Objective: To explore whether peripheral blood CD8(+)T lymphocyte dysfunction is correlated with the programmed death receptor-1 (PD-1) expression in patients with acute-on-chronic liver failure (HBV-ACLF). Methods: Peripheral blood mononuclear cells (PBMC) were collected from patients with HBV-ACLF and healthy controls. CD8(+)T lymphocytes number and PD-1 expression condition in CD8(+)T lymphocytes were detected by flow cytometry. CD8(+)T lymphocytes isolated from peripheral blood of HBV-ACLF patients were further cultured in vitro. One group was added with PD-L1-IgG fusion protein (ACLF+PD-1 group), and the other group was added with IgG fusion protein (ACLF group). Proliferation ability (ki67), cell viability (CD69), and secretion ability of effector cytokines (IL-2, IFN-γ, TNF-α) were analyzed. Results: 30 cases with HBV-ACLF and healthy controls were enrolled. CD8(+)T lymphocytes absolute number was significantly lower in the peripheral blood of patients with ACLF group (333.88 ± 147.74)/μl than healthy controls (872.50 ± 206.64)/μl (P < 0.001). PD-1 expression in peripheral blood CD8(+)T lymphocytes were significantly increased in ACLF group (13.33% ± 2.52%), (P = 0.027) than healthy controls (7.02% ± 2.12%). In in vitro culture, compared with healthy controls, the peripheral blood CD8(+)T lymphocytes cell viability (CD69), proliferation ability (ki67) (all P < 0.001), and the level of cytokine production (IL-2, IFN-γ, TNF-α) (all P < 0.05) were equally weakened in patients with ACLF group. Compared with ACLF group, CD8(+)T cell viability (CD69), proliferation ability (KI67) (all P < 0.05), and the level of cytokine production were weakened in ACLF+PD-1 group (all P < 0.05). Conclusion: HBV-ACLF patients have CD8(+)T lymphocyte dysfunction. Therefore, PD-1 may have correlation in the regulation of CD8(+)T lymphocyte dysfunction in ACLF patients.
目的： 探索程序性死亡分子1（PD-1）是否参与乙型肝炎相关的慢加急性肝衰竭（HBV-ACLF）患者外周血CD8(+)T淋巴细胞功能障碍。 方法： 收集HBV-ACLF患者和健康对照者外周血，获得外周血单个核细胞（PBMC），流式细胞术检测CD8(+)T淋巴细胞的数量及CD8(+)T淋巴细胞PD-1的表达状况。进一步体外培养HBV-ACLF患者的外周血分选的CD8(+)T细胞，一组加入PD-L1-IgG融合蛋白（ACLF+PD-1组），一组加入IgG融合蛋白（ACLF组），分析其增殖能力（ki67）和CD8(+)T细胞活力（CD69）及分泌效应细胞因子[白细胞介素2（IL-2）、干扰素γ（IFN-γ）、肿瘤坏死因子α（TNF-α）]能力。 结果： 入组的30例HBV-ACLF患者和健康对照者，ACLF患者外周血CD8(+)T细胞绝对数（333.88±147.74）个/μl明显低于健康对照者（872.50±206.64）个/μl（P < 0.001）。相对于健康对照者（7.02%±2.12%），ACLF患者外周血的CD8(+)T淋巴细胞PD-1表达明显升高（13.33%±2.52%），P = 0.027，差异有统计学意义。在体外培养，相对于健康对照，ACLF患者外周血CD8(+)T细胞活力（CD69）和增殖能力（ki67）均减弱（P值均< 0.001）；产生IL-2、IFN-γ、TNF-α水平减弱（均P < 0.001）。相对于ACLF组，ACLF+PD-1组CD8(+)T细胞活力（CD69）和增殖能力（ki67）进一步减弱（P值均< 0.05）；产生IL-2、IFN-γ、TNF-α水平进一步降低（均P < 0.05）。 结论： HBV-ACLF患者存在CD8(+)T淋巴细胞功能障碍，PD-1可能参与调控ACLF患者的CD8(+)T淋巴细胞功能障碍。.
Keywords: Acute and chronic liver failure; Immune function; Programmed death molecule 1; T lymphocyte.