上海交通大学学报(医学版) ›› 2020, Vol. 40 ›› Issue (11): 1437-1446.doi: 10.3969/j.issn.1674-8115.2020.11.001

• 创新团队成果专栏 • 上一篇    下一篇

STAT3对免疫缺陷鼠过继转移CD8+ T细胞的调节作用

曾群雄1, 2,邓 军1, 2,沈 南1, 2   

  1. 1. 上海交通大学医学院附属仁济医院,上海市风湿病学研究所,上海200127;2. 上海交通大学医学院附属仁济医院中澳个体化自身免疫病研究中心,上海200127
  • 出版日期:2020-11-28 发布日期:2021-01-13
  • 通讯作者: 沈 南,电子信箱:nanshensibs@gmail.com。
  • 作者简介:曾群雄(1992—),男,硕士生;电子信箱:zengqunxiong@icloud.com。
  • 基金资助:
    国家自然科学基金(31600708,81974252,81421001,81571575,81771737);上海交通大学医学院高水平地方高校创新团队(SSMU-ZDCX20180100)。

Study the regulatory role of STAT3 on CD8+ T cells using adoptive transfer model in immunodeficient mice

ZENG Qun-xiong1, 2, DENG Jun1, 2, SHEN Nan1, 2   

  1. 1. Shanghai Institute of Rheumatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China; 2. China-Australia Centre for Personalized Immunology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Online:2020-11-28 Published:2021-01-13
  • Supported by:
    National Natural Science Foundation of China (31600708, 81974252, 81421001, 81571575, 81771737); Innovative Research Team of High-Level Local Universities in Shanghai (SSMU-ZDCX20180100).

摘要: 目的·探讨转录因子STAT3对 CD8+ T细胞在免疫缺陷Rag1-/-小鼠中的增殖及活化的内在调控作用。方法·流式分选CD45.1小鼠和CD45.2-CD8(ΔStat3)小鼠的Na?ve CD8+ T细胞,等数量混合并用羟基荧光素二醋酸盐琥珀酰亚胺酯(carboxyfluorescein diacetate succinimidyl ester,CFSE)标记后尾静脉过继转移到Rag1-/-小鼠。细胞转移10 d后,分离脾脏和淋巴结并收集细胞,用流式细胞术检测脾脏、淋巴结中的CD8+ T细胞的比率、增殖情况;用佛波酯(phorbol myristate acetate,PMA)和离子霉素(ionomycin,Ion)刺激后检测细胞因子和杀伤性分子的表达情况;运用t检验分析Stat3敲除后对CD8+ T细胞活化和效应分子表达的影响。结果·在脾脏、腹股沟和肠系膜淋巴结中,野生型(wild type,WT) CD8+ T细胞的比例显著高于ΔStat3 CD8+ T细胞(76.2% vs 23.4%,82.1% vs 17.4%,64.5% vs 32.3%,均P=0.008);腹股沟淋巴结中WT CD8+ T细胞活化的CD44+T细胞比例和细胞数量显著高于ΔStat3 CD8+ T细胞(均P=0.008)。脾脏、腹股沟淋巴结和肠系膜淋巴结中,WT CD8+ T细胞的TNF-α、IFN-γ、GraB、CD107a的表达水平显著高于ΔStat3 CD8+ T细胞(均P<0.05)。结论·Rag1-/-小鼠过继转移Na?ve CD8+ T细胞是研究信号通路/转录因子调控CD8+ T细胞功能的理想体内实验模型,STAT3可内在调控CD8+ T细胞的活化、增殖和效应功能。

关键词: CD8+ T细胞, 过继转移, STAT3, 存活, 功能

Abstract:

Objective · To investigate the intrinsically regulatory role of transcription factor STAT3 on the activation and proliferation of CD8+ T cells in immunodeficient Rag1-/- mice. Methods · The CD8+CD44-CD62L+Na?ve T cells, from CD45.1 wild type (WT) mice and CD45.2-CD8 (ΔStat3) mice, were sorted by flow cytometry cell sorter, equally mixed and labelled with Carboxyl Fluorescein diacetate Succinimidyl Ester (CFSE), and then transferred (i.v.) to the Rag1-/- mice. Ten days after CD8+ T cells transfer, the proportion and proliferation of CD8+ T cells in the spleen, mesenteric lymph nodes (mLNs) and popliteal lymph nodes (pLNs) were determined by flow cytometry. TNF-α, IFN-γ, FasL and granzyme B (GraB) produced by the CD8+ T cells were measured with flow cytometry after PMA and ionomycin stimulation. The statistical significance of activation and effectors expressed between WT and ΔStat3 CD8+ T cell was analyzed by t test. Results · The percentages of WT CD8+ T cells in spleen, mLNs and pLNs were significantly higher than that in ΔStat3 CD8+ T cells (76.2% vs 23.4%, 82.1% vs 17.4%, 64.5% vs 32.3%, with all P=0.008). The percentage and cell number of activated CD44+CD8+ T cells in WT CD8+ T cells were much higher than those in ΔStat3 CD8+ T cells in pLNs (all P=0.008). In spleen, pLNs and mLNs, the levels of TNF-α, IFN-γ, GraB and CD107a expressed in WT CD8+ T cells were higher than those in ΔStat3 CD8+ T cells (all P<0.05). Conclusion · Rag1-/- mice can work as an ideal model to evaluate the survival, proliferation, activation and function of CD8+ T cells in vivo. STAT3 intrinsically regulates the proliferation, activation and function of CD8+ T cells in vivo.

Key words: CD8+ T cell, adoptive transfer, STAT3, survival, function

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