Rey Camille, Jones Katherine L, Stacey Kevin B, Evans Alicia, Worboys Jonathan D, Howell Gareth, Sheppard Sam, Davis Daniel M
Faculty of Biology Medicine and Health, Lydia Becker Institute of Immunology and Inflammation, Manchester, United Kingdom.
Department of Life Sciences, Sir Alexander Fleming Building, Imperial College London, South Kensington, London, United Kingdom.
Front Immunol. 2025 Feb 19;16:1526379. doi: 10.3389/fimmu.2025.1526379. eCollection 2025.
Natural Killer (NK) cells are innate immune cells that can directly detect and kill cancer cells. Understanding the molecular determinants regulating human NK cell cytotoxicity could help harness these cells for cancer therapies. To this end, we compared the transcriptome of NK cell clones derived from human peripheral blood, which were strongly or weakly cytotoxic against 721.221 and other target cells. After one month of culture, potent NK cell clones showed a significant upregulation in genes involved in cell cycle progression, suggesting that proliferating NK cells were particularly cytotoxic. Beyond two months of culture, NK cell clones which were strongly cytotoxic varied in their expression of 28 genes, including and ; NK cells with high levels of expression were weakly cytotoxic while high correlated with strong cytotoxicity. Thus, NK cells were cultured and sorted for expression of CD70 and CD8α, and in accordance with the transcriptomic data, CD70 NK cells showed low cytotoxicity against 721.221 and K562 target cells. Cytotoxicity of CD70 NK cells could be enhanced using blocking antibodies against CD70, indicating a direct role for CD70 in mediating low cytotoxicity. Furthermore, time-lapse microscopy of NK cell-target cell interactions revealed that CD8α NK cells have an increased propensity to sequentially engage and kill multiple target cells. Thus, these two markers relate to NK cell populations which are capable of potent killing (CD70) or serial killing (CD8α).
自然杀伤（NK）细胞是先天性免疫细胞，能够直接检测并杀死癌细胞。了解调节人类NK细胞细胞毒性的分子决定因素有助于利用这些细胞进行癌症治疗。为此，我们比较了源自人外周血的NK细胞克隆的转录组，这些克隆对721.221和其他靶细胞具有强或弱的细胞毒性。培养一个月后，高效NK细胞克隆在参与细胞周期进程的基因中表现出显著上调，这表明增殖的NK细胞具有特别的细胞毒性。培养超过两个月后，具有强细胞毒性的NK细胞克隆在28个基因的表达上存在差异，包括 和 ； 表达水平高的NK细胞细胞毒性弱，而高 与强细胞毒性相关。因此，对NK细胞进行培养并根据CD70和CD8α的表达进行分选，根据转录组数据，CD70 NK细胞对721.221和K562靶细胞的细胞毒性较低。使用抗CD70阻断抗体可增强CD70 NK细胞的细胞毒性，这表明CD70在介导低细胞毒性中起直接作用。此外，对NK细胞与靶细胞相互作用的延时显微镜观察显示，CD8α NK细胞依次接触并杀死多个靶细胞的倾向增加。因此，这两个标志物与能够进行高效杀伤（CD70）或连续杀伤（CD8α）的NK细胞群体相关。
