Departments of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
Methods Mol Biol. 2022;2463:311-328. doi: 10.1007/978-1-0716-2160-8_21.
Autologous T cells expressing chimeric antigen receptor (CAR) have produced a spectacular response in hematological malignancies. This success of cellular therapy has inspired the exploration of the therapeutic potential of other immune cell types. In this regard, natural killer (NK) cells hold great potential as they can identify tumor cells by mechanisms that are different from those used by T cells and have a high cytotoxic capacity. Their capacity to recognize tumors and killing potency can be further enhanced by genetic modification. Our laboratory has developed a clinically adaptable method to manufacture genetically modified NK cells using retroviral vectors in compliance with current good manufacturing practice regulations. Here, we describe relevant technical procedures, including isolation of peripheral blood mononucleated cells from a leukapheresis product, T-cell depletion, stimulation and transduction of NK cells, and preparation of transduced NK cells for infusion.
嵌合抗原受体 (CAR) 表达的自体 T 细胞在血液系统恶性肿瘤中产生了显著的反应。细胞治疗的成功激发了对其他免疫细胞类型治疗潜力的探索。在这方面，自然杀伤 (NK) 细胞具有很大的潜力，因为它们可以通过不同于 T 细胞的机制识别肿瘤细胞，并且具有高细胞毒性。通过遗传修饰可以进一步增强它们识别肿瘤和杀伤的能力。我们的实验室开发了一种临床可适应的方法，使用逆转录病毒载体制造符合现行良好生产规范法规的基因修饰 NK 细胞。在这里，我们描述了相关的技术程序，包括从白细胞分离产品中分离外周血单个核细胞、T 细胞耗竭、NK 细胞的刺激和转导，以及转导的 NK 细胞输注的准备。
