慢性乙型肝炎患者外周血树突状细胞表型及抗原提呈能力与HBV载量的关系  

Relationships between the phenotypes, the capacity of antigen presenting of peripheral blood dendritic cells and the loads of HBV in patients with chronic hepatitis B

作　　者：钱峰[1] 贺永文[2] 朱传武[1] 王海燕[1] 吴妹英[1] 李明[1] 

机构地区：[1]苏州市第五人民医院传染科,215007 [2]华中科技大学同济医学院附属协和医院

出　　处：《肝脏》2005年第1期8-10,共3页Chinese Hepatology

摘　　要：目的　探讨慢性乙型肝炎 (CHB)患者外周血树突状细胞 (DCs)表型及抗原提呈能力与HBV载量的关系。方法　采集 2 3例CHB患者和 8例健康人的抗凝外周静脉血 ,分离外周血单个核细胞 (PBMCs) ,在重组人白细胞介素 4和重组人粒细胞 巨噬细胞集落刺激因子的作用下培养使DCs增殖、成熟 ,以间接免疫荧光流式细胞技术分别检测DCs表面CD80、CD86、HLA DR及ICAM 1的表达 ;将培养成熟的DCs与HBsAg共同孵育 ,用丝裂霉素C处理后再与自体PBMCs共同培养 ,在培养结束前 12h加入3H TDR ,收集细胞 ,以 β液闪计数仪测定cpm值 ;同期用定量聚合酶链反应技术测定CHB患者外周血HBV载量。结果　患者DCs表面CD86、HLA DR和ICAM 1的表达水平及DCs的抗原提呈能力均显著低于健康对照组 ;CD80、CD86、HLA DR及ICAM 1的表达与HBV载量呈显著负相关 (分别为P <0 .0 1,P <0 .0 1,P <0 .0 0 1和P <0 .0 0 1) ;DCs的抗原提呈能力也与HBV载量呈显著负相关 (为P <0 .0 0 1)。结论　CHB患者外周血DCs的成熟和功能存在障碍 ,DCs的抗原提呈能力与血液中HBV的载量密切相关 。Objective To Study the relationships between the phenotypes, the capacity of antigen presenting of peripheral blood dendritic cells(DCS) and the loads of HBV in patients with chronic hepatitis B. Methods The peripheral blood samples were obtained from 23 patients with chronic hepatitis B (CHB) and 8 healthy individuals. The peripheral blood mononuclear cells (PBMCs) were isolated. DCs were isolated from peripheral blood mononuclear cells and enriched by culturing with recombinant human interleukin-4 (rhIL-4) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF), then DCs were harvested and the molecules of CD80, CD86, HLA-DR as well as ICAM-1 on DCs were tested by indirect immunofluorescence flow cytometry. The matured DCs after culture were incubated with HBsAg,then after treated with mytomycin C, DCs were mixed with autogenous PBMCs and cocultured. () 3H-TDR was added to the culture 12 hours before stopping the culture, and all the cells were collected and cpm values were measured. The loads of HBV were measured with fluorescence quantitative polymerase chain reaction (FQ-PCR).Results The levels of CD86, HLA-DR and ICAM-1 expressed on DCs, and the capacity of antigen presenting of DCs in patients' group were remarkably lower than those in control group, respectively. A significantly negative correlation was noted between the expressed level of CD80, CD86, HLA-DR, ICAM-1 on DCs and the loads of HBV (P<0.01, P<0.01, P<0.001 and P<0.001, respectively), and also could be found between antigen presenting capacity of DCs and the HBV loads. (P<0.001). Conclusion The function and maturation of DCs were injured in patients with CHB. The functional status of DCs may closely correlate with the loads of HBV, and thus may exert important influence on the clearance of HBV.

关 键 词：抗原提呈能力 慢性乙型肝炎患者 HBV 树突状细胞表型 重组人粒细胞-巨噬细胞集落刺激因子 载量 重组人白细胞介素4 ICAM-1 HLA-DR 外周血树突状细胞 外周血单个核细胞 聚合酶链反应技术 ^3H-TDR 流式细胞技术 间接免疫荧光 

分 类 号：R512.6[医药卫生—内科学]