不同病毒载量的慢性乙型肝炎患者外周血树突状细胞的表型和功能

目的观察研究不同HBV DNA载量的CHB患者外周血DC表型及功能变化,探讨HBV在DC成熟障碍中的作用机制。方法筛选血清HBV DNA>10~5拷贝/ml且无其他肝脏疾病及自身免疫性疾病的CHB患者28例。所有病例予核苷类似物抗病毒治疗24周。治疗前后测定外周血HBV DNA并经肝穿刺明确肝组织病理状态。将治疗前患者设为高载量组(HBV DNA>10~5拷贝/ml),共28例;治疗后患者设为低载量组(HBV DNA<10~3拷贝/ml),共25例。从患者外周血分离单核细胞,体外诱导培养DC。用流式细胞仪测定DC表型,MTT法测定DC对同种异体淋巴细胞的刺激增殖作用,ELISA法测定DC分泌IL-12和IL-10的量。同时以10名健康人作对照。结果DC在体外经细胞因子的刺激可明显增殖,但CHB患者DC的扩增数量、速度低于正常人。DC表面标记:正常人的HLA-DR、CD86、CD80和CD83表达阳性率均大于80%,显著高于两组CHB患者;不同HBV DNA载量的两组CHB患者间差异无统计学意义。两组CHB患者的DE在混合淋巴细胞反应中的刺激能力差异无统计学意义,但明显低于正常对照组。CHB患者和正常人DC上清液中IL-10的量,各组间差异无统计学意义;高载量组与低载量组纯DC培养和混合淋巴细胞反应上清液中IL-12量的差异无统计学意义,而分别明显低于正常人DC。结论在HBV持续感染期间,CHB患者DC的表型变化及功能下调与外周血HBV DNA载量间差异无统计学意义。

Phenotypes and functions of dendritic cells derived from peripheral blood monocytes of chronic hepatitis B patients with different HBV DNA loads

Objective To investigate the phenotypes and functions of peripheral blood monocyte derived dendritic cells(DC)of chronic hepatitis B(CHB)patients with different HBV DNA loads.Methods Twenty-eight CHB patients were included in this study.All patients were treated with nucleoside analogues (lamivudine or LdT or adeforvir)for 24 weeks.Peripheral blood HBV DNA loads and liver biopsies were assessed before and after the treatment.The patients were divided into two groups according to their periph- eral blood HBV DNA loads:a high-load group with HBV DNA loads higher than 10~5 copies/ml,and a low- load group with HBV DNA loads lower than 10~3 copies/ml.Ten healthy people were included as controls. Peripheral blood DC of each subject was enriched.The phenotypes of DC were subjected to flow cytometric analysis.The lymphocyte allo-stimulatory capacity of DC was evaluated through MTT assay.IL-10 and IL- 12 production were quantified by ELISA.Results DC proliferated successfully when stimulated by cytokines in vitro;however,DC of the CHB patients proliferated much slower than those of the healthy controls.The expression of DC surface molecules such as HLA-DR,CD86,CD80 and CD83 had a positive rate of over 80% in the normal population.However in our CHB patients they showed lower than normal expressions, especially the HLA-DR,CD86,CD80 and CD83,but the differences were not significant between the two groups with different virus loads.The stimulatory capacity of the DC in mixed lymphocyte reac- tion showed no difference between the two groups of patients,but both were lower than that of the healthy controls.The production of IL-12 and IL-10 also decreased significantly in the patients.Con- clusions Peripheral DC of CHB patients have some defects in their phenotypes and their stimulatory capacity.The changes in phenotypes and down-regulation of the functions are not relevant to periph- eral HBV DNA loads of the patients.