Intracellular ATP delivery to in vitro expanded mouse CD27^- æ T cells

Intracellular ATP supports the function of gamma delta T17 cells in mice. Here, we present a protocol for intracellular ATP delivery to in vitro expanded mouse CD27(-) gamma delta T cells. We describe steps for pre-coating well plates, preparing lymphocytes, culturing CD27(-) gamma delta T cells, and ATP delivery. We then detail functional evaluation of gamma delta 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).(1).