Adoptive immunotherapy is a new potential method of tumour therapy, among which anti‐CD19 chimeric antigen receptor T‐cell therapy (CAR‐T cell), is a typical treatment agent for haematological malignancies. Previous clinical trials showed that the quality and phenotype of CAR‐T cells expanded ex vivo would seriously affect the tumour treatment efficacy. Although magnetic beads are currently widely used to expand CAR‐T cells, the optimal expansion steps and methods have not been completely established. In this study, the differences between CAR‐T cells expanded with anti‐CD3/CD28 mAb‐coated beads and those expanded with cell‐based aAPCs expressing CD19/CD64/CD86/CD137L/mIL‐15 counter‐receptors were compared. The results showed that the number of CD19‐specific CAR‐T cells with a 4‐1BB and CD28 co‐stimulatory domain was much greater with stimulation by aAPCs than that with beads. In addition, the expression of memory marker CD45RO was higher, whereas expression of exhausted molecules was lower in CAR‐T cells expanded with aAPCs comparing with the beads. Both CAR‐T cells showed significant targeted tumoricidal effects. The CAR‐T cells stimulated with aAPCs secreted apoptosis‐related cytokines. Moreover, they also possessed marked anti‐tumour effect on NAMALWA xenograft mouse model. The present findings provided evidence on the safety and advantage of two expansion methods for CAR‐T cells genetically modified by piggyBac transposon system.

中文翻译：

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用 aAPC 或磁珠离体扩增后 piggyBac 介导的抗 CD19 CAR-T 细胞的评估

过继性免疫疗法是一种潜在的肿瘤治疗新方法，其中抗CD19嵌合抗原受体T细胞疗法（CAR-T细胞）是血液系统恶性肿瘤的典型治疗药物。以往的临床试验表明，离体扩增的CAR-T细胞的质量和表型会严重影响肿瘤的治疗效果。虽然磁珠目前广泛用于扩增 CAR-T 细胞，但最佳扩增步骤和方法尚未完全确立。在这项研究中，比较了用抗 CD3/CD28 mAb 包被的珠子扩增的 CAR-T 细胞与用表达 CD19/CD64/CD86/CD137L/mIL-15 反受体的基于细胞的 aAPC 扩增的细胞之间的差异。结果表明，具有 4-1BB 和 CD28 共刺激结构域的 CD19 特异性 CAR-T 细胞的数量在 aAPC 的刺激下比在珠子的刺激下要多得多。此外，与珠子相比，用 aAPC 扩增的 CAR-T 细胞中记忆标记 CD45RO 的表达较高，而耗尽分子的表达较低。两种 CAR-T 细胞均显示出显着的靶向杀肿瘤作用。用 aAPC 刺激的 CAR-T 细胞分泌凋亡相关的细胞因子。此外，它们还对 NAMALWA 异种移植小鼠模型具有显着的抗肿瘤作用。目前的研究结果为两种扩增方法的安全性和优势提供了证据，这些方法用于转基因 CAR-T 细胞 而与珠子相比，用 aAPC 扩增的 CAR-T 细胞中耗尽分子的表达较低。两种 CAR-T 细胞均显示出显着的靶向杀肿瘤作用。用 aAPC 刺激的 CAR-T 细胞分泌凋亡相关的细胞因子。此外，它们还对 NAMALWA 异种移植小鼠模型具有显着的抗肿瘤作用。目前的研究结果为两种扩增方法的安全性和优势提供了证据，这些方法用于转基因 CAR-T 细胞 而与珠子相比，用 aAPC 扩增的 CAR-T 细胞中耗尽分子的表达较低。两种 CAR-T 细胞均显示出显着的靶向杀肿瘤作用。用 aAPC 刺激的 CAR-T 细胞分泌凋亡相关的细胞因子。此外，它们还对 NAMALWA 异种移植小鼠模型具有显着的抗肿瘤作用。目前的研究结果为两种扩增方法的安全性和优势提供了证据，这些方法用于转基因 CAR-T 细胞piggyBac转座子系统。