Aβ_(3-10)多价腺病毒疫苗鼻粘膜免疫阿尔茨海默病转基因鼠的炎症反应研究  

Inflammatory response after intranasal inoculation with an adenovirus vaccine encoding multivalent Aβ_(3-10) in Alzheimer's disease transgenic mice

作　　者：李昱[1] 宋贵军[1] 林璐璐[1] 辛世萌[1] 曹云鹏[2] 

机构地区：[1]大连医科大学附属第二医院神经内科,辽宁大连116027 [2]中国医科大学附属第一医院神经内科,辽宁沈阳110001

出　　处：《中风与神经疾病杂志》2017年第5期393-397,共5页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金(No.81371227)

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。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 A

关 键 词：阿尔茨海默病 淀粉样蛋白 炎症反应 星形胶质细胞 

分 类 号：R749.1[医药卫生—神经病学与精神病学]