【摘要】 目的探讨基因重组腺病毒疫苗Ad-Aβ_（3-10）10-CpG鼻粘膜免疫APPswe/PSEN1d E9双转基因鼠诱导的炎症反应。方法 18只雄性10月龄APPswe/PSEN1dE9鼠,随机分为3组,分别以Aβ_3-10多价腺病毒疫苗AdAβ_（3-10）10-CpG、空腺病毒载体鼻粘膜免疫及Aβ_1-42肽肌内注射免疫,用MTT法检测脾细胞增殖反应、ELISA法检测脾细胞培养上清和脑组织匀浆中IL-4和INF-γ水平,免疫组化法检测脑内星形胶质细胞及淋巴细胞浸润。结果 Ad-Aβ_（3-10）10-CpG组和Aβ_1-42组,在相对应的免疫原刺激孔产生较高水平的增殖率,高于非相应免疫原刺激孔（P<0.05）,但低于Con A刺激孔（P<0.05）。皮质和海马区GFAP阳性细胞所占面积百分比为:空腺病毒载体组>Aβ_1-42组>Ad-Aβ_（3-10）10-CpG组。各组小鼠脑组织在血管内和脑实质偶尔发现个别CD3、CD5阳性细胞,3组没有显著差异。Ad-Aβ_（3-10）10-CpG组和Aβ_1-42组中在其相对应的免疫原刺激时检测到较高水平的IL-4、IFN-γ。Aβ_1-42组在Aβ_1-42肽刺激孔的IFN-γ水平显著高于Ad-Aβ_（3-10）10-CpG组在Aβ_3-10肽刺激孔的IFN-γ水平（P<0.05）。AdAβ_（3-10）10-CpG组脑组织匀浆IL-4水平大于Aβ_1-42组,但没有显著性差异（P>0.05）,Ad-Aβ_（3-10）10-CpG组脑组织匀浆中IFN-γ水平显著小于Aβ_1-42组（P<0.05）。结论腺病毒疫苗Ad-Aβ_（3-10）10-CpG鼻粘膜免疫APPswe/PSEN1dE9双转基因鼠主要产生Th2型免疫应答,可以减少脑内星形胶质细胞活化,未引起脑内炎症反应,即避免了Aβ_1-42全肽段所引起的炎症反应。更多还原

【Abstract】 Objective To study the inflammatory response after intranasal inoculation with a recombinant adenovirus vaccine Ad-Aβ_（3-10）10-CpG in APPswe/PSEN1dE9 transgenic mice. Methods Eighteen ten-month old male APPswe/PSEN1dE9 mice were divided into three groups and immunized with recombinant adenovirus vaccine Ad-Aβ_（3-10）10-CpG,empty adenoviral vector and Aβ_1-42 peptide,representatively. Spleen T-cell proliferation response was determined with MTT assay. Levels of IL-4 and INF-γ in spleen cell culture supernatant and brain homogenate were determined with ELISA. Astrocytes and CD3,CD5 and CD45 positive cells in the brain were detected with immunohistochemistry. Results Ad-Aβ_（3-10）10-CpG group and Aβ_1-42 peptide group showed higher levels of proliferation rate restimulated with its corresponding antigen（ P < 0. 05）,but lower than the Con A-stimulated wells（ P < 0. 05）. The percentage of area of GFAP positive cells in cortex and hippocampus was as follows: empty vector group > Aβ_1-42 peptide group > Ad-Aβ_（3-10）10-CpG group. A very few CD5-positive and CD3-positive lymphocytes were occasionally observed within the blood vessels or in the parachyma in all three groups of mice. There were no significant different in all three groups. Ad-Aβ_（3-10）10-CpG and Aβ_1-42 peptide group exhibited significantly greater IL-4 and IFN-γ levels in the cultures of spleen T cells restimulated with its corresponding antigen. Mice immunized with Aβ_1-42 peptide exhibited the greater IFN-γ levels in spleen T cells restimulated with Aβ_1-42 peptide than mice immunized with Ad-Aβ_（3-10）10-CpG（ restimulated with Aβ3-10）（ P < 0. 05）. IL-4 levels in brain homogenates from mice of Ad-Aβ_（3-10）10-CpG group were greater than that of Aβ_1-42 peptide group,but the diffrence was not significant（ P> 0. 05）. IFN-γ levels in brain homogenates of Aβ_1-42 peptide group was greater than that of Ad-Aβ_（3-10）10-CpG group（ P <0. 05）. Conclusion Intranasal inoculation with a recombinant adenovirus vaccine Ad-Aβ_（3-10）10-CpG appeared to induce a Th2 type immune response and reduction of astrocytosis without causing neuroinflammation in the brain. Ad-Aβ_（3-10）10-CpG had low potential to cause autoimmune response caused by Aβ_1-42 peptide.更多还原