Characterization of genetically altered, interleukin 2-independent natural killer cell lines suitable for adoptive cellular immunotherapy
Characterization of genetically altered, interleukin 2-independent natural killer cell lines suitable for adoptive cellular immunotherapy
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DOI：
10.1089/10430349950018030
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发表时间：
1999-05-20
影响因子：
4.2
通讯作者：
Klingemann, HG
中科院分区：
文献类型：
--
作者：
Tam, YK;Maki, G;Klingemann, HG
NK-92是一种具有高细胞毒性的自然杀伤（NK）肿瘤细胞系，具有使其成为过继细胞免疫治疗的优秀候选者的特性。然而，NK细胞的细胞毒性依赖于白细胞介素2 （IL-2）等细胞因子。虽然NK-92细胞在停用IL-2后仍能维持一段时间的细胞毒性，但临床使用可能需要用完全活化的细胞进行长时间治疗才能有效消除疾病。外源性给药IL-2支持细胞毒性细胞的能力受到相关毒性的限制。因此，我们描述了通过颗粒介导的基因转移转染人IL-2 (IL-2) cDNA的IL-2依赖性NK-92细胞系，以创建两个IL-2独立变体NK-92MI和NK-92 CI，并描述了它们的特性和与亲本细胞的比较。这两种变异均含有、表达和合成hIL-2 cDNA。NK-92MI细胞中IL-2的合成高于NK-92CI细胞，亲本细胞中不表达IL-2。在功能上，这三种细胞系的细胞毒性相似，并且与il -2非依赖性变异共孵育不会影响造血祖细胞。NK-92MI和NK-92 CI细胞比NK-92细胞对辐射更敏感，较低的辐射剂量抑制了NK-92细胞的增殖，与亲本细胞相比，毒性增加，毒性降低。这里提供的数据表明，我们通过颗粒介导的基因转移创造了两个NK-92的il -2独立变体，它们在几乎所有方面都与亲本细胞相同，包括高细胞毒性活性。这些细胞的非病毒转染使它们适合临床应用。这些不依赖于IL-2的细胞应该允许在不需要外源IL-2支持的情况下，与完全活跃的自然杀伤细胞进行长期治疗。
NK-92 is a highly cytotoxic natural killer (NK) tumor cell line that possesses properties that make it an excellent candidate for adoptive cellular immunotherapy. However, the cytotoxicity of NK cells is dependent on cytokines such as interleukin 2 (IL-2). Although NK-92 cells maintain cytotoxicity for a time after withdrawal of IL-2, clinical use will probably require prolonged treatment with fully activated cells to eliminate disease effectively. The ability to support cytotoxic cells with exogenously administered IL-2 is limited by associated toxicity. Therefore, we describe the transfection of the IL-2-dependent NK-92 cell line with human IL-2 (hIL-2) cDNA by particle-mediated gene transfer to create two IL-2-independent variants, NK-92MI and NK-92 CI, and describe their characterization and comparison with parental cells. Both variants were shown to contain, express, and synthesize the hIL-2 cDNA. IL-2 synthesis was higher in NK-92MI cells compared with NK-92CI cells, with no expression in parental cells. Functionally, the cytotoxicity of all three cell lines was similar and coincubation with IL-2-independent variants did not affect hematopoietic progenitor cells. NK-92MI and NK-92 CI cells were more radiosensitive than NK-92 cells, with proliferation inhibited at lower radiation doses and increased morality and decreased cytotoxicity compared with parental cells. Data presented here show that we have created by particle-mediated gene transfer two IL-2-independent variants of NK-92 that are identical to parental cells in virtually all respects, including high cytotoxic activity. The nonviral transfection of these cells makes them suitable for clinical applications. These IL-2-independent cells should allow prolonged treatment with fully active natural killer cells without the need for exogenous IL-2 support.