Protocol for the isolation of mouse senescence- associated CD4+ T cells using flow cytometry and functional assays

被引：0

作者：

Fukushima, Yuji ^{[1
,2
]}

Minato, Nagahiro ^{[3
]}

Hattori, Masakazu ^{[2
,4
]}

机构：

[1] Kyoto Univ, Grad Sch Med, Dept Regulat Neurocognit Disorders, Cyn K Project, 53 Shogoin Kawahara Cho,Sakyo Ku, Kyoto 6068507, Japan

[2] Kyoto Univ, Grad Sch Med, Dept Immunosenescence, 53 Shogoin Kawahara Cho,Sakyo Ku, Kyoto 6068507, Japan

[3] Kyoto Univ, Grad Sch Med, Med Innovat Ctr, 53 Shogoin Kawahara Cho,Sakyo Ku, Kyoto 6068507, Japan

[4] Kyoto Univ, Grad Sch Med, Lab Tumor Tissue Response, 53 Shogoin Kawahara Cho,Sakyo Ku, Kyoto 6068507, Japan

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 03期

关键词：

Cell Culture; Cell Isolation; Flow Cytometry/Mass Cytometry; Immunology; Signal Transduction;

D O I：

10.1016/j.xpro.2023.102472

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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).1

引用

页数：18