Zhu Ai-hua, Jin Liang, Liu Jing-jing, Liu Mei-yan, Lv Ai-jun, Zheng Yuan-lin
School of Life Science, Xuzhou Normal University, Xuzhou, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 Nov;27(11):1165-8.
To study the efficacy of heat shock protein 65 kDa (HSP65) of Mybobacterium tuberculosis var. bovis in prevention of autoimmune diabetes by intranasal.
The HSP65 gene was derived from Mybobacterium tuberculosis var. bovis genome by PCR and successfully expressed as soluble protein in Escherichia coli. The recombinant protein HSP65 was purified by anion exchange column chromatography, then used to immunize prediabetic NOD (non-obese diabetic) mice via three intranasal (i.n.) delivery in absence of adjuvants. Serum samples from the immunized mice were collected at monthly intervals. The anti-HSP65 antibody was detected by enzyme-linked immunosorbent assay (ELISA) and verified by Western blot analysis. The concentration of blood glucose was measured by automatic analyzer.
Specific anti-HSP65 antibodies were successfully induced in mice immunized via intranasal routes. Histochemical analysis of mice pancreas tissue showed that HSP65 intranasal vaccination could decrease pathological changes in NOD mice.
Intranasal vaccination with HSP65 in NOD mice could prevent the development of diabetes. Our results demonstrate that intranasal vaccination with HSP65 reduces significantly the inflammatory process associated with auto-immune diabetes. This approach may offer novel therapeutic avenues for the treatment for of type 1 diabetes mellitus.
研究牛分枝杆菌65 kDa热休克蛋白（HSP65）经鼻内接种预防自身免疫性糖尿病的效果。
通过聚合酶链反应（PCR）从牛分枝杆菌基因组中获取HSP65基因，并在大肠杆菌中成功表达为可溶性蛋白。重组蛋白HSP65经阴离子交换柱层析纯化，然后在无佐剂的情况下通过三次鼻内（i.n.）给药用于免疫糖尿病前期非肥胖糖尿病（NOD）小鼠。每月收集免疫小鼠的血清样本。通过酶联免疫吸附测定（ELISA）检测抗HSP65抗体，并通过蛋白质印迹分析进行验证。用自动分析仪测量血糖浓度。
通过鼻内途径免疫的小鼠成功诱导出特异性抗HSP65抗体。小鼠胰腺组织的组织化学分析表明，HSP65鼻内接种可减少NOD小鼠的病理变化。
NOD小鼠鼻内接种HSP65可预防糖尿病的发生。我们的结果表明，HSP65鼻内接种可显著减轻与自身免疫性糖尿病相关的炎症过程。这种方法可能为1型糖尿病的治疗提供新的治疗途径。
