Expansion of highly cytotoxic human natural killer cells for cancer cell therapy.
Expansion of highly cytotoxic human natural killer cells for cancer cell therapy.
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DOI：
10.1158/0008-5472.can-08-3712
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发表时间：
2009-05-01
期刊：
影响因子：
11.2
通讯作者：
Campana D
中科院分区：
文献类型：
--
作者：
Fujisaki H;Kakuda H;Shimasaki N;Imai C;Ma J;Lockey T;Eldridge P;Leung WH;Campana D
自然杀伤（NK）细胞的输注是癌症免疫治疗的新兴工具。开发临床适用的方法来产生大量功能齐全的NK细胞是最大限度地发挥这种方法潜力的关键步骤。我们测定了经修饰以表达膜结合形式的白细胞介素-15和4-1BB配体（K562-mb 15 - 4 - 1BBL）的白血病细胞系K562产生具有增强的细胞毒性的人NK细胞的能力。与经辐照的K562-mb 15 - 41 BBL共培养7天，诱导外周血中CD 56 + CD 3- NK细胞的中值扩增为21.6倍（范围，5.1-86.6倍; n = 50），这显著上级用白细胞介素（IL）-2，IL-12，IL-15和/或IL-21刺激产生的扩增，并且不引起CD 3+淋巴细胞的增殖。从接受治疗的急性白血病患者（n = 11）的外周血中也可以获得类似的扩增。扩增的NK细胞的基因表达谱和它们的未刺激的或IL-2刺激的对应物的比较表现出显着的差异。扩增的NK细胞在体外对急性髓性白血病（AML）细胞的杀伤力明显高于未刺激或IL-2刺激的NK细胞。当注射到免疫缺陷小鼠中时，它们可以被检测到超过一个月，并且可以根除AML小鼠模型中的白血病。因此，我们将K562-mb 15 - 41 BBL刺激方法适应于大规模临床级条件，产生大量高细胞毒性NK细胞。本研究结果为扩增和活化的NK细胞用于肿瘤细胞治疗的临床试验提供了理论依据和实践平台。
Infusions of natural killer (NK) cells are an emerging tool for cancer immunotherapy. The development of clinically applicable methods to produce large numbers of fully functional NK cells is a critical step to maximize the potential of this approach. We determined the capacity of the leukemia cell line K562 modified to express a membrane-bound form of interleukin-15 and 4-1BB ligand (K562-mb15-41BBL) to generate human NK cells with enhanced cytotoxicity. Seven-day coculture with irradiated K562-mb15-41BBL induced a median 21.6-fold expansion of CD56+CD3- NK cells from peripheral blood (range, 5.1-86.6-fold; n = 50), which was considerably superior to that produced by stimulation with interleukin (IL)-2, IL-12, IL-15 and/or IL-21 and caused no proliferation of CD3+ lymphocytes. Similar expansions could also be obtained from the peripheral blood of patients with acute leukemia undergoing therapy (n = 11). Comparisons of the gene expression profiles of the expanded NK cells and of their unstimulated or IL-2-stimulated counterparts demonstrated marked differences. The expanded NK cells were significantly more potent than unstimulated or IL-2-stimulated NK cells against acute myeloid leukemia (AML) cells in vitro. They could be detected for more than one month when injected into immunodeficient mice and could eradicate leukemia in murine models of AML. We therefore adapted the K562-mb15-41BBL stimulation method to large-scale clinical-grade conditions, generating large numbers of highly cytotoxic NK cells. The results that we report here provide rationale and practical platform for clinical testing of expanded and activated NK cells for cell therapy of cancer.