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目的通过检测口腔扁平苔藓(OLP)患者外周血T细胞中程序性死亡因子1(PD-1)及其配体B7同系物1(B7-H1)表达水平的变化,探讨其临床意义,并观察在阻断B7-H1/PD-1信号通道后对T细胞增殖及细胞因子分泌的影响,探讨其与OLP发生发展的关系。方法选择2014年1月至2017年1月300例OLP患者(OLP组)和300例健康成人(对照组)作为研究对象。应用网纹-萎缩-糜烂(RAE)计分系统评估OLP患者的疾病状态。采用密度梯度离心法分离外周血中的单个核细胞,采用流式细胞仪检测T细胞上B7-H1和PD-1的表达水平。酶联免疫吸附法(ELISA)测定血清中γ-干扰素(IFN-γ)、白介素(IL)-2、IL-4、IL-10和可溶性PD-1(s PD-1)水平。贴壁法去除单核细胞后,将OLP患者的淋巴细胞与健康成人淋巴细胞混合培养,分别加入抗B7-H1单克隆抗体(anti-B7-H1m Ab)及抗PD-1单克隆抗体(anti-PD-1m Ab)阻断B7-H1/PD-1信号通道,观察其对T细胞增殖及细胞因子分泌的影响。结果 OLP组患者外周血T细胞上B7-H1及PD-1的表达水平显著高于对照组(P均<0.01)。OLP组患者外周血T细胞上B7-H1的表达水平与RAE计分呈正相关性(r=0.510,P<0.01),而PD-1的表达与RAE计分系统无相关性(r=0.041,P>0.05)。OLP组患者血清中Th1细胞因子IFN-γ、IL-2水平显著高于对照组(P均<0.01);Th2细胞因子IL-4、IL-10水平显著低于对照组(P均<0.01)。OLP组和对照组患者血清中s PD-1水平无统计学差异(P>0.05)。应用anti-B7-H1m Ab及anti-PD-1m Ab阻断B7-H1/PD-1信号通路后,T细胞的增殖程度明显高于阻断前OLP组(P<0.05),IFN-γ和IL-2的分泌较阻断前OLP组也显著增加(P均<0.05,P<0.01),表明B7-H1/PD-1信号途径可抑制OLP T细胞的增殖及IFN-γ和IL-2的分泌。结论 OLP患者外周血T细胞B7-H1/PD-1的异常表达可能通过负性调控OLP患者T细胞增殖及细胞因子分泌影响OLP的发生发展,OLP外周血T细胞上B7-H1的表达水平可用来监测疾病的严重程度。
Objective To investigate the clinical significance of PD-1 and B7-H1 gene expression in peripheral blood mononuclear cells from patients with oral lichen planus (OLP) and to investigate the clinical significance of PD-1 and its ligand B7- In blocking the B7-H1 / PD-1 signaling pathway on T cell proliferation and cytokine secretion to explore its relationship with the occurrence and development of OLP. Methods From January 2014 to January 2017, 300 OLP patients (OLP group) and 300 healthy adults (control group) were selected as the study subjects. Appraisal of disease status in patients with OLP using the Reticulo-Atrophy-Erosion (RAE) scoring system. Peripheral blood mononuclear cells were isolated by density gradient centrifugation. The expression of B7-H1 and PD-1 on T cells were detected by flow cytometry. Serum levels of IFN-γ, IL-2, IL-4, IL-10 and soluble PD-1 (s PD-1) were measured by enzyme linked immunosorbent assay (ELISA) After the mononuclear cells were removed by adherent method, the lymphocytes of OLP patients were mixed with healthy adult lymphocytes and anti-B7-H1 monoclonal antibody (anti-B7-H1m Ab) and anti-PD-1 monoclonal antibody -PD-1m Ab) blocked the B7-H1 / PD-1 signaling pathway and observed its effect on T cell proliferation and cytokine secretion. Results The expression of B7-H1 and PD-1 on peripheral blood T lymphocytes in OLP group was significantly higher than that in control group (all P <0.01). The expression level of B7-H1 in peripheral blood T lymphocyte of OLP group was positively correlated with RAE score (r = 0.510, P <0.01), while the expression of PD-1 was not correlated with RAE score system (r = 0.041, P> 0.05). The levels of Th1 cytokines IFN-γ and IL-2 in OLP group were significantly higher than those in control group (all P <0.01), while the levels of IL-4 and IL-10 in Th2 cytokines were significantly lower than those in control group (all P <0.01) . Serum levels of s PD-1 in OLP group and control group had no significant difference (P> 0.05). The antiproliferative effect of anti-B7-H1m Ab and anti-PD-1m Ab on B7-H1 / PD-1 signaling pathway was significantly higher than that of the pre-blocking OLP group (P <0.05) The secretion of IL-2 was significantly higher than that in the OLP group (P <0.05, P <0.01), indicating that B7-H1 / PD-1 signaling pathway can inhibit the proliferation of OLP T cells and the expression of IFN-γ and IL-2 The secretion. Conclusions The abnormal expression of B7-H1 / PD-1 in peripheral blood T lymphocytes in OLP patients may affect the occurrence and development of OLP by negatively regulating T cell proliferation and cytokine secretion in OLP patients. The expression level of B7-H1 on peripheral blood T lymphocytes in OLP patients is To monitor the severity of the disease.