CFP10: mFcγ2 as a novel tuberculosis vaccine candidate increases immune response in mouse

Document Type: Original Article

Authors

1 Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 HIV/AIDS, HTLV and Viral Hepatitis Research Center, Iranian Academic Center for Education, Culture and Research (ACECR), Mashhad, Iran

4 Immunology Research Center, Inflammation and Inflammatory Diseases Division, Medical school, Mashhad University of Medical Sciences, Mashhad, Iran

5 Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran

6 Organic Chemistry, Department of Laboratory Sciences, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Despite treatment with antibiotics and vaccination with BCG, tuberculosis (TB) is still considered as one of the most important public health problems in the world. Therefore, designing and producing a more effective vaccine against TB seems urgently. In this study, immunogenicity of a fusion protein which consisting or comprising CFP-10 from Mycobacterium tuberculosis and the Fc-domain of  mouse IgG2a was evaluated as a novel subunit vaccine candidate against TB.
Materials and Methods: The genetic constructs were cloned in pPICZαA expression vector and recombinant vectors (pPICZαA-CFP-10: Fcγ2a and pPICZαA-CFP-10:His) were transformed into Pichia pastoris. To evaluate the expression of recombinant proteins, SDS-PAGE and immunoblotting were used. The immunogenicity of recombinant proteins, with and without BCG were assessed in BALB/c mice and specific cytokines against recombinant proteins (IFN-γ, IL-12, IL-4, IL-17 and TGF-β) were evaluated.
Results: The levels of IFN-γ and IL-12 in mice that received recombinant proteins was higher than the control groups (BCG and PBS). Thus, both recombinant proteins (CFP-10:Fcγ2a and CFP-10:His) could excite good response in Th1-cells. The Fc-tagged protein had a stronger Th1 response with low levels of IL-4, as compared to CFP-10:His. However, the highest level of Th1 response was observed in groups that were vaccinated with BCG (prime) and then received recombinant protein CFP-10: Fcγ2a (booster).
Conclusion: The results demonstrated that binding mice Fc-domain to CFP-10 protein can increase the immunogenicity of the subunit vaccine. Further studies, might be able to design and produce a new generation of subunit vaccines based on the Fc-fused immunogen.

Keywords


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