Department of Immunology, Leibniz Research Centre for Working Environment and Human Factors at TU Dortmund (IfADo), Dortmund, Germany.
Department of Immunology, Leibniz Research Centre for Working Environment and Human Factors at TU Dortmund (IfADo), Dortmund, Germany.
Methods Cell Biol. 2023;178:13-24. doi: 10.1016/bs.mcb.2022.09.022. Epub 2022 Nov 3.
Natural Killer (NK) cells are innate lymphocytes that are important for early immune reactions against viral infections and cancer. Their cytotoxic activity is mediated by the release of perforin and granzymes or by engaging death receptors on the surface of their target cells. Here we provide a protocol for the use of fluorescence localization reporters to measure the activity of granzyme B or caspase-8 activity inside living target cells. This method can be used to investigate how these two killing pathways are used by NK cells. By modifying the modular structure of the reporters, they can be adapted to study other cytotoxic effector cells or signaling pathways, where proteases play an important role.
自然杀伤 (NK) 细胞是先天淋巴细胞，对于针对病毒感染和癌症的早期免疫反应很重要。它们的细胞毒性活性是通过释放穿孔素和颗粒酶或通过与靶细胞表面的死亡受体结合来介导的。在这里，我们提供了一种使用荧光定位报告分子来测量活靶细胞内颗粒酶 B 或半胱天冬酶-8 活性的方案。该方法可用于研究 NK 细胞如何使用这两种杀伤途径。通过修饰报告分子的模块化结构，可以将其用于研究其他细胞毒性效应细胞或信号通路，其中蛋白酶发挥着重要作用。
