）和第19天，粒细胞计数高于0.5×10⁹/L，血小板计数高于20×10⁹/L。我们得出结论，选择CD34在技术上是可行的，并且经CD34选择的细胞可用于PBPCT。该过程耗时且昂贵；它需要实验室层面的复杂组织安排，并且CD34选择在T细胞清除和肿瘤净化方面的益处仍有待证实。然而，选择CD34+可能会为移植医学开辟新的前景。
