慢性乙型肝炎患者树突状细胞分离培养及其功能测定

目的探讨树突状细胞（DCs）在慢性乙型肝炎发病过程中的功能特点，为进一步研究慢性乙型肝炎的发病机制提供科学依据。方法对10例慢性乙型肝炎患者、10例乙型肝炎病毒（HBV）携带者及10例正常对照者的外周血DCs进行体外培养，应用流式细胞技术检测DCs表面分子HLA-DR、CD80、CD86的表达，并采用酶联免疫吸附试验（ELISA）检测DCs上清液白细胞介素（IL）-12的水平，比较DCs在慢性乙型肝炎不同感染阶段的功能特点。结果培养7天的DCs在透射电镜下观察，DCs充满每个视野，细胞体积较大，细胞表面突起较丰富，胞浆内粗面内质网及溶酶体丰富，线粒体结构较清楚，核大而圆，核膜清晰，染色质分布较均匀。正常对照组的HLA-DR、CD80和CD86的表达率均〉54％，而慢性乙型肝炎及HBV携带者组上述DCs表面分子的表达率普遍降低，与正常对照组比较，差异有显著性（P〈0．01），慢性乙型肝炎与HBV携带者组之间的差异无统计学意义（P〉0．05）。培养第7天DCs上清IL-12的表达水平在慢性乙型肝炎、HBV携带者组明显低于正常对照组（P〈0．05），而慢性乙型肝炎与HBV携带者组之间无明显差异（P〉0．05）。结论慢性乙型肝炎患者及HBV携带者均存在DCs表型和功能的缺失。

Isolation and incubation of dendritic cell and determination of the function of dendritic cell from patients with chronic hepatitis B

Objective To evaluate the function and characteristics of dendritic cells(DCs) in the progress of chronic hepatitis B,so as to provide a scientific basis for further study on the pathogenesis of DCs in chronic hepatitis B.Methods DCs in peripheral blood mononuclear cells(PBMCs) of 10 chronic hepatitis B patients,10 HBV carriers,and 10 healthy controls were incubated in vitro,and the expression of HLA-DR,CD80 and CD86 were detected by flow cytometry,the level of IL-12 in supernatants was measured by ELISA.The functions and characteristics of DCs in different stages of chronic hepatitis B were compared.Results On the seventh day of incubation,the cells were observed by transmission electron microscope,volume of the cell was large with affluent process on surface,abundant rough surfaced endoplasmic reticulum and lysosome in cytoplasma,distinct mitochondria structure,big and round nucleus and clear nuclear membrane,evenly distributed chromatin.The results showed that the expression levels of HLA-DR,CD80 and CD86 in healthy control group were all>54%,and in patients and HBV carriers were decreased,there was significant difference compared with healthy control group(P<0.01),but there was no statistical significance between patients and HBV carriers(P>0.05).The levels of IL-12 in the supernatants in patients and HBV carriers were much lower than that in healthy controls(P<0.05),but there was no statistical significance between patients and HBV carriers(P>0.05).Conclusion DCs in chronic hepatitis B patients and HBV carriers have not only an immature phenotype,but also an impaired function.