Division of Immunotherapy, The Hiram C. Polk, Jr., MD Department of Surgery, Immuno-Oncology Program, Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USA; Department of Head and Neck Radiotherapy, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Zhejiang 310022, China.
Division of Immunotherapy, The Hiram C. Polk, Jr., MD Department of Surgery, Immuno-Oncology Program, Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USA.
STAR Protoc. 2023 Sep 15;4(3):102532. doi: 10.1016/j.xpro.2023.102532. Epub 2023 Aug 25.
Intracellular ATP supports the function of γδT17 cells in mice. Here, we present a protocol for intracellular ATP delivery to in vitro expanded mouse CD27 γδ T cells. We describe steps for pre-coating well plates, preparing lymphocytes, culturing CD27 γδ T cells, and ATP delivery. We then detail functional evaluation of γδ T cells by flow cytometry. Appropriate concentrations of control and ATP vesicles are detailed for intracellular ATP delivery, which can also be applied to other immune cells. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023)..
细胞内 ATP 支持小鼠 γδT17 细胞的功能。本文提供了一种将细胞内 ATP 递送至体外扩增的小鼠 CD27 γδ T 细胞的方案。我们描述了包被酶标板、制备淋巴细胞、培养 CD27 γδ T 细胞和 ATP 递送至细胞内的步骤。然后，我们详细介绍了通过流式细胞术评估 γδ T 细胞的功能。我们详细说明了用于细胞内 ATP 递送的对照和 ATP 囊泡的适当浓度，该方法也可应用于其他免疫细胞。有关该方案使用和实施的完整详细信息，请参考 Wang 等人（2023 年）。
