Evaluation of the designed multi-epitope protein of Brucella melitensis in guinea pigs

Document Type : Original Article

Authors

1 Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

3 Veterinary Institution, Tehran, Iran

4 Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): One of the causes of human and animal zoonotic infections is Brucella melitensis, which is transmitted to humans through dairy products. It seems for prevention of human infection we might protect the livestock by an efficient protein as a vaccine candidate. For this purpose, the use of immunogenic proteins of bacteria is able to create immunity the same as the traditional vaccines.
Materials and Methods: In this study, by finding the immunogenic antigens of this bacterium by 2-dimensional gel electrophoresis and MALDI-TOF methods and also the proteins reported in other studies, we found the epitopes of the bacterial antigenic determinants in silico. Nineteen peptides of T and B epitopes were selected. They were ligated with linkers and after gene synthesis, the designed polypeptide was expressed in  Escherichia coli BL21. The purified recombinant MEL protein mixed with chitin was injected subcutaneously into three 300 g male guinea pigs three times. Also, PBS control and Rev.1 commercial vaccine groups were considered.
Results: The results show that MEL polypeptide is equal to the Rev.1 vaccine in stimulating secretion of IFNγ and IL2 and specific IgG. High levels of IL-2 emphasize the activation of the cellular immunity, and in particular comparison of PI in guinea pig’s spleen cells treated with recombinant MEL protein on days 0 and 5 show that it has significant proliferation compared with PBS unstimulated cells.
Conclusion: This recombinant protein could be a subunit protein with sufficient efficiency in stimulating the humoral and cellular-mediated immune system against B. melitansis.

Keywords

References

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Iranian Journal of Basic Medical Sciences
Volume 24, Issue 6
June 2021
Pages 833-841

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