Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Department of Research Affairs, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Cannabis Cannabinoid Res. 2024 Jun;9(3):819-829. doi: 10.1089/can.2023.0108. Epub 2023 Oct 25.
CD19-chimeric antigen receptor (CAR) T cell therapy is a promising immunotherapy for cancer treatment that has shown remarkable clinical responses, leading to approval by the FDA for relapsed and refractory B cell hematological malignancy treatment. Cannabidiol (CBD) is a nonpsychoactive cannabinoid compound that has been utilized as a palliative treatment in cancer patients due to its immunosuppressive properties. Currently, studies on using CBD during immunotherapy have gained increasing attention. However, the possible interaction between CBD and CAR T cell therapy has not been studied. Therefore, in this study, we aimed to examine the direct effects of CBD on CD19-CAR T cell function against hematologic malignancies. The cytotoxic effect of CBD was determined by a cell proliferation reagent water-soluble tatrazolium salt (WST-1) assay. CAR T cells were generated by retroviral transduction and treated with CBD at a nontoxic dose. The effect of CBD on immune characteristics, including transgene expression, T cell subset, and memory phenotype, was analyzed by flow cytometry. Proliferation, apoptosis, and cell cycle distribution were analyzed with standard methods. The effect on cytotoxic function was evaluated using degranulation assays, and antitumor activity was evaluated using flow cytometry. The half-maximum inhibitory concentration (IC) of CBD on NALM6, Raji, and T cells ranged from 16 to 22 μM. The maximum nontoxic dose of CBD that maintained cell viability at ∼100% was 8 μM. For the generation of CD19-CAR T cells, primary T cells were activated and transduced with a retroviral vector encoding CD19-CAR. CBD did not alter the surface expression or immune characteristics, including the T cell subset and memory phenotype, of CD19-CAR T cells. However, CBD suppressed CD19-CAR T cell proliferation by inducing apoptosis, as evidenced by an increase in the proportion of cells in the Sub-G1 phase in cell cycle arrest. However, the antitumor activity and cytokine secretion of CD19-CAR T cells were not altered by exposure to CBD in this study. In this study, a nontoxic dose of CBD affected CD19-CAR T cell proliferation but not its immune characteristics or cytotoxic function.
CD19 嵌合抗原受体 (CAR) T 细胞疗法是一种有前途的癌症治疗免疫疗法，已显示出显著的临床反应，因此已被 FDA 批准用于治疗复发和难治性 B 细胞血液恶性肿瘤。大麻二酚 (CBD) 是一种非精神活性大麻素化合物，由于其具有免疫抑制特性，已被用作癌症患者的姑息治疗药物。目前，关于在免疫治疗期间使用 CBD 的研究受到了越来越多的关注。然而，CBD 与 CAR T 细胞疗法之间可能存在相互作用尚未得到研究。因此，在这项研究中，我们旨在研究 CBD 对针对血液恶性肿瘤的 CD19-CAR T 细胞功能的直接影响。通过细胞增殖试剂水溶性四唑盐 (WST-1) 测定来确定 CBD 的细胞毒性作用。通过逆转录病毒转导生成 CAR T 细胞，并在无毒剂量下用 CBD 处理。通过流式细胞术分析 CBD 对免疫特征的影响，包括转基因表达、T 细胞亚群和记忆表型。通过标准方法分析增殖、凋亡和细胞周期分布。通过脱颗粒测定评估对细胞毒性功能的影响，并通过流式细胞术评估抗肿瘤活性。CBD 对 NALM6、Raji 和 T 细胞的半最大抑制浓度 (IC) 范围为 16 至 22 μM。维持细胞活力约 100%的 CBD 的最大无毒剂量为 8 μM。为了生成 CD19-CAR T 细胞，将原代 T 细胞激活并转导编码 CD19-CAR 的逆转录病毒载体。CBD 不会改变 CD19-CAR T 细胞的表面表达或免疫特征，包括 T 细胞亚群和记忆表型。然而，CBD 通过诱导细胞凋亡抑制 CD19-CAR T 细胞增殖，这表现为细胞周期阻滞中 Sub-G1 期细胞比例增加。然而，在本研究中，CBD 暴露并未改变 CD19-CAR T 细胞的抗肿瘤活性和细胞因子分泌。在这项研究中，无毒剂量的 CBD 影响 CD19-CAR T 细胞的增殖，但不影响其免疫特征或细胞毒性功能。
