Real-time Monitoring of Mitochondrial Respiration in Cytokine-differentiated Human Primary T Cells

Kasper M&#248;lgaard; Anne Rahbech; &#214;zcan Met; Inge Marie Svane; Per thor Straten; Claus Desler; Marlies J. W. Peeters

doi:10.3791/62984

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Real-time Monitoring of Mitochondrial Respiration in Cytokine-differentiated Human Primary T Cells

实时监测细胞因子分化人原代T细胞中的线粒体呼吸

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06:55 min

October 19, 2021

DOI: 10.3791/62984-v

Kasper Mølgaard*¹, Anne Rahbech*¹, Özcan Met¹, Inge Marie Svane¹, Per thor Straten^1,3, Claus Desler², Marlies J. W. Peeters¹

¹National Center for Cancer Immune Therapy, Department of Oncology,University Hospital Herlev, ²Department of Cellular and Molecular Medicine, Center for Healthy Aging,University of Copenhagen, ³Department of Immunology and Microbiology, Inflammation and Cancer Group,University of Copenhagen

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代谢适应是 T 细胞的基础，因为它决定了分化、持久性和细胞毒性。本文介绍了一种用于监测离体细胞因子分化的人原代T细胞中线粒体呼吸的优化方案。

线粒体代谢是T细胞耐久性和记忆力的重要调节剂。因此，测量线粒体呼吸可以对T细胞的性质提供重要的见解。海马机可以实时测量活人原发性T淋巴细胞中线粒体呼吸的各种元素，而无需任何标记。

该方法非常适合监测T细胞的适应性和记忆潜力，是癌症免疫治疗成功的重要预测因素。首先，通过将每百万个细胞中12.5微升微粒转移到微量离心管中，并在管中每12.5微升微升珠子中加入12.5微升PBS来洗涤CD3 / CD28微珠。然后，将微量离心管放在合适的磁铁上一分钟，丢弃缓冲液，并将微珠重悬于原始体积的T细胞培养基中。

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