Genetic modification of primary natural killer cells overcomes inhibitory signals and induces specific killing of leukemic cells
Genetic modification of primary natural killer cells overcomes inhibitory signals and induces specific killing of leukemic cells
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DOI：
10.1182/blood-2004-12-4797
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发表时间：
2005-07-01
期刊：
影响因子：
20.3
通讯作者：
Campana, D
中科院分区：
文献类型：
--
作者：
Imai, C;Iwamoto, S;Campana, D
自然杀伤 (NK) 细胞有望提高同种异体造血移植的治疗潜力，但其有效性受到抑制性 HLA 类型的限制。我们试图通过用针对CD19（恶性B细胞广泛表达的分子）的嵌合受体转导CD56（+）CD3（-）NK细胞来克服这种内在的抵抗力。通过用表达 NK 刺激分子 4-1BB 配体和白细胞介素 15 的 K562 细胞培养外周血单核细胞，确保了用于转导的大量 NK 细胞，培养 3 周后，CD56(+)CD3(-) 细胞的中值扩增超过 1000 倍，且没有 T 淋巴细胞扩增。与 CD3 xi 连接的抗 CD19 受体的表达克服了 NK 抵抗，并显着增强了 NK 细胞介导的对白血病细胞的杀伤。通过在嵌合抗CD19-CD3xi受体中添加4-1BB共刺激分子，这一结果得到显着改善；表达该构建体的 NK 细胞产生的细胞毒性均超过信号受体缺乏 4-1BB 的 NK 细胞，即使自然细胞毒性很明显。添加 4-1 BB 还与细胞活化以及干扰素 γ 和粒细胞-巨噬细胞集落刺激因子的产生增加有关。我们的研究结果表明，NK 细胞强制表达信号受体可能会规避抑制信号，从而为增强同种异体干细胞移植的有效性提供了一种新方法。
Natural killer (NK) cells hold promise for improving the therapeutic potential of allogeneic hematopoietic transplantation, but their effectiveness is limited by inhibitory HLA types. We sought to overcome this intrinsic resistance by transducing CD56(+)CD3(-) NK cells with chimeric receptors directed against CD19, a molecule widely expressed by malignant B cells. An abundance of NK cells for transduction was secured by culturing peripheral blood mononuclear cells with K562 cells expressing the NK-stimulatory molecules 4-1BB ligand and interleukin 15, which yielded a median greater than 1000-fold expansion of CD56(+)CD3(-) cells at 3 weeks of culture, without T-lymphocyte expansion. Expression of anti-CD19 receptors linked to CD3 xi overcame NK resistance and markedly enhanced NK-cell-mediated killing of leukemic cells. This result was significantly improved by adding the 4-1 BB costimulatory molecule to the chimeric anti-CD19-CD3 xi receptor; the cytotoxicity produced by NK cells expressing this construct uniformly exceeded that of NK cells whose signaling receptors lacked 4-1BB, even when natural cytotoxicity was apparent. Addition of 4-1 BB was also associated with increased cell activation and production of interferon gamma and granulocyte-macrophage colony-stimulating factor. Our findings indicate that enforced expression of signaling receptors by NK cells might circumvent inhibitory signals, providing a novel means to enhance the effectiveness of allogeneic stem cell transplantation.