Aβ3-10s基因疫苗免疫AD小鼠诱导Th2型免疫反应的研究  

An vaccine Aβ3-10 induces Th2 immune response in AD mice

作　　者：郭蓉[1] 姜童子[2] 黄葵[3] 李昱[2] 陈晓虹[1] 曹云鹏[2] 

机构地区：[1]辽宁省人民医院神经内科,辽宁沈阳110016 [2]中国医科大学附属第一医院神经内科,辽宁沈阳110001 [3]沈阳市红十字会医院干诊科,辽宁沈阳110013

出　　处：《中风与神经疾病杂志》2013年第5期388-391,共4页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金课题(30471927)

摘　　要：目的探讨新型AD疫苗AdCpG-(Aβ3-10)10诱导转基因小鼠产生的免疫反应类型。方法将人工合成的Aβ3-10的基因片段插入到真核表达质粒pcDNA3.1+,以CpG为基因佐剂,使用E1/E3缺陷的腺病毒载体pAxCAwtit作为基因载体,获得基因重组缺陷腺病毒AdCpG-(Aβ3-10)10。以3月龄的转基因小鼠18只为研究对象,实验分为3组,每组各6只小鼠:免疫含有目的基因的AdCpG-(Aβ3-10)10组、不含目的基因的AdCpG组及Aβ1-42全肽链组。各组均经鼻粘膜分别免疫AdCpG-(Aβ3-10)10、不含目的基因的AdCpG及Aβ1-42全肽链,每组均免疫8次,每3个月1次。用ELISA方法检测小鼠血清中IgG抗体含量及抗体分型,用MTT方法观察脾细胞增值反应;用ELISA法检测IL-2、IL-4、IL-10及γ-干扰素等细胞因子含量。结果小鼠经过6次免疫后,AdCpG-(Aβ3-10)10组和Aβ1-42组抗Aβ42 IgG抗体浓度分别为21.17±1.59ng/ml、23.93±1.66ng/ml,较AdCpG组(1.08±0.16)明显升高(P<0.05),但AdCpG-(Aβ3-10)10组的IgG1/IgG2a(12.94±3.79)较Aβ1-42组的IgG1/IgG2a(6.13±2.21)明显增高(P<0.01);AdCpG-(Aβ3-10)10组和Aβ1-42组的脾细胞在体外培养,经Con A刺激后细胞增殖明显,OD值分别为0.37±0.059,0.3237±0.048与AdCpG(0.1692±0.072)比较有明显差异(P<0.05)。经AdCpG-(Aβ3-10)10免疫的小鼠脾细胞体外培养液中IL-4和IL-10含量分别为342.34±45.68μg/ml、329.46±112.99μg/ml,经Aβ1-42免疫的小鼠脾细胞体外培养液中IL-2、INF-γ、IL-4和IL-10含量分别为284.81±22.55μg/ml、361.96±26.47μg/ml、223.19±22.45μg/ml、258.68±102.42μg/ml,与AdCpG(149.03±27.34μg/ml、291.92±45.92μg/ml、169.52±25.68μg/ml、134.85±21.67μg/ml)比较有明显差异(P<0.05)。结论 AdCpG-(Aβ3-10)10主动免疫幼龄双转基因小鼠主要产生Th2型体液免疫应答反应,可以减少细胞免疫应答反应引起的炎性反应。Objective To Study the immune reaction type after immune transgene mice with the vaccine,determined the serum IgG antibodies and the cytokines of Spleen cell proliferation.Methods Synthesized Aβ3-10 gene was cloned into the eukaryotic expression plasmid,with CpG as adjuvant,the use of E1/E3 defective adenovirus vector as a gene carrier,obtained recombinant adenovirus vaccine AdCpG-（Aβ3-10）10.Immuned double transgenic mice B6.Cg-Tg（APPswe,PSEN1dE9） by nasal with the vaccine.Set the three experimental groups,the target gene AdCpG-（Aβ3-10） 10,non-target gene AdCpG group,Aβ1-42 peptide group.Then the whole mice were immunized once every three weeks and during the eight times immunations the blood serum were acquaired.Finaly,examed the IgG antibodies by ELISA and value-added Con A stimulated spleen cells and their antigen Stimulation results after stimulation inflammatory cytokines such as IL-2,IL-4,IL-10 and γ-interferon levels to identify the type of immune response.Results After the sixth immunization AdCpG-（Aβ3-10）10group and Aβ1-42 group of mice at high concentrations of antibody detection,there were significant difference with the control group.The IgG1/IgG2a of AdCpG-（Aβ3-10）10group was higher than Aβ1-42 group.The spleen cells in vitro by antigen or Con A physical stimulation increased significantly compared with the control group.Respectively the IL-4 and IL-10 of AdCpG-（Aβ3-10）10 group were higher than the control group,while cytokines IL-2,INF-γ,IL-4 and IL-10 of Aβ42 group were all higher compared with the control group.Conclusion The active immunation of AdCpG-（Aβ3-10）10 mainly produced Th2 type humoral immune response,and reduced inflammatory reaction by the cellular immune response.

关 键 词：阿尔茨海默病 Aβ疫苗 免疫反应 

分 类 号：R749.16[医药卫生—神经病学与精神病学]