Aim: To study the efficacy of heat shock protein 65 kDa (HSP65) of Mybobacterium tuberculosis var. bovis in prevention of autoimmune diabetes by intranasal.
Methods: 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.
Results: 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.
Conclusion: 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.
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