Exosome用于抗HBV免疫治疗的实验研究

发布时间：2018-03-02 05:17

  本文关键词： Exosome 树突状细胞 HBV 免疫治疗　出处：《福建医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 目的 初步探索Exosome体外抗HBV特异性免疫活性。 方法 以细胞因子GM-CSF、IL-4及TNF-α培养健康人PBMC来源的DCs,通过流式细胞仪分析细胞表型判断所收获的DCs成熟度;采用多步骤离心法分离纯化DCs培养上清中的Exosome,western blotting分析蛋白性质;以HBcAg刺激DCs,MTS法检测收获的HBcAg特异性DEXs刺激PBMC增殖的能力、ELISPOT法检测HBcAg特异性DEXs刺激的PBMC产IFN-γ的能力。 结果 成熟DC表面HLA-DR、CD80/CD86及CD83分子的表达水平明显高于未成熟DC者(分别为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表达HLA-DR、CD86分子也强于未成熟DCs分泌者;淋巴细胞增殖实验结果显示HBcAg特异性DEXs体外刺激PBMC增殖能力强于未成熟者(吸光度0.904±0.003 vs 0.545±0.010,P0.001),但不如HBcAg特异性mDCs(0.904±0.003 vs 1.034±0.004,P0.05);HBcAg特异性DEXs刺激的PBMC产IFN-γ能力强于未成熟者(斑点数32±13 vs 14±2,P0.05),但也不如HBcAg特异性mDCs(32±13 vs 98±17,P≤0.01 )。 结论 HBcAg特异性mDEXs可有效激活自体PBMC增殖及分泌IFN-γ,其体外抗HBV特异性免疫活性强于未成熟者。
[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- 伪. 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- 纬 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 卤8.3% vs 16.5 卤6.8 vs 90.2 卤6.5% / 92.38 卤13.6% vs 6.3 卤4.5 卤4.5T = 83.53 卤10.8% vs 4.3 卤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 卤0.003 vs 0.545 卤0.010) P 0.001, but it was not as good as HBcAg specific mDCs(0.904 卤0.003 vs 1.034 卤0.004 P0.05G DEXs to stimulate PBMC to produce IFN- 纬. The number was 32 卤13 vs 14 卤2, P 0.05, but less than HBcAg specific mDCs(32 卤13 vs 98 卤17 (P 鈮，

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