University Children's Hospital III, University Children's Hospital III, Department of Hematology/Oncology Department of Hematology/Oncology, Theodor-Stern-Kai 7.
Haematologica. 2010 Sep;95(9):1579-86. doi: 10.3324/haematol.2009.019885. Epub 2010 Apr 7.
Rhabdomyosarcoma is the most common soft tissue sarcoma in childhood and has a poor prognosis. Here we assessed the capability of ex vivo expanded cytokine-induced killer cells to lyse both alveolar and embryonic rhabdomyosarcoma cell lines and investigated the mechanisms involved.
Peripheral blood mononuclear cells from six healthy donors were used to generate and expand cytokine-induced killer cells. The phenotype and composition of these cells were determined by multiparameter flow cytometry, while their cytotoxic effect against rhabdomyosarcoma cells was evaluated by a europium release assay.
Cytokine-induced killer cells efficiently lysed cells from both rhabdomyosarcoma cell lines. Antibody-mediated masking of either NKG2D molecule on cytokine-induced killer cells or its ligands on rhabdomyosarcoma cells (major histocompatibility antigen related chain A and B and UL16 binding protein 2) diminished this effect by 50%, suggesting a major role for the NKG2D molecule in rhabdomyosarcoma cell killing. No effect was observed after blocking CD11a, CD3 or TCRalphabeta molecules on cytokine-induced killer cells or CD1d on rhabdomyosar-coma cells. Remarkably, cytokine-induced killer cells used tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to activate caspase-3, as the main caspase responsible for the execution of apoptosis. Accordingly, blocking TRAIL receptors on embryonic rhabdomyosarcoma cell lines significantly reduced the anti-tumor effect of cytokine-induced killer cells. About 50% of T cells within the cytokine-induced killer population had an effector memory phenotype, 20% had a naïve phenotype and approximately 30% of the cells had a central memory phenotype. In addition, cytokine-induced killer cells expressed low levels of activation-induced markers CD69 and CD137 and demonstrated a low alloreactive potential.
Our data suggest that cytokine-induced killer cells may be used as a novel adoptive immunotherapy for the treatment of patients with rhabdomyosarcoma after allogeneic stem cell transplantation.
横纹肌肉瘤是儿童中最常见的软组织肉瘤，预后不良。在这里，我们评估了体外扩增的细胞因子诱导的杀伤细胞溶解肺泡和胚胎横纹肌肉瘤细胞系的能力，并研究了涉及的机制。
使用来自六位健康供体的外周血单核细胞生成和扩增细胞因子诱导的杀伤细胞。通过多参数流式细胞术确定这些细胞的表型和组成，通过铕释放测定评估其对横纹肌肉瘤细胞的细胞毒性作用。
细胞因子诱导的杀伤细胞有效地溶解了来自两种横纹肌肉瘤细胞系的细胞。在细胞因子诱导的杀伤细胞上中和 NKG2D 分子或在横纹肌肉瘤细胞上中和其配体（主要组织相容性抗原相关链 A 和 B 和 UL16 结合蛋白 2）可使该效应降低 50%，表明 NKG2D 分子在横纹肌肉瘤细胞杀伤中起主要作用。在细胞因子诱导的杀伤细胞上阻断 CD11a、CD3 或 TCRalphabeta 分子或在横纹肌肉瘤细胞上阻断 CD1d 后未观察到效果。值得注意的是，细胞因子诱导的杀伤细胞使用肿瘤坏死因子相关凋亡诱导配体（TRAIL）激活半胱天冬酶-3，作为执行细胞凋亡的主要半胱天冬酶。因此，阻断胚胎横纹肌肉瘤细胞系上的 TRAIL 受体显著降低了细胞因子诱导的杀伤细胞的抗肿瘤作用。细胞因子诱导的杀伤细胞群体中约 50%的 T 细胞具有效应记忆表型，20%具有幼稚表型，约 30%的细胞具有中央记忆表型。此外，细胞因子诱导的杀伤细胞表达低水平的激活诱导标记物 CD69 和 CD137，并表现出低的同种异体反应性潜力。
我们的数据表明，细胞因子诱导的杀伤细胞可作为异体干细胞移植后治疗横纹肌肉瘤患者的新型过继免疫疗法。
