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中国临床药理学与治疗学 ›› 2006, Vol. 11 ›› Issue (8): 857-862.

• 研究原著 • 上一篇    下一篇

鼻粘膜免疫融合蛋白Hsp65-6 ×p277预防NOD 小鼠1 型糖尿病的发生

金亮, 王宇, 朱爱华, 刘景晶   

  1. 中国药科大学生命科学与技术学院, 南京 210009, 江苏
  • 收稿日期:2006-05-30 接受日期:2006-06-26 出版日期:2006-08-26 发布日期:2020-11-05

Intranasal vaccination with p277 tandem repeat sequences carried by Hsp65 prevented type 1 diabetes in NOD mice

JIN Liang, WANG Yu, ZHU Ai-hua, LIU Jing-jing   

  1. Minigene Pharmacy Laboratory , School of Life Science &Technology ,China Pharmaceutical University , Nanjing 210009 , Jiangsu, China
  • Received:2006-05-30 Accepted:2006-06-26 Online:2006-08-26 Published:2020-11-05
  • About author:JIN Liang, male, doctor, majoring in gene engineering medicine.Tel:13852289934 E-mail:jinboshi1975@yahoo.com.cn
  • Supported by:
    National High Technology “863”Programs of China(No2002AA217031-2);National Natural Science Foundation of China(No30270298);Natural Science Foundation of Jiangsu (NoBK95092309;NoBG2001011)

摘要: 目的 提高多肽p277 的免疫原性, 从而提高其对自身免疫性糖尿病的预防作用。方法 将p2776 次重复与HsP65 融合置于pET28a 中构建重组Hsp65-6 ×p277 表达质粒。该重组质粒在大肠杆菌BL21 中以高效可溶形式表达。依次通过细胞裂解、硫酸铵沉淀、双蒸水透析、DEAE 纤维素52 柱层析纯化获得目的蛋白。用纯化后的融合蛋白HsP65-6 ×p277 通过鼻腔给药方式, 在不添加任何佐剂的情况下3 次免疫4 周龄雌性NOD 小鼠。每月眼角取血,检测抗体和血糖浓度。结果 初步药效学实验表明融合蛋白HsP65-6 ×p277 可抑制NOD 小鼠中1 型糖尿病的发生。结论 融合蛋白Hsp65-6 ×p277 有可能发展成为一种具有防治胰岛素依赖性糖尿病作用的疫苗。

关键词: 热休克蛋白65, p277, 胰岛素依赖性糖尿病, 免疫

Abstract: AIM: To improve the prevent efficacy of peptide p277 in autoimmune diabetes.METHODS: The recombinant expression plasmid pET28-Hsp65-6×p277 was constructed by inserting 6×p277 which were amplified by PCR into the vector pET28-Hsp65.The plasmid pET28-Hsp65-6×p277 was transformed into E.coli BL21 (DE3) and the fusion protein (Hsp65-6×p277) was expressed effectively as soluble protein after inducing by lactose.The fusion protein was purified and then used to immunize 4-week old female NOD mice with three times of i.n.inoculations in the absence of adjuvants.Serum samples from the immunized mice were collected at monthly interval.The concentrations of blood glucose and antibodies were measured by automatic analyzer.RESULTS: Administration with the Hsp65-6×p277 to NOD mice could prevent the development of diabetes.CONCLUSION: The fusion protein Hsp65-6×p277 might be further developed to a vaccine against insulin-dependent diabetes mellitus.

Key words: heat shock protein 65, p277, insulindependent diabetes mellitus, immune

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