Myeloid cells in peripheral blood mononuclear cell concentrates inhibit the expansion of chimeric antigen receptor T cells.
Myeloid cells in peripheral blood mononuclear cell concentrates inhibit the expansion of chimeric antigen receptor T cells.
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DOI：
10.1016/j.jcyt.2016.04.003
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发表时间：
2016-07
期刊：
影响因子：
4.5
通讯作者：
Mackall CL
中科院分区：
文献类型：
--
作者：
Stroncek DF;Ren J;Lee DW;Tran M;Frodigh SE;Sabatino M;Khuu H;Merchant MS;Mackall CL
自体嵌合抗原受体（CAR）T细胞疗法已显示出有希望的临床结果，但T细胞产量一直是可变的。审查了CD 19-和GD 2-CAR T细胞生产记录，以确定变异性来源。CD 19-CAR T细胞用于治疗43例急性淋巴细胞白血病或淋巴瘤患者，GD 2-CAR T细胞用于治疗8例骨肉瘤患者和3例神经母细胞瘤患者。两种类型的CAR T细胞均使用自体外周血单核细胞（PBMC）浓缩物和抗CD 3/CD 28珠粒进行T细胞富集和模拟。制造的前6种GD 2-和前22种CD 19-CAR T细胞产物的比较显示，GD 2-CAR T细胞产物比CD 19-CAR T细胞产物含有更少的转导细胞（147 ± 102 × 106 vs 1502 ± 1066 × 106; P = 0.0059），其PBMC浓缩液含有更多的单核细胞（31.4 ± 12.4% vs 18.5 ± 13.7%; P = 0.019）。在生产的前28种CD 19-CAR T细胞产品中，有4种扩增效果不佳，每千克产生的转导T细胞少于1 × 106个。当将这4名患者的PBMC浓缩物与其他24名患者的PBMC浓缩物进行比较时，扩增不良产物的PBMC浓缩物含有更大量的单核细胞（39.8 ± 12.9%对15.3 ± 10.8%，P = 0.0014）。在CD 19-CAR T细胞扩增不良的患者中，使用冷冻保存的PBMC浓缩物重复制造两名患者，但合并单核细胞消耗塑料粘附步骤，并为两名患者生产足够剂量的CAR T细胞。CAR T细胞扩增的可变性至少部分是由于起始PBMC浓缩物被单核细胞污染。
Autologous chimeric antigen receptor (CAR) T-cell therapies have shown promising clinical outcomes, but T-cell yields have been variable. CD19- and GD2-CAR T-cell manufacturing records were reviewed to identify sources of variability. CD19-CAR T cells were used to treat 43 patients with acute lymphocytic leukemia or lymphoma and GD2-CAR T cells to treat eight patients with osteosarcoma and three with neuroblastoma. Both types of CAR T cells were manufactured using autologous peripheral blood mononuclear cells (PBMC) concentrates and anti-CD3/CD28 beads for T-cell enrichment and simulation. A comparison of the first 6 GD2- and the first 22 CD19-CAR T-cell products manufactured revealed that GD2-CAR T-cell products contained fewer transduced cells than CD19-CAR T-cell products (147 ± 102 × 106 vs 1502 ± 1066 × 106; P = 0.0059), and their PBMC concentrates contained more monocytes (31.4 ± 12.4% vs 18.5 ± 13.7%; P = 0.019). Among the first 28 CD19-CAR T-cell products manufactured, four had poor expansion yielding less than 1 × 106 transduced T cells per kilogram. When PBMC concentrates from these four patients were compared with the 24 others, PBMC concentrates of poorly expanding products contained greater quantities of monocytes (39.8 ± 12.9% vs. 15.3 ± 10.8%, P = 0.0014). Among the patients whose CD19-CAR T cells expanded poorly, manufacturing for two patients was repeated using cryopreserved PBMC concentrates but incorporating a monocyte depleting plastic adherence step, and an adequate dose of CAR T cells was produced for both patients. Variability in CAR T-cell expansion is due, at least in part, to the contamination of the starting PBMC concentrates with monocytes.