Aβ_(3-10)多价腺病毒疫苗鼻粘膜免疫AD转基因鼠的治疗效果研究  

The therapeutic effect of multivalent adenovirus vaccine Aβ_(3-10)after intranasal inoculation in AD transgenic mice

作　　者：李昱[1] 张慧[1] 林璐璐[1] 辛世萌[1] 曹云鹏[2] LI Yu ZHANG Hui LIN Lulu et al(Department of Neurology, The Second Affiliated Hospital of Dalian Medical University, Dalian 116027, China)

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1期4-8,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Aβ_(3-10)多价腺病毒疫苗鼻粘膜免疫AD转基因鼠的治疗效果。方法 18只雄性10月龄AD转基因鼠,随机分为3组,分别以Aβ_(3-10)多价腺病毒疫苗Ad-Aβ_((3-10)10)-Cp G、空腺病毒载体及Aβ1-42免疫,ELISA法检测血清抗Aβ抗体滴度及亚型,Morris水迷宫检测转基因鼠的学习记忆能力,免疫组化法检测转基因鼠脑内Aβ沉积;ELISA法检测转基因鼠脑组织匀浆和血清中可溶性Aβ42水平。结果 Ad-Aβ_((3-10)10)-Cp G组和Aβ1-42组抗体水平随着免疫次数逐渐增加,第7次免疫后Ad-Aβ_((3-10)10)-Cp G组和Aβ1-42组血清中抗Aβ抗体水平分别为(67.42±13.68)μg/ml和(94.41±14.01)μg/ml,而空载体组一直在基线水平。Ad-Aβ_((3-10)10)-Cp G组Ig G1/Ig G2a比值明显高于Aβ1-42组(P<0.05)。在Morris水迷宫实验中Ad-Aβ_((3-10)10)-Cp G组的逃避潜伏期明显小于空载体组(P<0.01);Ad-Aβ_((3-10)10)-Cp G组在靶象限的停留时间明显长于空载体组(P<0.01);Ad-Aβ_((3-10)10)-Cp G组穿越平台所在位置的次数明显多于空载体组(P<0.05)。Ad-Aβ_((3-10)10)-Cp G组脑组织Aβ沉积所占面积百分比与空载体组比较明显减少(P<0.01)。Ad-Aβ_((3-10)10)-Cp G组脑组织匀浆和血清中可溶性Aβ42水平明显高于空载体组(P<0.01)。结论 Aβ_(3-10)多价腺病毒疫苗鼻粘膜免疫AD转基因鼠,主要引起Th2型免疫应答,可以改善AD转基因鼠学习和记忆能力,促进转基因鼠脑内Aβ清除,可以减少由细胞免疫应答引起的炎症反应。Aβ_(3-10)多价腺病毒疫苗是AD免疫治疗的安全有效的候选疫苗。Objective To study the therapeutic effects of multivalent adenovirus vaccine Aβ_（3-10） after intranasal inoculation in AD transgenic mice. Methods Eighteen ten-month old male transgenic mice were divided into three groups randomly and immunized with recombinant adenovirus vaccine Ad-Aβ（ 3-10） 10-CpG,empty adenoviral vector and Aβ_（1-42） peptide,representatively. Titers of anti-Aβ antibody and isotypes of immunoglobin in sera were determined with ELISA. Cognitive function of the transgenic mice was detected by Morris water maze tests. Aβ burden in brains of the mice was detected with Immunohistochemistry. Soluble Aβ_（42）in plasma and brain homogenate was detected with ELISA. Results Sera from mice vaccinated with Ad-Aβ（ 3-10） 10-CpG and Aβ_（1-42） showed a steady increase in anti-Aβ antibody titer with each boost,reaching an average of（ 67. 42 ± 13. 68） μg / ml and（ 93. 41 ± 14. 01） μg / ml after the final immunization,respectively. Antibody titers from mice vaccinated with empty vector remained at background level. Ig G1 / Ig G2 a ratio of the Ad-Aβ（ 3-10） 10-CpG group was much greater than that of the Aβ_（1-42） group（ P〈0. 05）. Morris water maze tests： Ad-Aβ（ 3-10） 10-CpG immunized mice showed shorter escape latency compared with the empty vector group（ P〈0. 01）. Ad-Aβ（ 3-10） 10-CpG immunized mice spent significant more time in the target quadrant than empty vector immunized mice（ P〈0. 01）,Ad-Aβ（ 3-10） 10-CpG immunized mice crossed the platform location significantly more often than empty vector immunized mice（ P〈0. 05）. Percentage of Aβ burden occupied area significantly reduced in mice immunized with Ad-Aβ（ 3-10） 10-CpG compared with empty vector group. Soluble Aβ42levels significantly increased in the brain homogenates and plasma of the Ad-Aβ（ 3-10） 10-CpG immunized mice compared with empty vector immunized mice（ P〈0. 01）. Conclusions Intranasal inoculation with a recombinant adenovirus vaccine Ad-

关 键 词：阿尔茨海默病 腺病毒疫苗 AΒ 免疫治疗 

分 类 号：R749.16[医药卫生—神经病学与精神病学]