Streltsova Maria A, Barsov Eugene, Erokhina Sofia A, Kovalenko Elena I
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, Moscow 117997, Russia.
Cell Design Labs, Inc., Emeryville, CA, USA.
J Immunol Methods. 2017 Nov;450:90-94. doi: 10.1016/j.jim.2017.08.003. Epub 2017 Aug 10.
Natural killer (NK) cells are capable of rapidly recognizing and efficiently killing tumor cells. This makes them a potentially promising agent for cancer immunotherapy. Additional genetic modifications of NK cells may further improve their anti-tumor efficacy. Numerous technical challenges associated with gene delivery into NK cells have significantly tempered this approach. We achieved efficient retroviral vector transduction of primary human NK cells that were stimulated by a combination of IL-2 and engineered K562 cells expressing membrane-bound IL-21. The activated NK cells were in less differentiated state and expressed NK cell activation receptors NKG2D, NKp30, CD16, and were highly HLA-DR-positive. This NK cell population was highly susceptible to the transduction by both GFP- and NGFR-expressing retroviral vectors, with transduction efficiency exceeding 50%. More mature CD57 NK cell population was generally resistant to retroviral vector transduction because of poor response to the stimulation. Our findings may facilitate retroviral vector-mediated genetic engineering of human primary NK cells for future immunotherapies.
自然杀伤（NK）细胞能够快速识别并有效杀伤肿瘤细胞。这使其成为癌症免疫治疗中一种潜在的有前景的药物。对NK细胞进行额外的基因改造可能会进一步提高其抗肿瘤功效。然而，与将基因导入NK细胞相关的众多技术挑战显著限制了这种方法的应用。我们通过白细胞介素-2（IL-2）与表达膜结合型IL-21的工程化K562细胞联合刺激，实现了对原代人NK细胞的高效逆转录病毒载体转导。活化的NK细胞处于较低分化状态，表达NK细胞活化受体NKG2D、NKp30、CD16，且高度表达HLA-DR。这群NK细胞对表达绿色荧光蛋白（GFP）和神经生长因子受体（NGFR）的逆转录病毒载体转导高度敏感，转导效率超过50%。由于对刺激反应不佳，更成熟的CD57 NK细胞群体通常对逆转录病毒载体转导具有抗性。我们的研究结果可能有助于通过逆转录病毒载体介导对人原代NK细胞进行基因工程改造，以用于未来的免疫治疗。
