中文摘要：

目的探讨HBeAg对慢性乙型肝炎（CHB）患者外周血单个核细胞（PBMC）功能的调节作用。方法以重组的HBeAg体外刺激CHB患者和健康志愿者的PBMC，用流式细胞术和酶联免疫吸附试验法检测其刺激前后Th1／Th2型细胞因子的变化情况，并观察HBeAg对CHB患者PBMC表面细胞程序性死亡受体（PD）1及其配体（PD—L）l表达的影响。两组间资料比较采用独立样本t检验IPD—I／PD—L1表达水平与HBVDNA拷贝数的相关性采用Spearman相关分析。结果HBeAg刺激后可使HBeAg阴性CHB患者和健康志愿者CD3＋CD4＋T淋巴细胞内干扰素（IFN）Y表达水平（0．17％±0．08％与0．17％±0．04％）明显低于未刺激组（0．30％±0．16％与0．32％±0．12％），t值分别为-2．382和-4．190，P值均〈0．01;培养上清液中白细胞介素（IL）-6、IL—10和肿瘤坏死因子α含量明显高于未刺激组（HBeAg阴性CHB患者的t值分别为2．504，3．583和4．324，健康志愿者t值分别为3．542，6．246和5．273，P值均〈0．01）。HBeAg刺激PBMC后，HBeAg阴性CHB患者和健康志愿者CD14＋细胞表面PD—L1表达水平分别为13．02％±4．98％和3．10％±2．47％，明显高于未刺激组的5．89％±1．56％和0．97％±0．83％，t值分别为4．815和3．454，P值均〈0．05。基础状态下在HBeAg阳性CHB患者外周血中，CD3＋CD4＋T淋巴细胞内IFNY表达水平为0．23％±0．09％，明显低于HBeAg阴性CHB患者和健康志愿者的0．34％±0．15％和0．35％±0．09％（t=-3．177，P〈0．01；t=-4．541，P〈0．01）；而IL-4表达水平为0．39％±0．16％，明显高于HBeAg阴性CHB患者和健康志愿者的0．26％±0．12%和0．23％±0．12％，t值分别为3．382和4．393，P值均〈0．01。基础状态下在HBeAg阳性CHB患者外周血中，CD3＋T淋巴细胞表面PD-1和PD—L1表达水平明显高于HBeAg阴性CHB患者及健康志愿者（P值均〈0．01），CD14＋T淋巴细胞?

英文摘要：

Objective To study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).Methods PBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg-and HBeAg+) and healthy controls,and cultured with recombinant HBeAg.The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants.Comparisons between two groups were made by the independent-samples t-test;the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spcarman's correlation analysis.Results Exposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNα for both the CHB patients' and healthy controls' samples (t =2.382 and-4.190 respectively,P ＜ 0.01).For the HBeAg-CHB patients' and healthy controls' samples,the HBeAg exposure led to increased levels of secreted cytokines IL-6,IL-10 and TNFα (t =2.504,3.583 and 4.324,P ＜ 0.01 and t =3.542,6.246 and 5.273,P ＜ 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t =4.815 and 3.454,P ＜ 0.05 respectively).Compared to the HBeAg-negative CHB patients' and healthy controls' samples,the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNα (t =-3.177 and-4.541,P ＜ 0.01 respectively),but significantly higher levels of secreted IL-4 (t =3.382 and 4.393,P ＜ 0.01 respectively),ofCD3+ T cells-specific expression of PD-1/PD-L1 (t =4.755,2.942 and 4.518,4.595,P ＜ 0.01 respectively),and of CD14+ T cells-specific expression of PD-L1 (t =5.092 and 5.473,P ＜ 0.01 respectively).The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg-CHB patients than from the healthy controls (t =3.214,P ＜ 0.01).Conclusion HBeAg was able to down-regulate the production ofTh1-type eytokines (IFNγ),and up-regulate the secretion of Th2-type cytokines (IL-6,IL-10) and the