Author/Authors :
Baghani، Ali Asghar نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,Mashhad University of Medical Sciences,Mashhad,Iran , , Soleimanpour، Saman نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,Mashhad University of Medical Sciences,Mashhad,Iran , , Farsiani، Hadi نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,Mashhad University of Medical Sciences,Mashhad,Iran , , Mosavat، Arman نويسنده HIV/AIDS, HTLV and Viral Hepatitis Research Center,Iranian Academic Center for Education, Culture and Research (ACECR),Mashhad,Iran , , Yousefi، Masoud نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,Mashhad University of Medical Sciences,Mashhad,Iran , , Meshkat، Zahra نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,Mashhad University of Medical Sciences,Mashhad,Iran , , Rezaee، Abdolrahim نويسنده Immunology Research Center, Medical school,Inflammation and Inflammatory Diseases Division,Mashhad University of Medical Sciences,Mashhad,Iran , , Amel Jamehdar، Saeid نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,Mashhad University of Medical Sciences,Mashhad,Iran , , Akbari Eydgahi، Mohammad Reza نويسنده Biotechnology Research Center,Semnan University of Medical Sciences,Semnan,Iran , , Sadeghian، Hamid نويسنده Department of Laboratory Sciences,Mashhad University of Medical Sciences,Mashhad,Iran , , Ghazvini، Kiarash نويسنده Antimicrobial Resistance Research Center, Bu-Ali Research Institute, School of Medicine,Department of Microbiology and Virology,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.
