乙型肝炎病毒干预后小鼠肝脏树突状细胞干扰素调节因子3表达变化的研究

发布时间：2018-06-28 17:37

本文选题：乙型肝炎病毒 + 树突状细胞　；参考：《复旦大学》2010年硕士论文

【摘要】： 目的：通过观察乙型肝炎病毒干预后小鼠肝脏树突状细胞干扰素调节因子3表达的变化,探讨HBV持续感染分子免疫机制。方法：采用细胞滤网过滤、Percoll密度梯度离心和免疫磁珠分选法分离肝脏树突状细胞,并用粒细胞—巨噬细胞集落刺激因子和白介素4诱导培养。将树突状细胞分为两组：一组与HBV共培养,另一组加入等量的细胞培养液作为正常对照组。24h后,两组均加入Poly I:C刺激,收集不同时间点细胞和上清,留细胞培养液作为空白对照,用western blot检测IRF3表达,用ELISA法检测上清中IFN-β浓度。结果： 1.DC与HBV共培养前,IRF3表达弱。用poly I:C刺激DC后,IRF3表达明显增加,呈时相性,在2-6h达到高峰。DC与HBV共培养后,再用poly I:C刺激,IRF3表达的时相性不明显,未见明显升高。DC表达IRF3的能力与病毒载量有关,高病毒载量能明显抑制IRF3表达,病毒载量低于106 copies／ml时差异不明显。 2.0h时HBV组培养液上清IFN-β浓度(12.38±3.71pg／ml)与正常对照组(10.83±4.11 pg／ml)相比,差异无统计学意义(t=0.8398,P0.05)。Poly工：C刺激后6h,HBV组培养液上清IFN-β浓度(88.67±9.01 pg／ml)与正常对照组(137.68±12.28 pg／ml)相比,差异有统计学意义(t=9.653,P0.01)。Poly I:C刺激后24h,HBV组培养液上清IFN-β浓度(69.89±5.80 pg／ml)与正常对照组(72.25±8.61 pg／ml)相比,差异无统计学意义(t=0.6820,P0.05)。结论： 1.用poly I:C刺激正常DC后,IRF3表达增加,呈时相性。 2.HBV干预后,用poly I:C刺激DC后,IRF3表达增加不明显。高载量HBV能明显抑制肝脏树突状细胞IRF3的表达。 3.HBV干预后,肝脏树突状细胞IRF3下游IFN-β表达降低。
[Abstract]:Aim: to investigate the molecular immune mechanism of hepatitis B virus (HBV) persistent infection by observing the expression of interferon regulatory factor 3 (IFN 3) in mouse liver dendritic cells. Methods: hepatic dendritic cells were isolated by Percoll density gradient centrifugation and immunomagnetic bead sorting, and cultured with granulocyte-macrophage colony stimulating factor and interleukin-4. Dendritic cells were divided into two groups: one group was co-cultured with HBV, the other group was treated with the same amount of cell culture medium as the normal control group for 24 h, both groups were stimulated with Poly I: C to collect cells and supernatants at different time points. The expression of IRF3 was detected by western blot and the concentration of IFN- 尾 in supernatant was detected by Elisa. Results: 1. The expression of IRF3 in DC and HBV coculture was weak. The expression of IRF3 in DC stimulated by poly I: C was obviously increased, and the expression of IRF3 was not obvious after the peak of poly I: C was reached between 2 and 6 hours, and the ability of expressing IRF3 in DC was not related to the viral load. The expression of IRF3 was significantly inhibited by high viral load, but there was no significant difference when the viral load was lower than 106 copies/ml. The concentration of IFN- 尾 in the supernatant of HBV-treated medium at 2.0 h was (12.38 卤3.71pg/ml) compared with that in the control group (10.83 卤4.11 pg/ml). The concentration of IFN- 尾 in supernatant of HBV-treated group (88.67 卤9.01 pg/ml) was significantly higher than that of control group (137.68 卤12.28 pg/ml). The concentration of IFN- 尾 in the supernatant of HBV-treated group (69.89 卤5.80 pg/ml) was not significantly different from that in the control group (72.25 卤8.61 pg/ml) (t = 0.6820, P 0.05). Conclusion: 1. The expression of IRF3 in normal DC was increased after stimulation with poly I: C, and the expression of IRF3 was not significantly increased after poly I: C stimulated DC. The expression of IRF3 in hepatic dendritic cells was significantly inhibited by high dose of HBV. 3. The expression of IFN- 尾 in liver dendritic cells downstream of IRF3 was decreased after HBV intervention.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

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