A NOVEL MULTI-EPITOPE SUBUNIT VACCINE AGAINST BRUCELLA ABORTUS AND MELITENSIS

Background and Aim:Brucellosis is a zoonotic disease that can cause abortion in domestic animals and severe disease in humans. Brucellosisis exhibited as an endemic disease in iran. Live attenuated Brucella vaccines have several drawbacks. Peptide-based vaccines have been advocated as an attractive approach for prevention or treatment of infectious diseases. The reverse vaccinology has introduced new candidates for Brucella such as BhuA, FliC, 7α- HSDH through computer analysis.Methods:B cell and CD4+ and CD8+T cell epitopes from BhuA, FliC, 7α-HSDH antigens were selected and arranged in different patterns. The two and three-dimensional structure of the constructs were evaluated. Then, expression of the synthetic constructs were performed using pET28a expression vector in E. coli BL21(DE3). The proteins were purified with Ni-NTA column. SDS-PAGE and Western blot was used for confirmation of purified proteins. After expression of multi-epitopes, we evaluated mice humoral and cellular immune responses.Results:Antigenic regions of BhuA, FliC, 7α-HSDH were found and assembled in construct for stimulating humoral and cellular immunity. SDS-PAGE and Western blotting results indicated the similarity of in silico designing and in vitro expression. Vaccination of BALB/c mice with the recombinant proteins provided the significant protection level against B. melitensis and B. abortus challenge.Conclusion:In the present study, we designed the novel multi-epitope proteins from brucella antigens.The present study suggests that the peptide based vaccine as potential candidates for protection against Brucella.

Keywords:

Brucella , multi-epitope vaccine , new candidate , in-silico