人树突状细胞体外经HBsAg刺激后的抗病毒作用

目的观察人树突状细胞(DCs)经表面抗原(HBsAg)刺激、体外诱导自身特异性T淋巴细胞增殖后,对2.2.15细胞中HBeAg和HBsAg的特异性免疫抑制作用。方法 用粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白介素-4(IL-4)和肿瘤坏死因子(TNF-a)分化、诱导人外周血PBMC中的DCs,在DCs成熟前加入纯的HBsAg刺激,将成熟后的DCs体外与自身T淋巴细胞共培养,同时不加HBsAg刺激的DCs与T细胞共培养、T细胞加纯的HBsAg共培养以及单纯T细胞作对照,5 d后收集T细胞,分组加入2.2.15细胞培养液中,分别收集第1天、3天、5天和7天的培养上清液,检测其HBeAg和HBsAg的分泌情况。结果经抗原刺激后的DCs可以有效提呈病毒抗原,正常人与慢性乙肝患者负载抗原后的DCs刺激T淋巴细胞增殖的能力cpm分别为(46 700±7 850)和(38 628±5 427)]明显高于未负载抗原的DCscpm分别为(40 450±4 645和33 924±4 498)]及对照组PBMCcpm分别为(5 947±476)和(5 089±233)],P<0.01。负载抗原的DCs有强烈的免疫应答活性,并且其免疫刺激能力似乎与负载的抗原量成正比;经抗原刺激激活的T细胞可以有效地抑制HBeAg的表达,但对HBsAg未发现有明显的抑制作用。结论体外经HBsAg刺激后的DCs可有效地提呈病毒抗原,并可进一步激活T细胞产生,同时能显著地抑制2.2.15细胞上清

Antiviral effect of human CTLs activated by HBsAg-stimulated dendritic cells in vitro

Objective To investigate the ability of human dendritic cells ( DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant.Methods DCs were prepared from peripheral blood mononuclear cells induced with granulocyte-macrophage colony-stimulating factor( GM-CSF) and interleukin-4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2. 15 cell with stimulated T lymphocytes was collected on day 1, day 3,day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method. Results DCs after antigen stimulation had a strong ability to present antigen and induce immune activation , DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P <0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant, but not on the expression of HBsAg. Conclusion Human dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLa to HBV, which can efficiently inhibite the expression of HBeAg in 2.2.15 cell supernatant. DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future.