Expanded and Activated Natural Killer Cells for Immunotherapy of Hepatocellular Carcinoma
Expanded and Activated Natural Killer Cells for Immunotherapy of Hepatocellular Carcinoma
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扩增和活化的自然杀伤细胞用于肝细胞癌的免疫治疗
DOI：
10.1158/2326-6066.cir-15-0229
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发表时间：
2016-07-01
影响因子：
10.1
通讯作者：
Campana, Dario
中科院分区：
文献类型：
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作者：
Kamiya, Takahiro;Chang, Yu-Hsiang;Campana, Dario
肝脏病毒感染是肝细胞癌（HCC）的主要危险因素。自然杀伤（NK）细胞识别病毒感染和致癌转化细胞，这表明NK细胞输注在HCC中的治疗作用。以K562-mb 15 - 41 BBL细胞为刺激物，我们从健康供体的外周血中获得了大量活化的NK细胞。扩增的NK细胞对HCC细胞系表现出非常高的细胞毒性，其通常比未刺激的或IL 2激活的NK细胞的细胞毒性高得多。在移植有Hep 3B的免疫缺陷NOD/scid IL 2 RGnull小鼠中，用扩增的NK细胞治疗显著降低了肿瘤生长并改善了总体存活率。HCC细胞暴露于5 mmol/L索拉非尼（目前用于HCC治疗的激酶抑制剂）48小时后，对扩增的NK细胞保持高度敏感。索拉非尼在三种细胞系中引起的HCC细胞减少为39.2%至53.8%，在与NK细胞以1：1的效应细胞与靶细胞比率培养4小时后进一步增加至80.5%至87.6%。即使在索拉非尼存在下，NK细胞的细胞毒性也持续存在。我们发现NKG 2D是一种NK细胞激活受体，是抗HCC活性的重要介质。因此，我们用嵌合NKG 2D-CD 3 zeta-DAP 10受体增强了其信号传导能力。这大大增加了体外和免疫缺陷小鼠中扩增的NK细胞的抗HCC细胞毒性。本研究中应用的NK扩增和活化方法已适应临床级条件。因此，这些结果保证了在HCC患者中进行扩大NK细胞输注的临床试验，可能是在用NKG 2D-CD 3 zeta-DAP 10进行遗传修饰后。
Viral infection of the liver is a major risk factor for hepatocellular carcinoma (HCC). Natural killer (NK) cells recognize virally infected and oncogenically transformed cells, suggesting a therapeutic role for NK-cell infusions in HCC. Using the K562-mb15-41BBL cell line as a stimulus, we obtained large numbers of activated NK cells from the peripheral blood of healthy donors. Expanded NK cells exerted remarkably high cytotoxicity against HCC cell lines, which was generally much higher than that of unstimulated or IL2-activated NK cells. In immunodeficient NOD/scid IL2RGnull mice engrafted with Hep3B, treatment with expanded NK cells markedly reduced tumor growth and improved overall survival. HCC cells exposed for 48 hours to 5 mmol/L of sorafenib, a kinase inhibitor currently used for HCC treatment, remained highly sensitive to expanded NK cells. HCC cell reductions of 39.2% to 53.8% caused by sorafenib in three cell lines further increased to 80.5% to 87.6% after 4 hours of culture with NK cells at a 1: 1 effector-to-target ratio. NK-cell cytotoxicity persisted even in the presence of sorafenib. We found that NKG2D, an NK-cell-activating receptor, was an important mediator of anti-HCC activity. We therefore enhanced its signaling capacity with a chimeric NKG2D-CD3 zeta-DAP10 receptor. This considerably increased the anti-HCC cytotoxicity of expanded NK cells in vitro and in immunodeficient mice. The NK expansion and activation method applied in this study has been adapted to clinical-grade conditions. Hence, these results warrant clinical testing of expanded NK-cell infusions in patients with HCC, possibly after genetic modification with NKG2D-CD3 zeta-DAP10.