Center for Pediatric Cancer, Department of Pediatrics, National Cancer Center, Goyang 10408, Korea.
Department of Medical Biotechnology, College of Biomedical Science, Kangwon National University, Chuncheon 24341, Korea.
Cells. 2022 Jul 11;11(14):2164. doi: 10.3390/cells11142164.
Immunotherapy is an attractive therapeutic strategy for the treatment of osteosarcoma (OS). The unique features of γδ T cells have made them popular for cancer immunotherapy. Here, we expanded γδ T cells using human peripheral blood mononuclear cells (PBMCs) and investigated their therapeutic potential against OS cells. PBMCs from healthy donors were cultured for 10 days with CON medium (unstimulated control); EX media, CON with recombinant human interleukin-2 (rhIL-2) and zoledronate; and EX28 media, CON with rhIL-2, zoledronate, and CD3/CD28 activator. The expanded γδ T cells were isolated by magnetic cell separation or fluorescence-activated cell sorting, cultured with two OS cell lines (KHOS/NP and MG-63) at various cell ratios with or without doxorubicin or ifosfamide, and analyzed for cytotoxicity and cytokine secretion. The number of CD3γδTCRVγ9 triple-positive γδ T cells and concentrations of IFN-γ and TNF-α were highest in the rhIL-2 (100 IU) and zoledronate (1 μM) supplemented culture conditions. The CD3/CD28 agonist did not show any additional effects on γδ T cell expansion. The expanded γδ T cells exhibited potent in vitro cytotoxicity against OS in a ratio- and time-dependent manner. The γδ T cells may enhance the effect of chemotherapeutic agents against OS and may be a new treatment strategy, including chemo-immunotherapy, for OS.
免疫疗法是治疗骨肉瘤（OS）的一种有吸引力的治疗策略。γδ T 细胞的独特特征使其成为癌症免疫治疗的热门选择。在这里，我们使用人外周血单核细胞（PBMC）扩增 γδ T 细胞，并研究其对 OS 细胞的治疗潜力。健康供体的 PBMC 在 CON 培养基（未刺激对照）中培养 10 天；EX 培养基，CON 加重组人白细胞介素 2（rhIL-2）和唑来膦酸；和 EX28 培养基，CON 加 rhIL-2、唑来膦酸和 CD3/CD28 激活剂。通过磁性细胞分离或荧光激活细胞分选分离扩增的 γδ T 细胞，与两种 OS 细胞系（KHOS/NP 和 MG-63）在有或没有多柔比星或异环磷酰胺的不同细胞比例下培养，并分析细胞毒性和细胞因子分泌。在 rhIL-2（100IU）和唑来膦酸（1μM）补充培养条件下，CD3γδTCRVγ9 三阳性 γδ T 细胞的数量和 IFN-γ 和 TNF-α 的浓度最高。CD3/CD28 激动剂对 γδ T 细胞扩增没有显示出任何额外的作用。扩增的 γδ T 细胞以比例和时间依赖的方式对 OS 表现出强大的体外细胞毒性。γδ T 细胞可能增强化疗药物对 OS 的作用，并且可能是一种新的治疗策略，包括化疗免疫治疗，用于 OS。
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