亚硒酸钠作用的树突状细胞体外抗乙型肝炎病毒效应

Cytotoxic-T-lym phocytemediated anti-HBV effects of dendritic cells sensitized by sodium selenite in vitro

目的:探讨亚硒酸钠作用的树突状细胞(DCs)体外诱导自体淋巴细胞特异性抗乙型肝炎病毒的效应.方法:从乙型肝炎患者外周单个核细胞(PBMC)中诱导DCs,在DCs成熟前加入营养浓度的亚硒酸钠作用,将成熟后的DCs与自身淋巴细胞体外共培养,3d后收集淋巴细胞,分组(乙肝组,加硒组)加入2.2.15细胞培养液中,分别收集24,48和72h的培养上清,检测其表面抗原(HBsAg)和e抗原的分泌以及其与淋巴细胞共培养产生IFN-γ的含量.结果:经亚硒酸钠作用后的DCs组和正常对照组的DCs与自身淋巴细胞共培养产生IFN-γ的浓度,以及在24,48和72h对2.2.15细胞培养上清中HBeAg分泌的抑制效应明显高于乙肝组(34.22±3.17ng/L,3839±2.43ng/L vs 21.47±2.24ng/L;25.90%±1.85%,23.03%±1.51% vs 18.05%±3.64%;37.26%±5.11%,36.88%±5.92% vs 29.52%±2.63%;38.76%±4.00%,40.76%±5.04% vs 35.59%±3.09%,P<0.05).结论:体外经亚硒酸钠作用后的乙肝患者的DC功能可在一定程度上恢复,增强自身淋巴细胞毒性作用并可提高产生IFN-γ的含量,可有效的起到抗乙肝病毒的效应,为以后基于DC的慢性乙型肝炎的免疫治疗提供新思路.

AIM： To investigate the cytotoxic-T-lymphocyte （CTL）-mediated anti-HBV effects of dendritic cells （DCs） from patients with HBV sensitized by sodium selenite in vitro. METHODS： DCs isolated from peripheral blood mononuclear cells of HBV patients were cultured and proliferated in vitro. They were divided into the sodium-selenite-stimulated group and the HBV group. DCs were stimulated with pure sodium selenite before maturation and then co-cultured with autolymphocytes after DC maturation. Lymphocytes were harvested 3 d later and added to the supernatant of 2.2.15 cell cultures. Supernatants were collected at 24, 48 and 72 h. ELISA was used to detect hepatitis B e antigen （HBeAg） and hepatitis B surface antigen （HBsAg）, and to measure the concentration of interferon （IFN）-γ in the culture Supernatants. RESULTS： Proliferation, cytotoxicity and IFN-γ concentration of autolymphocytes were markedly enhanced in the sodium-selenite-stimulated DCs and control groups compared with the HBV group （34.22 ± 3.17 ng/L and 38.39 ± 2.43 ng/L vs 21.47 ± 2.24 ng/L, P 〈 0.05）. Compared with the liver group, CTLs activated in the sodium selenite and control groups had specific inhibitory effects on the expression of HBeAg in 2.2.15 cell supernatants （25.90% ± 1.85% and 23.03% ± 1.51% vs 18.05% ± 3.64%; 37.26% ± 5.11% and 36.88% ±5.92% vs 29.52% ± 2.63%; 38.76%± 4.00% and 40.76% ± 5.04% vs 35.59% ± 3.09%; P 〈 0.05）. CONCLUSION： Human DCs sensitized by sodium selenite in vitro appear to efficiently enhance cytotoxicity and increase the level of IFN-γ, and effectively inhibit the expression of HBeAg in 2.2.15 cell supernatant. This may indicate a new route for therapy against chronic hepatitis B.