Cytotoxicity of activated natural killer cells against pediatric solid tumors.
Cytotoxicity of activated natural killer cells against pediatric solid tumors.
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DOI：
10.1158/1078-0432.ccr-10-0735
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发表时间：
2010-08-01
期刊：
影响因子：
--
通讯作者：
Campana D
中科院分区：
文献类型：
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作者：
Cho D;Shook DR;Shimasaki N;Chang YH;Fujisaki H;Campana D
为了开发儿童实体瘤的新疗法，我们测试了通过与K562-mb 15 - 41 BBL细胞共培养扩增的自然杀伤（NK）细胞的细胞毒性。我们试图确定最敏感的肿瘤亚型，阐明调节细胞毒性的分子相互作用，并确定体内NK抗肿瘤潜力。我们测试了扩增的NK细胞对代表尤文肉瘤（EWS）（n=5）、横纹肌肉瘤（n=4）、神经母细胞瘤（n=3）和骨肉瘤（n=3）的细胞系的体外细胞毒性，并将结果与抑制性和活化性NK受体配体的表达相关联。我们还比较了扩增和原代NK细胞，确定了激活受体连接和化疗药物的作用，并评估了NK细胞输注在异种移植物中的治疗效果。在45项实验中，EWS和横纹肌肉瘤细胞系对扩增的NK细胞非常敏感，效应细胞：靶细胞比例为1：1时的中位细胞毒性分别为87.2%和79.1%。细胞毒性与NK受体配体的表达水平无关，也不受柔红霉素或长春新碱预处理靶细胞的影响，但NK细胞与NK活化受体NKGD 2和DNAM-1抗体的组合预孵育可显著抑制细胞毒性。扩增的NK细胞比未刺激的NK细胞的细胞毒性大得多，并且根除了移植在NOD/scid IL 2 RGnull小鼠中的EWS细胞。在儿科实体瘤中，EWS和横纹肌肉瘤对扩增的NK细胞非常敏感。本文所述的NK扩增方法已适应于大规模条件，并支持包括这些恶性肿瘤患者的I期临床研究。
To develop new therapies for children with solid tumors, we tested the cytotoxicity of natural killer (NK) cells expanded by coculture with K562-mb15-41BBL cells. We sought to identify the most sensitive tumor subtypes, clarify the molecular interactions regulating cytotoxicity, and determine NK anti-tumor potential in vivo. We tested in vitro cytotoxicity of expanded NK cells against cell lines representative of Ewing sarcoma (EWS) (n=5), rhabdomyosarcoma (n=4), neuroblastoma (n=3) and osteosarcoma (n=3), and correlated the results with expression of inhibitory and activating NK receptor ligands. We also compared expanded and primary NK cells, determined the effects of activating-receptor ligation and of chemotherapeutic drugs, and assessed the therapeutic effect of NK cell infusions in xenografts. In 45 experiments, EWS and rhabdomyosarcoma cell lines were remarkably sensitive to expanded NK cells, with median cytotoxicities at 1:1 effector:target ratio of 87.2% and 79.1%, respectively. Cytotoxicity was not related to levels of expression of NK receptor ligands, nor was it affected by pretreatment of target cells with daunorubicin or vincristine, but was markedly inhibited by preincubation of NK cells with a combination of antibodies against the NK activating receptors NKGD2 and DNAM-1. Expanded NK cells were considerably more cytototoxic than unstimulated NK cells, and eradicated EWS cells engrafted in NOD/scid IL2RGnull mice. Among pediatric solid tumors, EWS and rhabdomyosarcoma are exquisitely sensitive to expanded NK cells. The NK expansion method described here has been adapted to large-scale conditions, and supports a Phase I clinical study including patients with these malignancies.