摘要
目的探讨用HBsAg致敏的树突状细胞(DCs)体外诱导自体T淋巴细胞特异性抗乙型肝炎病毒的免疫效应。方法从乙型肝炎患者外周血单个核细胞(PBMC)中诱导DCs,在DCs成熟前加入纯的HBsAg刺激,将成熟后的DCs与自身T淋巴细胞体外共培养,5天后收集T细胞,分组加入2.2.15细胞培养液中,分别收集第1、3、5和7天的培养上清液,检测其HBeAg和HBsAg的分泌情况以及其与T淋巴细胞共培养产生的IFN-γ和IL-12的含量。结果经抗原刺激后的DCs刺激自身T淋巴细胞增殖可明显加强其细胞毒作用,同时其产生的IFN-γ和IL-12的浓度明显高于未经抗原刺激的DCs组(P<0.05)和对照组PBMC(P<0.01)。经抗原激活的T细胞可有效地抑制HBeAg的表达,但未发现对HBsAg有明显的抑制作用。结论体外经HBsAg刺激后的DCs激活的自身T细胞可明显地增强细胞毒作用并可提高产生IFN-γ和IL-12的含量,从而显著地抑制2.2.15细胞上清中HBeAg的表达,致敏后的DCs疫苗体外可有效地起到抗乙型肝炎病毒的效应。
Objective To investigate T cell-mediated anti-HBV effects of dendritic cells （DCs） sensitized by HBsAg in patients with HBV infection in vitro. Methods DCs isolated from peripheral blood mononuclear cells of HBV patients were cultured and proliferated in vitro, and then were stimulated with pure HBsAg before DCs maturation, and co-cultured with auto-T lymphocytes after DCs maturation. The T lymphocytes were then harvested 5 days later and added into the supematant of 2.2. 15 cell cultures. The supernatants were collected on 1st, 3rd, 5th and 7th day, respectively. ELISA was employed to detect FIBeAg and the secretion of HBsAg, as well as to measure the concentration of IFN-γ and IL-12 in culture supematents, Results The proliferation and cytotoxicity of auto-T cells were markedly en- hanced in the antigen stimulated DCs group compared with that of DCs without loaded antigen and only T lymphocyte group （P〈0. 01）. Concentration of IFN-γ and IL-12 increased in HBsAg pulsed DCs group compared with that of unpulsed DCs group （ P〈0. 05） and control group （P〈0. 01）. The ability of DCs to be stimulated bad a positive relation to the dosage of loaded antigenen; CTLs activated by the antigen bad a specific inhibitory effect on the expression of HBeAg in 2. 2. 15 cell supematant, but not on the expression of HBsAg. Conclusion Human DCs sensitized by HBsAg in vitro may efficiently enhance the cytotoxicity and increase the contents of IFN-γ and IL-12, effectively inhibits the expression of HBeAg in 2.2. 15 cell supematant. Sensitized DC,s vaccine may probably be used in antiviral immunological therapy for chronic hepatitis B virus infected patients in future.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2007年第1期39-42,共4页
Medical Journal of Chinese People's Liberation Army
关键词
树突细胞
抗病毒作用
免疫耐受
肝炎
乙型
细胞因子类
denritic cells
antiviral function
immune tolerance
chronic hepatitis B
cytokines
