發(fā)布時間：2018-03-02 05:17

  本文關(guān)鍵詞： Exosome 樹突狀細(xì)胞 HBV 免疫治療　出處：《福建醫(yī)科大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的 初步探索Exosome體外抗HBV特異性免疫活性。 方法 以細(xì)胞因子GM-CSF、IL-4及TNF-α培養(yǎng)健康人PBMC來源的DCs,通過流式細(xì)胞儀分析細(xì)胞表型判斷所收獲的DCs成熟度;采用多步驟離心法分離純化DCs培養(yǎng)上清中的Exosome,western blotting分析蛋白性質(zhì);以HBcAg刺激DCs,MTS法檢測收獲的HBcAg特異性DEXs刺激PBMC增殖的能力、ELISPOT法檢測HBcAg特異性DEXs刺激的PBMC產(chǎn)IFN-γ的能力。 結(jié)果 成熟DC表面HLA-DR、CD80/CD86及CD83分子的表達(dá)水平明顯高于未成熟DC者(分別為45.3±8.3% vs 16.5±6.8%、90.2±6.5% / 92.38±13.6% vs 6.3±4.5%/49.8±9.2%、83.53±10.8% vs 4.3±2.8%,P0.05);成熟DCs分泌的Exosome表達(dá)HLA-DR、CD86分子也強(qiáng)于未成熟DCs分泌者;淋巴細(xì)胞增殖實驗結(jié)果顯示HBcAg特異性DEXs體外刺激PBMC增殖能力強(qiáng)于未成熟者(吸光度0.904±0.003 vs 0.545±0.010,P0.001),但不如HBcAg特異性mDCs(0.904±0.003 vs 1.034±0.004,P0.05);HBcAg特異性DEXs刺激的PBMC產(chǎn)IFN-γ能力強(qiáng)于未成熟者(斑點數(shù)32±13 vs 14±2,P0.05),但也不如HBcAg特異性mDCs(32±13 vs 98±17,P≤0.01 )。 結(jié)論 HBcAg特異性mDEXs可有效激活自體PBMC增殖及分泌IFN-γ,其體外抗HBV特異性免疫活性強(qiáng)于未成熟者。
[Abstract]:Purpose. Objective: to explore the specific immunological activity of Exosome against HBV in vitro. Method. DCsderived from healthy human PBMC were cultured with cytokines GM-CSFF-IL-4 and TNF- 偽. The DCs maturity was determined by flow cytometry, and the protein properties of DCs supernatant were purified by multistep centrifugation. The ability of HBcAg specific DEXs to stimulate the proliferation of PBMC was detected by HBcAg stimulation of DCS MTS and the ability of PBMC stimulated by HBcAg specific DEXs to produce IFN- 緯 was detected by Elispot method. Results. The expression levels of HLA-DRN CD80 / CD86 and CD83 on mature DC were significantly higher than those of immature DC (45.3 鹵8.3% vs 16.5 鹵6.8 vs 90.2 鹵6.5% / 92.38 鹵13.6% vs 6.3 鹵4.5 鹵4.5T = 83.53 鹵10.8% vs 4.3 鹵2.8 P0.055.The Exosome expression of mature DCs was also stronger than that of immature DCs secreting. The results of lymphocyte proliferation test showed that HBcAg specific DEXs could stimulate the proliferation of PBMC in vitro better than the immature ones (absorbance: 0.904 鹵0.003 vs 0.545 鹵0.010) P 0.001, but it was not as good as HBcAg specific mDCs(0.904 鹵0.003 vs 1.034 鹵0.004 P0.05G DEXs to stimulate PBMC to produce IFN- 緯. The number was 32 鹵13 vs 14 鹵2, P 0.05, but less than HBcAg specific mDCs(32 鹵13 vs 98 鹵17 (P 鈮，

本文編號：1555171