Notch信号通路对慢性丙型肝炎患者CD8<sup>+</sup>T淋巴细胞功能的影响

李彧; 张鸿; 李竹; 刘清娥; 王璐; 张丹; 曹宁家; 苏莉

doi:10.3969/j.issn.1001-5256.2018.02.014

Notch信号通路对慢性丙型肝炎患者CD8⁺T淋巴细胞功能的影响

DOI: 10.3969/j.issn.1001-5256.2018.02.014

1. 陕西省人民医院感染性疾病科
2. 兰州大学公共卫生学院

基金项目:

陕西省科学技术研究发展计划项目(2012SF2-01)；

详细信息

中图分类号: R512.63
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出版历程
- 出版日期: 2018-02-20

Influence of the Notch signaling pathway on CD8⁺T lymphocyte function in patients with chronic hepatitis C

Department of Infectious Diseases, Shaanxi Provincial People′s Hospital

Research funding:

摘要

摘要:
目的观察Notch信号通路对慢性丙型肝炎患者CD8+T淋巴细胞功能的影响,初步阐释Notch信号通路在慢性丙型肝炎发病中的作用。方法收集2016年1月-8月在陕西省人民医院就诊的慢性丙型肝炎初治患者38例和健康志愿者17例,分选CD8+T淋巴细胞,应用实时定量PCR法检测CD8+T淋巴细胞中Notch1和Notch2 mRNA的表达水平。分别应用直接接触培养系统和间接接触培养系统将从HLA-A2限制性丙型肝炎患者分选的CD8+T淋巴细胞与HCV感染的Huh7.5细胞共培养,加入Notch信号通路抑制剂DAPT,通过检测乳酸脱氢酶和细胞因子水平分析CD8+T淋巴细胞的细胞毒性和非细胞毒性功能的变化。2组间计量资料的比较采用t检验。结果 Notch1和Notch2 mRNA在慢性丙型肝炎患者CD8+T淋巴细胞中的表达水平较健康志愿者均显著升高,分别升高约10倍和4倍,差异均有统计学意义（t值分别为6.63、18.88,P值分别为0.003、<0.001）...
- 肝炎,丙型,慢性 /
- CD8阳性T淋巴细胞 /
- 信号传导
Abstract:
Objective To investigate the influence of the Notch signaling pathway on CD8+T lymphocyte function in patients with chronic hepatitis C (CHC) and the role of the Notch signaling pathway in the pathogenesis of CHC.Methods A total of 38 treatment-na6 ve CHC patients who visited Shaanxi Provincial People's Hospital from January to August, 2016 and 17 healthy volunteers were enrolled in this study.CD8+T lymphocytes were isolated, and quantitative real-time PCR was used to measure the mRNA expression of Notch1 and Notch2 in CD8+T lymphocytes.CD8+T lymphocytes, which were isolated from HLA-A2-restricted CHC patients, were co-cultured with hepatitis C virus (HCV) -infected Huh7.5 cells in both direct and indirect contact culture systems.DAPT, a Notch signaling pathway inhibitor, was added, and the changes in cytotoxic and non-cytotoxic functions of CD8+T lymphocytes were analyzed by measuring the levels of lactate dehydrogenase level and cytokines.The t-test was used for comparison of continuous data between groups.Results The mRNA expression of Notch1 and Notch2 in CD8+T lymphocytes in CHC patients was 10 and 4 times, respectively, that in healthy volunteers (t = 6.63, P = 0.003; t = 18.88, P < 0.001) .Inhibition of the Notch signaling pathway significantly increased the percentage of dead cells in the direct contact culture system (26.41 ± 4.76% vs 13.65 ± 4.65%, t = 3.32, P = 0.029) , while it did not influence the cytotoxity of CD8+T lymphocytes in the indirect contact culture system.After inhibition of the Notch signaling pathway, there was a significant increase in the expression of interferon-γ in the direct (52.19 ± 12.38 pg/ml vs 18.62 ± 5.66 pg/ml, t = 4.27, P = 0.013) and indirect contact culture systems (18.86 ± 3.09 pg/ml vs 5.52 ± 2.52 pg/ml, t = 5.79, P = 0.004) .After inhibition of the Notch signaling pathway, there was a significant increase in the expression of tumor necrosis factor-α in the direct contact culture system (1296.0 ± 293.3 pg/ml vs 585.3 ± 32.62 pg/ml, t = 4.17, P = 0.014) , while there was no significant change in the indirect contact culture system (297.3 ± 175.4 pg/ml vs 273.7 ± 105.9 pg/ml, t = 0.20, P = 0.851) .Conclusion Inhibition of the Notch signaling pathway can enhance the cytotoxic and non-cytotoxic functions of CD8+T lymphocytes in CHC patients.The Notch signaling pathway may induce immune tolerance in chronic HCV infection by inhibiting the function of CD8+T lymphocytes.
- hepatitis C, chronic /
- CD8-positive T-lymphocytes /
- signal transduction

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