中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (27): 2581-2590.doi: 10.3969/j.issn.1673-5374.2013.27.010

• 原著:退行性病与再生 • 上一篇    

一种融合C3d-p28的新型DNA疫苗诱导了C57BL/6J小鼠针对β-淀粉样蛋白的Th2免疫反应

  

  • 收稿日期:2013-05-05 修回日期:2013-08-02 出版日期:2013-09-25 发布日期:2013-09-25
  • 基金资助:

    国家自然科学基金（No. 30471927）

A new DNA vaccine fused with the C3d-p28 induces a Th2 immune response against amyloid-beta

Wanshu Guo1, Sha Sha1, Tongzi Jiang2, Xiaona Xing1, Yunpeng Cao1   

  1. 1 Department of Neurology, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China

    2 Department of Neurology, First People’s Hospital of Shenyang City, Shenyang 110041, Liaoning Province, China
  • Received:2013-05-05 Revised:2013-08-02 Online:2013-09-25 Published:2013-09-25
  • Contact: Yunpeng Cao, M.D., Chief physician, Doctoral supervisor, Department of Neurology, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China, ypengcao@ yahoo.com.
  • About author:This study was supported by the National Natural Science Foundation of China, No. 30471927.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 30471927.

PDF

1559

摘要：

阿尔茨海默病Aβ主动免疫可以增加Aβ的清除率，减轻进行性认知能力减退，但易产生脑膜脑炎的不良反应，而这可能与Th1淋巴细胞的激活有关。为了增加抗淀粉样蛋白-β（Aβ）抗体生成，诱导Th2免疫反应来避免不良反应，我们构建了新的DNA疫苗p(Aβ3-10)10- C3d-p28.3，编码10倍的Aβ3-10和3倍的C3d-p28作为分子佐剂，肌肉注射8-10周龄的C57BL/6J鼠。ELISA法检测血清中抗Aβ抗体的滴度、分型、脾T细胞细胞因子，MTT法检测脾T细胞增殖率。用12月龄的APP/PS1转基因鼠检测抗Aβ抗体与Aβ斑结合情况。结果DNA疫苗p(Aβ3-10)10- C3d-p28.3诱发了高滴度的抗Aβ抗体产生，并且产生的抗Aβ抗体能够与12月龄的APP/PS1转基因鼠的脑切片中Aβ斑结合，显示了该疫苗对阿尔茨海默病模型鼠是有效的。同时，该疫苗主要产生了IgG1体液反应，体外脾细胞培养中IFN-γ低水平，这说明了倾向于Th2型细胞免疫反应，避免了不良反应的发生。我们的结果表明，DNA疫苗p(Aβ3-10)10- C3d-p28.3是阿尔茨海默病Aβ免疫非常有希望的治疗措施。

 

关键词: 神经再生, 阿尔茨海默病, β-淀粉样蛋白, C57BL/6J小鼠, DNA疫苗, 主动免疫, 被动免疫, C3d-P28分子佐剂, Th2免疫反应, 基金资助文章

Abstract:

To enhance anti-amyloid-beta (Aβ) antibody generation and induce a Th2 immune response, we constructed a new DNA vaccine p(Aβ3–10)10-C3d-p28.3 encoding ten repeats of Aβ3–10 and three copies of C3d-p28 as a molecular adjuvant. In this study, we administered this adjuvant intramus-cularly to female C57BL/6J mice at 8–10 weeks of age. Enzyme linked immunosorbent assay was used to detect the titer of serum anti-Aβ antibody, isotypes, and cytokines in splenic T cells. A 3- (4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was used to detect the prolifera-tion rate of splenic T cells. Brain sections from a 12-month-old APP/PS1 transgenic mouse were used for detecting the binding capacities of anti-Aβ antibodies to Aβ plaques. The p(Aβ3–10)10-C3d-p28.3 vaccine induced high titers of anti-amyloid-β antibodies, which bound to Aβ plaques in APP/PS1 transgenic mouse brain tissue, demonstrating that the vaccine is effective against plaques in a mouse model of Alzheimer’s disease. Moreover, the vaccine elicited a pre-dominantly IgG1 humoral response and low levels of interferon-γ in ex vivo cultured splenocytes, indicating that the vaccine could shift the cellular immune response towards a Th2 phenotype. This indicated that the vaccine did not elicit a detrimental immune response and had a favorable safety profile. Our results indicate that the p(Aβ3–10)10-C3d-p28.3 vaccine is a promising immunothera-peutic option for Aβ vaccination in Alzheimer’s disease.

Key words: neural regeneration, Alzheimer’s disease, amyloid-β, C57BL/6J mice, DNA vaccine, active immunotherapy, passive immunotherapy, C3d-p28 molecular adjuvant, Th2 immune response, grants-supported paper, neuroregeneration