使用流式细胞术和功能测定分离小鼠衰老相关 CD4 T 细胞的方案。
Protocol for the isolation of mouse senescence-associated CD4 T cells using flow cytometry and functional assays.
机构信息
Department of Regulation of Neurocognitive Disorders (Cyn-K project), Graduate School of Medicine, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan; Department of Immunosenescence, Graduate School of Medicine, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Medical Innovation Center, Graduate School of Medicine, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
出版信息
STAR Protoc. 2023 Sep 15;4(3):102472. doi: 10.1016/j.xpro.2023.102472. Epub 2023 Jul 28.
Senescence-associated (SA) CD4 T cells, which increase with age, may underlie the development of autoimmunity and chronic inflammation, but their pathological function remains understudied. Here, we present a protocol to isolate CD153 SA-T cells and evaluate their characteristic responses upon T cell receptor stimulation. We describe steps for the isolation of CD153 SA-T cells using flow cytometry and in vitro culture with stimulatory antibodies against CD3, CD28, and CD153. We then detail the assessment of the proliferation capacity and cytokine production. For complete details on the use and execution of this protocol, please refer to Fukushima et al. (2022)..
衰老相关（SA）CD4 T 细胞随年龄增长而增加，可能是自身免疫和慢性炎症发展的基础，但它们的病理功能仍研究不足。在这里，我们提供了一种分离 CD153 SA-T 细胞的方案，并评估了它们在 T 细胞受体刺激下的特征性反应。我们描述了使用流式细胞术分离 CD153 SA-T 细胞的步骤，并使用针对 CD3、CD28 和 CD153 的刺激性抗体进行体外培养。然后，我们详细介绍了评估增殖能力和细胞因子产生的方法。有关此方案的使用和执行的完整详细信息，请参阅 Fukushima 等人。（2022）。
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