乙型肝炎患者外周血Ⅱ型树突状细胞表型和功能鉴定及其意义的研究

目的　探讨Ⅱ型树突状细胞 (typeII dendriticcells,DC2 )数量和功能的变化及其与乙型肝炎 (乙肝 )患者病程、淋巴细胞亚群和HBVDNA病毒载量的关系。方法　采用流式细胞分析技术对 10 3例乙肝患者 (包括HBV携带者 11例 ,慢性乙肝轻度 35例 ,中度 33例 ,重度 13例 ,急性乙肝11例 )和 2 5例健康人 (对照组 )的外周血DC2进行检测 ,同时检测患者血常规 ,计算出DC2绝对数 ;用体外灭活的 1型单纯疱疹病毒 (HSV 1)刺激外周血单个核细胞 (PBMC)并与PBMC进行 2 4h共培养 ,检测上清中DC2的α干扰素产生能力 ;统计分析DC2数量、功能变化及其临床意义。结果　对照组外周血DC2占PBMC比例为 0 32 %± 0 13% ,绝对数为 (7 2± 2 4 ) 10 6个 /L。与对照组比较 ,HBV携带者DC2的比例 (0 32 %± 0 12 % )和绝对数 (6 9± 3 4 ) 10 6个 /L无明显下降 ;急性乙肝患者的DC2比例和绝对数下降 ,但无统计学意义 ;而慢性肝炎患者DC2的比例和绝对数随病程加重而降低 ,中、重度肝炎的DC2下降较轻度更为显著。DC2产生α干扰素的功能在急、慢性乙肝患者及HBV携带者均降低 ,各组患者之间差异不显著 ;DC2的数量与HBVDNA病毒载量未发现相关性 ,但与自然杀伤 (NK)细胞及CD8+ T数量高低存在正相关。结论　慢性乙肝患者外周血DC2数量和

Identification of phenotype and Interferon-alpha-producing capability of circulating type II dendritic cells and its clinical implication in HBV-infected patients

OBJECTIVE: To investigate the number, phenotype, and interferon-alpha (INF-alpha) of type II dendritic cells (DC2) in persons with hepatitis B and evaluate the role of DC2 subset in the immunopathogenesis of chronic HBV infection. METHODS: Peripheral blood was extracted from 103 hepatitis B (HB) virus-infected persons, including 11 cases of HB virus (HBV)-infected persons, 11 cases of acute HB, 81 cases of chronic HB, and 11 cases of asymptomatic HBV infection, and 25 healthy blood donors used as controls. Flow cytometry was used to calculate the number and the phenotype of circulating DC2. Ultraviolet-inactivated herpes simplex virus (HSV)-1 was added into the suspension of peripheral blood mononuclear cells (PBMCs) and then co-cultured for 24 hours to stimulate the production of INF-alpha by DC2 that was examined by ELISA assay. RESULTS: The number of DC2 in patients with chronic HB was 3.3 +/- 1.0 10(6)/L, significantly lower than that in the healthy controls (7.2 +/- 2.4 10(6)/L, P < 0.01). However, the number of DC2 was not significantly different between any other groups. The proportion of GS2 to PBMCs in the patients with chronic HB was 1.12 +/- 1.13 approximately 0.22 +/- 0.10, all significantly lower than that in the healthy controls (0.32% +/- 0.13%, P < 0.01). However, the proportion of GS2 to PBMCs was not significantly different between any other groups. The decrease of number of DC2 and that of proportion of DC2 to PBMCs in patients with chronic HB were related with the progress of disease. The INF-alpha concentration in the suspensions of PBMCs of different groups without stimulation by HSV-1 were low and there was no significant difference in INF-alpha concentration between different groups. The INF-alpha concentration in the suspension of PBMCs of healthy controls was 789 +/- 82 pg/ml, significantly higher than those of the patients with acute HB (161 +/- 36 pg/ml) and the patients with chronic HB (183 +/- 113 pg/ml, 147 +/- 39 pg/ml, and 156 +/- 39 pg/ml, all P < 0.05). However, there was no significant difference between the patient groups (all P > 0.05). CONCLUSION: The number and INF-alpha producing function of DC2, and the numbers of NK cells and CD8+ T cells in peripheral blood of patients with chronic HB decrease significantly, which results the deficiency of HBV-specific immune response.