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目的探讨HBcAg和HBeAg对小鼠骨髓源性树突状细胞功能的影响。方法分离C57BL/6小鼠骨髓细胞,用rm GM-CSF和rm IL-4体外诱导为树突状细胞(DCs)后,按照干预条件分为对照组、HBcAg组和HBeAg组。混合淋巴细胞反应(MLR)检测DCs刺激T淋巴细胞增殖能力,酶联免疫法(ELISA)检测培养上清中IL-12、IL-10和IDO的分泌水平,Western blot法检测Akt磷酸化水平,设置LY294002组为阳性对照探讨细胞因子分泌的可能调节机制。结果 HBc Ag组上清液中IL-12水平以及DCs刺激淋巴细胞增殖能力较对照组明显升高(P<0.01)。HBe Ag组上清液中IL-12水平以及DCs刺激淋巴细胞增殖能力较对照组明显降低(P<0.05),而IL-10和IDO的水平较对照组明显升高(P<0.01)。HBeAg组Akt的磷酸化水平较对照组明显升高(P<0.05)。LY294002组Akt的磷酸化水平较HBeAg组明显降低(P<0.05),并且上清液中IL-12水平较HBeAg组明显升高(P<0.01),IL-10和IDO的水平较HBeAg组明显降低(P<0.01)。结论相对于HBc Ag的正性作用,HBe Ag通过PI_3K-Akt信号通路调节DCs细胞因子分泌,减弱DCs的免疫功能,这可能是乙型肝炎慢性化的机制之一。
Objective To investigate the effect of HBcAg and HBeAg on the function of bone marrow-derived dendritic cells in mice. Methods The bone marrow cells of C57BL / 6 mice were isolated and induced with rm GM-CSF and rm IL-4 as dendritic cells (DCs) in vitro. The cells were divided into control group, HBcAg group and HBeAg group according to the intervention conditions. IL-12, IL-10 and IDO secretion in culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA), Akt phosphorylation was detected by Western blot, and the proliferation of T lymphocytes was detected by mixed lymphocyte reaction (MLR) LY294002 group was set as a positive control to explore the possible regulatory mechanism of cytokine secretion. Results The level of IL-12 in supernatant of HBc Ag group and the ability of DCs to stimulate lymphocyte proliferation were significantly higher than those in control group (P <0.01). The level of IL-12 in supernatant of HBe Ag group and the ability of DCs to stimulate lymphocyte proliferation were significantly lower than those in control group (P <0.05), while the levels of IL-10 and IDO were significantly higher than those in control group (P <0.01). Akt phosphorylation in HBeAg group was significantly higher than that in control group (P <0.05). The phosphorylation level of Akt in LY294002 group was significantly lower than that in HBeAg group (P <0.05), and the level of IL-12 in supernatant was significantly higher than that in HBeAg group (P <0.01). The levels of IL-10 and IDO in LY294002 group were significantly higher than those in HBeAg group Decreased (P <0.01). Conclusions HBe Ag regulates the cytokine secretion of DCs through the PI_3K-Akt signaling pathway and attenuates the immunological function of DCs, which may be one of the mechanisms of chronic hepatitis B infection.