Leibniz Research Centre for Working Environment and Human Factors, IfADo, TU-Dortmund, D-44139, Dortmund, Germany.
Sci Rep. 2018 Mar 21;8(1):4938. doi: 10.1038/s41598-018-23368-5.
The use of impedance-based label free cell analysis is increasingly popular and has many different applications. Here, we report that a real-time cell analyzer (RTCA) can be used to study the stimulation of Natural Killer (NK) cells. Engagement of NK cells via plate-bound antibodies directed against different activating surface receptors could be measured in real time using the label-free detection of impedance. The change in impedance was dependent on early signal transduction events in the NK cells as it was blocked by inhibitors of Src-family kinases and by inhibiting actin polymerization. While CD16 was the only receptor that could induce a strong change in impedance in primary NK cells, several activating receptors induced changes in impedance in expanded NK cells. Using PBMCs we could detect T cell receptor-mediated T cell activation and CD16-mediated NK cell activation in the same sample. Performing a dose-response analysis for the Src-family kinases inhibitor PP1 we show that T cells are more sensitive to inhibition compared to NK cells. Our data demonstrate that the RTCA can be used to detect physiological activation events in NK cells in a label-free and real-time fashion.
基于阻抗的无标记细胞分析的应用越来越普及，具有许多不同的应用。在这里，我们报告说，实时细胞分析仪（RTCA）可用于研究自然杀伤（NK）细胞的刺激。通过针对不同激活表面受体的板结合抗体与 NK 细胞的结合，可以使用无标记的阻抗检测实时测量。阻抗的变化取决于 NK 细胞中的早期信号转导事件，因为它被Src 家族激酶抑制剂和抑制肌动蛋白聚合所阻断。虽然 CD16 是唯一能够在原代 NK 细胞中诱导阻抗强烈变化的受体，但几种激活受体在扩增的 NK 细胞中诱导了阻抗的变化。使用 PBMC，我们可以在同一样品中检测到 T 细胞受体介导的 T 细胞激活和 CD16 介导的 NK 细胞激活。对 Src 家族激酶抑制剂 PP1 进行剂量反应分析，我们表明 T 细胞比 NK 细胞对抑制更敏感。我们的数据表明，RTCA 可用于无标记和实时方式检测 NK 细胞中的生理激活事件。
