Investigating the effect of Alginate acid nanoparticles and montanide and freond adjuvants the efficacy of a recombinant vaccine candidate against Shigella

Shigellosis, an acute intestinal infection, is one of the global health problems caused by Shigella pathogenic species in humans. Production of a cost-effective and protective vaccine against pathogenic species of this bacterium can have a significant effect on public health. The purpose of this research is to express a recombinant chimeric protein as a recombinant immunogen candidate against Shigella pathogenic species and to investigate its immunogenicity with Freund and Montanide adjuvants and protein loaded in allergenic nanoparticles. For this purpose, the gene encoding chimera protein was subhomogenized in pET۳۲α(+) vector. The correctness of the target gene subisolation was confirmed using PCR reaction. The recombinant vector pET۳۲α(+) containing the desired gene fragment was transferred into the expression host E.coli BL۲۱ (DE۳) and the expression of the recombinant protein was induced using IPTG. The target was purified by urea gradient method. Western blotting method was used to confirm the expression of the recombinant protein. Finally, the purified protein was administered orally and subcutaneously to mice in four groups (combined with montanide adjuvant, Freund's adjuvant, loaded in allergenate gel and control group) on four occasions. Antibody titer against recombinant protein with Montanide and Freund adjuvants and allergenate gel in the serum of mice immunized subcutaneously was ۱/۳۲۰۰ and immunized orally by allergenate gel loaded with recombinant protein was ۱/۱۶۰۰. Finally, the evaluation of Shigella flexneri neutralization test using antibody produced by immunized animals showed that the serum of immunized animals is capable of inactivating Shigella flexneri. In general, the results of this research show that this recombinant protein is multi-epitope due to having the main epitopes of common antigens between Shigella pathogenic species and also due to the appropriate titer produced against this protein in combination with adjuvants. Freund and Montanide and loading allergens in the gel and the neutralizing power of the resulting antibodies can be a suitable vaccine candidate for the prevention of shigellosis.

کلیدواژه ها:

Shigella ، shiglose ، recombinant protein ، multi-VP vaccine ، nanoparticles ، adjuvant