Educated natural killer (NK) cells have inhibitory receptors specific for self major histocompatibility complex (MHC) class I molecules and kill cancer cells more efficiently than do NK cells that do not have such receptors (hyporesponsive NK cells). The mechanism behind this functional empowerment through education has so far not been fully described. In addition, distinctive phenotypic markers of educated NK cells at the single-cell level are lacking. We developed a refined version of the image mean square displacement (iMSD) method (called iMSD carpet analysis) and used it in combination with single-particle tracking to characterize the dynamics of the activating receptor NKp46 and the inhibitory receptor Ly49A on resting educated versus hyporesponsive murine NK cells. Most of the NKp46 and Ly49A molecules were restricted to microdomains; however, individual NKp46 molecules resided in these domains for shorter periods and diffused faster on the surface of educated, compared to hyporesponsive, NK cells. In contrast, the movement of Ly49A was more constrained in educated NK cells compared to hyporesponsive NK cells. Either disrupting the actin cytoskeleton or adding cholesterol to the cells prohibited activating signaling, suggesting that the dynamics of receptor movements within the cell membrane are critical for the proper activation of NK cells. The faster and more dynamic movement of NKp46 in educated NK cells may facilitate a swifter response to interactions with target cells.

受过教育的自然杀伤（NK）细胞具有对自身主要组织相容性复合物（MHC）I类分子特异的抑制受体，比不具有此类受体的NK细胞（低反应性NK细胞）更有效地杀死癌细胞。迄今为止，尚未充分描述这种通过教育赋予职​​能的背后机制。此外，缺乏在单细胞水平上受过训练的NK细胞的独特表型标记。我们开发了图像均方位移（iMSD）方法的改进版本（称为iMSD地毯分析），并将其与单粒子跟踪结合使用，以表征激活受体NKp46和抑制性受体Ly49A在静息受教育与低反应之间的动力学关系鼠NK细胞。大部分的NKp46和Ly49A分子都被限制在微结构域中。但是，与反应低下的NK细胞相比，单个NKp46分子在这些域中的驻留时间更短，并且在受过教育的表面扩散得更快。相反，与低反应性NK细胞相比，在受过教育的NK细胞中Ly49A的运动更受限制。破坏肌动蛋白的细胞骨架或向细胞中添加胆固醇均会阻止激活信号传导，这表明细胞膜内受体运动的动力学对于正确激活NK细胞至关重要。NKp46在受过训练的NK细胞中更快，更动态地运动可能有助于更快地响应与靶细胞的相互作用。与低反应性NK细胞相比，受过教育的NK细胞中Ly49A的运动受到更多的限制。破坏肌动蛋白的细胞骨架或向细胞中添加胆固醇均会阻止激活信号传导，这表明细胞膜内受体运动的动力学对于正确激活NK细胞至关重要。NKp46在受过训练的NK细胞中更快，更动态地运动可能有助于更快地响应与靶细胞的相互作用。与低反应性NK细胞相比，受过教育的NK细胞中Ly49A的运动受到更多的限制。破坏肌动蛋白的细胞骨架或向细胞中添加胆固醇均会阻止激活信号传导，这表明细胞膜内受体运动的动力学对于正确激活NK细胞至关重要。NKp46在受过训练的NK细胞中更快，更动态地运动可能有助于更快地响应与靶细胞的相互作用。