自然杀伤 (NK) 细胞是先天免疫系统的关键组成部分，参与保护宿主免受病毒感染细胞的侵害和肿瘤免疫监视。在体外培养条件下，IL-12/15/18可以诱导NK细胞产生类似记忆的表型。这些细胞因子诱导的记忆样 (CIML) NK 细胞具有免疫治疗所需的特性，包括更长的寿命和增加的细胞毒性。在这项研究中，从健康捐献者的外周血中分离出 NK 细胞，并用 IL-12/15/18 进行刺激诱导记忆样表型或单独使用 IL-15 作为对照。培养 7 天后，进行多参数流式细胞术分析以评估 CIML 和对照 NK 细胞的表型和功能特征。我们的结果显示 CD25、CD69、NKG2D、NKp30、NKp44、NKp46、TACTILE 和 Granzyme 的表达显着升高B CIML NK 细胞与对照 NK 细胞相比。相反，CIML NK 细胞中的 KIR2D 表达显着低于对照 NK 细胞。此外，功能实验表明，CIML NK 细胞针对靶细胞系 K562 表现出增强的脱颗粒能力和增加的细胞内 IFN-γ 产量。有趣的是，CIML NK 细胞的脱颗粒能力与激活受体 NKp46 和 NKp30 以及抑制性受体 TACTILE 的表达呈正相关。 总之，本研究提供了体外扩增的 CIML NK 细胞的深层表型特征。此外，NK 细胞受体与 CIML NK 细胞脱颗粒能力之间的相关性使得能够鉴定出在临床环境中有用的几种生物标志物。版权所有 © 2023 Carreira-Santos、López-Sejas、González-Sánchez、Sánchez-Hernández、Pera 、哈苏内、杜兰、索拉纳、卡萨多和塔拉索纳。

Natural killer (NK) cells are a key component of the innate immune system, involved in defending the host against virus-infected cells and tumor immunosurveillance. Under in vitro culture conditions, IL-12/15/18 can induce a memory-like phenotype in NK cells. These cytokine-induced memory-like (CIML) NK cells possess desirable characteristics for immunotherapies, including a longer lifespan and increased cytotoxicity.In this study, NK cells were isolated from peripheral blood of healthy donors and stimulated with IL-12/15/18 to induce a memory-like phenotype or with IL-15 alone as a control. After seven days of culture, multiparametric flow cytometry analysis was performed to evaluate the phenotypic and functional profiles of CIML and control NK cells.Our results showed a significantly higher expression of CD25, CD69, NKG2D, NKp30, NKp44, NKp46, TACTILE, and Granzyme B in CIML NK cells compared to control NK cells. In contrast, KIR2D expression was significantly lower in CIML NK cells than in control NK cells. Moreover, functional experiments demonstrated that CIML NK cells displayed enhanced degranulation capacity and increased intracellular IFN-γ production against the target cell line K562. Interestingly, the degranulation capacity of CIML NK cells was positively correlated with the expression of the activating receptors NKp46 and NKp30, as well as with the inhibitory receptor TACTILE.In conclusion, this study provides a deep phenotypic characterization of in vitro-expanded CIML NK cells. Moreover, the correlations found between NK cell receptors and degranulation capacity of CIML NK cells allowed the identification of several biomarkers that could be useful in clinical settings.Copyright © 2023 Carreira-Santos, López-Sejas, González-Sánchez, Sánchez-Hernández, Pera, Hassouneh, Durán, Solana, Casado and Tarazona.