Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy.
Manufacturing validation of biologically functional T cells targeted to CD19 antigen for autologous adoptive cell therapy.
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DOI：
10.1097/cji.0b013e318194a6e8
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发表时间：
2009-02
期刊：
影响因子：
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通讯作者：
Rivière I
中科院分区：
文献类型：
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作者：
Hollyman D;Stefanski J;Przybylowski M;Bartido S;Borquez-Ojeda O;Taylor C;Yeh R;Capacio V;Olszewska M;Hosey J;Sadelain M;Brentjens RJ;Rivière I
基于有前景的临床前数据，这些数据表明在重症联合免疫缺陷 beige小鼠模型中体内通过靶向CD19的T淋巴细胞根除全身性B细胞恶性肿瘤，我们正在慢性淋巴细胞白血病（CLL）和急性淋巴细胞白血病（ALL）患者中开展1期临床试验。在此我们展示了我们为生产和扩增源自CLL患者的临床级自体T细胞所开发的生物工艺的验证。我们证明了用源自莫洛尼鼠白血病病毒的复制缺陷型γ逆转录病毒载体进行基因修饰的T细胞，该载体编码一种靶向CD19（1928z）的嵌合抗原受体（CAR），可以用Dynabeads® CD3/CD28进行扩增。这种生物工艺使我们能够在使用Wave生物反应器的大规模半封闭培养系统中在大约2到3周内产生临床剂量的1928z + T细胞。这些1928z + T细胞不仅在体外，而且在携带播散性肿瘤的重症联合免疫缺陷beige小鼠体内仍然具有生物学功能。对于使用来自CLL患者的单采血液成分产品进行的所有四次验证运行，在T细胞扩增、用1928z CAR对T细胞进行转导、生物学活性、质量控制检测和放行标准方面的验证要求均已满足。此外，在T细胞扩增之后，偏态的Vβ T细胞受体库的多样性得到了显著恢复。这个经过验证的工艺将用于化疗难治性CLL患者和复发ALL患者的1期临床试验。它也可适用于涉及使用任何嵌合抗原受体或T细胞受体对患者或供体T细胞进行扩增和转导的其他临床试验。
Based on promising pre-clinical data demonstrating the eradication of systemic B cell malignancies by CD19-targeted T lymphocytes in vivo in SCID beige mouse models, we are launching Phase 1 clinical trials in patients with chronic lymphocytic leukemia (CLL) and acute lymphoblastic leukemia (ALL). We present here the validation of the bioprocess we developed for the production and expansion of clinical grade autologous T cells derived from patients with CLL. We demonstrate that T cells genetically modified with a replication-defective gammaretroviral vector derived from the Moloney murine leukemia virus encoding a chimeric antigen receptor (CAR) targeted to CD19 (1928z) can be expanded with Dynabeads® CD3/CD28. This bioprocess allows us to generate clinical doses of 1928z+ T cells in approximately 2 to 3 weeks in a large-scale semi-closed culture system using the Wave bioreactor. These 1928z+ T cells remain biologically functional not only in vitro but also in SCID beige mice bearing disseminated tumors. The validation requirements in terms of T cell expansion, T cell transduction with the 1928z CAR, biological activity, quality control testing and release criteria were met for all four validation runs using apheresis products from patients with CLL. Additionally, following expansion of the T cells, the diversity of the skewed Vβ T cell receptor repertoire was significantly restored. This validated process will be used in phase I clinical trials in patients with chemo-refractory CLL and in patients with relapsed ALL. It can also be adapted for other clinical trials involving the expansion and transduction of patient or donor T cells using any chimeric antigen receptor or T cell receptor.