中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (3): 519-527.doi: 10.4103/1673-5374.266061

• 原著:退行性病与再生 • 上一篇    下一篇

早期主动Aβ疫苗注射可减少小鼠海马tau蛋白磷酸化保护认知功能

  

  • 出版日期:2020-03-15 发布日期:2020-05-26
  • 基金资助:

    中国国际自然科学基金项目(81371227

Early active immunization with Aβ3–10-KLH vaccine reduces tau phosphorylation in the hippocampus and protects cognition of mice

Jin-Chun Wang1, Kun Zhu2, Hui-Yi Zhang2, Guo-Qing Wang2, Hui-Ying Liu1, Yun-Peng Cao2   

  1. 1 Department of Neurology, the Fifth People’s Hospital of Shenyang, Shenyang, Liaoning Province, China
    2 Department of Neurology, the First Hospital of China Medical University, Shenyang, Liaoning Province, China
  • Online:2020-03-15 Published:2020-05-26
  • Contact: Yun-Peng Cao, MD,yun_pengcao@sina.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81371227 (to YPC).

摘要:

主动和被动抗Aβ免疫疗法已成功用于预防和治疗阿尔茨海默病的动物模型中,然而临床上使用这些免疫疗法效果不佳,可能是因为疫苗接种开始较晚。因此,实验设计:(1)APP/PS1/tau转基因小鼠模型1月龄起在颈部皮下注射100 μL Aβ3-10-KLH肽(疫苗)(2μg/μL),然后于1.52.53.54.55.56.5月龄时再次注射Aβ3-10-KLH肽;然后以ELISA检测血清中的Aβ抗体水平,以Morris水迷宫评估空间学习记忆能力,以免疫组化检测海马组织中HT7、磷酸盐磷光体依赖性AT8及AT180免疫阳性反应,以Western blot分析海马组织中AT8和AT180表达水平;(2)测试结果发现,注射疫苗后,小鼠产生高水平的Aβ抗体,认知功能明显改善,总tau和磷酸化tau水平明显降低。表明早期Aβ3-10-KLH疫苗主动注射免疫,可通过抑制tau磷酸化,减轻3xTg-AD小鼠的认知衰退。实验于201642日经中国,沈阳,中国医科大学动物伦理委员会批准(批准号103-316)。


orcid: 0000-0002-5933-2590 (Yun-Peng Cao)

关键词: 阿尔茨海默病, 淀粉样蛋白β, Aβ3-10-KLH疫苗, tau蛋白磷酸化, 认知能力下降, 3×Tg-AD小鼠, 淀粉样蛋白前体蛋白, 转基因小鼠, 神经再生

Abstract: Active and passive anti-Aβ immunotherapies have successfully been used for the prevention and treatment of Alzheimer’s disease animal models. However, clinical use of these immunotherapies is not effective, because the vaccination is administered too late. At 1 month of age, 100 μL of Aβ3–10-KLH peptide (vaccine, 2 μg/μL) was subcutaneously injected into the neck of an amyloid precursor protein/presenilin- 1/tau transgenic (3×Tg-AD) mouse model. Aβ3–10-KLH peptide was re-injected at 1.5, 2.5, 3.5, 4.5, 5.5, and 6.5 months of age. Serum levels of Aβ antibody were detected by enzyme-linked immunosorbent assay, while spatial learning and memory ability were evaluated by Morris water maze. Immunohistochemistry was used to detect total tau with HT7 and phosphorylated tau with AT8 (phosphorylation sites Ser202 and Thr205) and AT180 (phosphorylation site Thr231) antibodies in the hippocampus. In addition, western blot analysis was used to quantify AT8 and AT180 expression in the hippocampus. The results showed that after vaccine injection, mice produced high levels of Aβ antibody, cognitive function was significantly improved, and total tau and phosphorylated tau levels were significantly reduced. These findings suggest that early active immunization with Aβ3–10-KLH vaccine can greatly reduce tau phosphorylation, thereby mitigating the cognitive decline of 3×Tg-AD mice. This study was approved by the Animal Ethics Committee of China Medical University, China (approval No. 103-316) on April 2, 2016.

Key words: 3×Tg-AD, Aβ3–10-KLH vaccine, Alzheimer’s disease, amyloid precursor protein, amyloid-beta, cognitive decline, tau phosphorylation, transgenic mouse