Department of Immunology, School of Medicine, University of Connecticut, Farmington, CT 06030, USA.
STAR Protoc. 2022 Jun 3;3(2):101441. doi: 10.1016/j.xpro.2022.101441. eCollection 2022 Jun 17.
Studying the metabolic fitness of T cells is fundamental to understand how immune responses are regulated. Here, we describe a step-by-step protocol optimized to efficiently generate and isolate effector antigen-specific CD8 T cells using costimulation. We also detail steps to evaluate their metabolic activity using Seahorse technology. This protocol can be used to measure the glycolytic potential of effector murine T cells in response to different manipulations, such as infections, adjuvant studies, gene editing, or metabolite supplementation. For complete details on the use and execution of this protocol, please refer to Agliano et al. (2022).
研究 T 细胞的代谢适应性对于了解免疫反应如何调节至关重要。在这里，我们描述了一个经过优化的分步方案，该方案使用共刺激来高效生成和分离效应抗原特异性 CD8 T 细胞。我们还详细介绍了使用 Seahorse 技术评估其代谢活性的步骤。该方案可用于测量效应鼠 T 细胞对不同操作（如感染、佐剂研究、基因编辑或代谢物补充）的糖酵解潜力。有关此方案的使用和执行的完整详细信息，请参阅 Agliano 等人（2022 年）。
