Children's Hospital, University Hospital Tübingen, Tübingen, Germany.
Cytotherapy. 2013 Oct;15(10):1253-8. doi: 10.1016/j.jcyt.2013.05.014.
The CliniMACS device (Miltenyi Biotec, Bergisch Gladbach, Germany) was used for depletion of T-cell receptor alpha/beta positive (TCRαβ(+)) and CD19 positive (CD19(+)) cells from apheresis products.
Investigators performed 102 separations. Apheresis products with a median 5.8 (minimum to maximum, 1.2-10.4) × 10(10) mononuclear cells were used with a median 358 (92-1432) × 10(6) CD34(+) cells. There were 24.8% (6.1-45.7%) median TCRαβ(+) cells and 4.4% (1.2-11.7%) median B cells in the apheresis product.
After depletion, a median 0.00097% (0.00025-0.0048%) of TCRαβ(+) cells could be detected, and B cells, as determined as CD20(+) cells, were reduced to 0.0072% (0.0008-0.072%). TCRαβ(+) cells were depleted by log 4.7 (3.8-5.5), and B cells were depleted by log 4.1 (3.0-4.7). Recovery of mononuclear cells was 55% (33-77%), and recovery of CD34(+) cells was 73% (43-98%). Recovery of CD56(+)/3(-) natural killer cells was 80% (35-142%), recovery of TCR gamma/delta positive (TCRγδ(+)) T cells was 83% (39-173%) and recovery of CD14(+) cells was 79% (22-141%). Viability of cells was 98% (93-99%) after separation (all values median).
Profound depletion of TCRαβ(+) T cells can be achieved with the CliniMACS system. Recovery of CD34(+) stem cells is in the same range than after CD34(+) enrichment and CD3/CD19 depletion. Transplantation with >4 × 10(6) CD34(+) cells/kg can be performed for every patient with 1-5 × 10(4) TCRαβ(+) cells/kg and about 5-10 × 10(6) TCRγδ(+) cells/kg with two rounds of apheresis.
CliniMACS 设备（Miltenyi Biotec，Bergisch Gladbach，德国）用于从单采产品中去除 T 细胞受体 alpha/beta 阳性（TCRαβ(+））和 CD19 阳性（CD19(+））细胞。
研究人员进行了 102 次分离。使用中位数为 5.8（最小值至最大值，1.2-10.4）×10(10）单核细胞的单采产品，中位数为 358（92-1432）×10(6）CD34(+)细胞。单采产品中有 24.8%（6.1-45.7%）的中位数 TCRαβ(+）细胞和 4.4%（1.2-11.7%）的中位数 B 细胞。
在耗竭后，可检测到中位数为 0.00097%（0.00025-0.0048%）的 TCRαβ(+）细胞，作为 CD20(+)细胞确定的 B 细胞减少至 0.0072%（0.0008-0.072%）。TCRαβ(+）细胞的耗竭量为对数 4.7（3.8-5.5），B 细胞的耗竭量为对数 4.1（3.0-4.7）。单核细胞的回收率为 55%（33-77%），CD34(+)细胞的回收率为 73%（43-98%）。CD56(+）/3(-）自然杀伤细胞的回收率为 80%（35-142%），TCR 伽马/德尔塔阳性（TCRγδ(+））T 细胞的回收率为 83%（39-173%），CD14(+)细胞的回收率为 79%（22-141%）。分离后细胞的活力为 98%（93-99%）（所有值中位数）。
CliniMACS 系统可实现 TCRαβ(+）T 细胞的深度耗竭。CD34(+)干细胞的回收率与 CD34(+)富集和 CD3/CD19 耗竭后相同。对于每个患者，可以进行 >4×10(6）个 CD34(+)细胞/kg 的移植，每个患者可以使用 1-5×10(4）个 TCRαβ(+）细胞/kg 和 5-10×10(6）个 TCRγδ(+）细胞/kg，进行两轮单采。
