上海交通大学学报(医学版) ›› 2017, Vol. 37 ›› Issue (11): 1464-.doi: 10.3969/j.issn.1674-8115.2017.11.003

• 论著(基础研究) • 上一篇    下一篇

胆固醇酯转移蛋白纳米疫苗抗动脉粥样硬化的初步实验研究#br#

尤纱纱 1,李永勇 2,董海青 2,何斌 1   

  1. 1. 上海交通大学 医学院附属新华医院麻醉与重症医学科,上海 200092;2. 同济大学 医学院生物医学工程与纳米科学研究院,上海 200092
  • 出版日期:2017-11-28 发布日期:2018-01-10
  • 通讯作者: 何斌,电子信箱:hebin@xinhuamed.com.cn
  • 作者简介:尤纱纱(1992—),女,硕士生;电子信箱:yokia_youshasha@163.com
  • 基金资助:
    国家自然科学基金(81470390);上海市教育委员会高峰高原学科建设计划(20152218);上海申康医院发展中心促进市级医院临床技能与临床创新三年行 动计划(16CR3006A);上海市科学技术委员会科研计划项目(17411954700)

Study of a nanovaccine targeting cholesteryl ester transfer protein against atherosclerosis

YOU Sha-sha1, LI Yong-yong2, DONG Hai-qin2, HE Bin1   

  1. 1. Department of Anesthesiology and SICU, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China; 2. Department of Biomedical Engineering &Nano Science, Tongji University School of Medicine, Shanghai 200092, China
  • Online:2017-11-28 Published:2018-01-10
  • Supported by:
     National Natural Science Foundation of China, 81470390; Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support, 20152218; Clinical Research Plan of Shanghai Hospital Development Center, 16CR3006A; Program of Science and Technology Commission of Shanghai Municipality, 17411954700

摘要: 目的 · 评估胆固醇酯转移蛋白(CETP)纳米疫苗是否能提高CETP 半抗原免疫原性,并初步探讨其抗兔动脉粥样硬化的可 行性。方法 · 采用Fmoc 法合成巯基修饰的CETP 多肽;用交联剂Sulfo-SMCC 将 CETP 多肽分别与载体蛋白——卵清蛋白(OVA) 分子、OVA 纳米颗粒链接;免疫新西兰大白兔,检测实验兔血清的抗体、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇 (LDL-C)的浓度。设 PBS 对照组(n=3)、传统疫苗组(n=3)、纳米疫苗组(n=3),进行各组间比较。结果 · 动态光散射粒度仪结果 显示纳米疫苗粒径约 70 nm, zeta 电位约 -8.81 mV,透射电子显微镜观察结果显示纳米疫苗近似球形;紫外分光光度计检测结果显示 终产物最大吸收峰发生偏移,SDS-PAGE 结果显示终产物相对分子质量增加,皆证明抗原负载成功; ELISA 结果显示自第 3 周开始纳 米疫苗组的 D(450 nm)明显高于传统疫苗组。结论 · 该纳米疫苗可以提高 CETP 半抗原的免疫原性,激发实验兔产生更高的抗体。

关键词: 纳米, 疫苗, 胆固醇酯转移蛋白, 动脉粥样硬化

Abstract: Objective · To prepare nanocarriers capable of improving the immunogenicity of cholesteryl ester transfer protein (CETP) hapten.  Methods · Prepare CETP polypeptide modified by thiol in Fmoc, then Sulfo-SMCC were used to prepare CETP peptide-ovalbumin (OVA) nanoparticle and CETP peptide - OVA molecule conjugates in a two-step reaction scheme. The particle size and zeta potential of nanovaccine were determined and the morphology was observed. New Zealand White rabbits were vaccinated by subcutaneous injection of vaccine and serum collected from rabbits was detected by ELISA assay for the analysis of antibodies, high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol (LDL-C). The rabbits were randomly allocated to the PBS group (n=3), traditional vaccine group (n=3), and nanovaccine group (n=3).  Results · Nanovaccine targeting CETP were successfully synthesized, with about 70 nm in size and about -8.81 mV in zeta potential, and possessed homogeneous spherical shape under transmission electron microscopy. Compared with traditional vaccine group, rabbits in nanovaccine group got higher antibodies.  Conclusion · Nanovaccine improve immunogenicity of CETP haptens, and stimulate experiment rabbits to produce higher antibodies.

Key words: nano, vaccine, cholesteryl ester transfer protein, atherosclerosis