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Chinese Journal of Clinical Pharmacology and Therapeutics ›› 2006, Vol. 11 ›› Issue (8): 857-862.

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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)

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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