本文選題：乙型肝炎病毒 + 樹突狀細胞��； 參考：《復(fù)旦大學(xué)》2010年碩士論文

【摘要】： 目的： 通過觀察乙型肝炎病毒干預(yù)后小鼠肝臟樹突狀細胞干擾素調(diào)節(jié)因子3表達的變化,探討HBV持續(xù)感染分子免疫機制。 方法： 采用細胞濾網(wǎng)過濾、Percoll密度梯度離心和免疫磁珠分選法分離肝臟樹突狀細胞,并用粒細胞—巨噬細胞集落刺激因子和白介素4誘導(dǎo)培養(yǎng)。將樹突狀細胞分為兩組：一組與HBV共培養(yǎng),另一組加入等量的細胞培養(yǎng)液作為正常對照組。24h后,兩組均加入Poly I:C刺激,收集不同時間點細胞和上清,留細胞培養(yǎng)液作為空白對照,用western blot檢測IRF3表達,用ELISA法檢測上清中IFN-β濃度。 結(jié)果： 1.DC與HBV共培養(yǎng)前,IRF3表達弱。用poly I:C刺激DC后,IRF3表達明顯增加,呈時相性,在2-6h達到高峰。DC與HBV共培養(yǎng)后,再用poly I:C刺激,IRF3表達的時相性不明顯,未見明顯升高。DC表達IRF3的能力與病毒載量有關(guān),高病毒載量能明顯抑制IRF3表達,病毒載量低于106 copies／ml時差異不明顯。 2.0h時HBV組培養(yǎng)液上清IFN-β濃度(12.38±3.71pg／ml)與正常對照組(10.83±4.11 pg／ml)相比,差異無統(tǒng)計學(xué)意義(t=0.8398,P0.05)。Poly工：C刺激后6h,HBV組培養(yǎng)液上清IFN-β濃度(88.67±9.01 pg／ml)與正常對照組(137.68±12.28 pg／ml)相比,差異有統(tǒng)計學(xué)意義(t=9.653,P0.01)。Poly I:C刺激后24h,HBV組培養(yǎng)液上清IFN-β濃度(69.89±5.80 pg／ml)與正常對照組(72.25±8.61 pg／ml)相比,差異無統(tǒng)計學(xué)意義(t=0.6820,P0.05)。 結(jié)論： 1.用poly I:C刺激正常DC后,IRF3表達增加,呈時相性。 2.HBV干預(yù)后,用poly I:C刺激DC后,IRF3表達增加不明顯。高載量HBV能明顯抑制肝臟樹突狀細胞IRF3的表達。 3.HBV干預(yù)后,肝臟樹突狀細胞IRF3下游IFN-β表達降低。
[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 鹵3.71pg/ml) compared with that in the control group (10.83 鹵4.11 pg/ml). The concentration of IFN- 尾 in supernatant of HBV-treated group (88.67 鹵9.01 pg/ml) was significantly higher than that of control group (137.68 鹵12.28 pg/ml). The concentration of IFN- 尾 in the supernatant of HBV-treated group (69.89 鹵5.80 pg/ml) was not significantly different from that in the control group (72.25 鹵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.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

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相關(guān)期刊論文 前2條

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