阿尔茨海默病核酸疫苗的构建及诱导体液免疫反应的初步研究  

Induction of humoral immune response in BALB/c mice by plasmid DNA vaccines against Alzheimer's disease

作　　者：张助[1,2] 赵志虎[1] 朱玲玲[3] 赵彤[3] 范明[3] 马清钧[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]解放军464医院,天津300381 [3]军事医学科学院基础医学研究所,北京100850

出　　处：《生物技术通讯》2003年第6期484-488,498,共6页Letters in Biotechnology

摘　　要：近年,内源性蛋白主动免疫预防和治疗阿尔茨海默病(AD)的策略成为AD研究领域的热点,并获得迅猛发展。构建了以β-片层结构的淀粉样蛋白(Aβ)及其可溶性与病理性突变体为抗原基因的DNA疫苗用于免疫小鼠,经过对免疫方案的探索和调整,结果能够打破其自身耐受,在外周血中诱发出较高滴度的抗-Aβ抗体。初步探讨了诱导体液免疫学效应的规律。并在原代培养的海马神经元Aβ毒性模型中验证了免疫后血清的生物学活性。AD peptide vaccines were tested in clinical trials after a great success in transgenic mouse model of AD. Data indicates that and-Aβ antibodies play an important role in removing of brain amyloid depositions either in AD mice or in patients. To elicit appropriate and long lasting humoral response, we tried a different method of immunization,DNA vaccine. Sequences encoding one wild-type and another two mutants of Aβ genes were similarly cloned into pcDNA3.1hisA(-) for mammalian cell expression. BALB/c mice were immunized by intramuscular(i.m.) injections of 200 μg of plasmids per mouse every time without adjuvant. All animals received a total of 5-6 inoculations over a 3-4.5 month period(depending on different immunization protocols). Anti-Aβ antibody titers(IgG) were detected by enzyme-linked immunoadsordent assay. Strong humoral response were elicited in two of three groups, with average titers up to 1:11500 and 1:55600 to Ap. Average antibody titer of the third group also reached to 1:2640. More plasmids and frequent inoculations could activate first response or break the self-tolerance of mice. Anti-serums were also tested for their biological function to prevent cultured rat hippocampal neurons from the loxicily of Aβ fragmenls.

关 键 词：阿尔茨海默病 淀粉样蛋白 核酸疫苗 体液免疫反应 主动免疫 

分 类 号：R749.16[医药卫生—神经病学与精神病学] Q78[医药卫生—临床医学]