title: CFP10: mFcγ2 as a novel tuberculosis vaccine candidate increases immune response in mouse.
abstract: OBJECTIVE
  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.
  
  
  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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Ali Asghar
authors.initials: AA
authors.lastname: Baghani
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Saman
authors.initials: S
authors.lastname: Soleimanpour
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Hadi
authors.initials: H
authors.lastname: Farsiani
authors.affiliation: HIV/AIDS, HTLV and Viral Hepatitis Research Center, Iranian Academic Center for Education, Culture and Research (ACECR), Mashhad, Iran.
authors.forename: Arman
authors.initials: A
authors.lastname: Mosavat
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Yousefi
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Meshkat
authors.affiliation: Immunology Research Center, Inflammation and Inflammatory Diseases Division, Medical school, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Seyed Abdolrahim
authors.initials: SA
authors.lastname: Rezaee
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Saeid Amel
authors.initials: SA
authors.lastname: Jamehdar
authors.affiliation: Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran.
authors.forename: Mohammad Reza Akbari
authors.initials: MR
authors.lastname: Eydgahi
authors.affiliation: Organic Chemistry, Department of Laboratory Sciences, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Sadeghian
authors.affiliation: Antimicrobial Resistance Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Kiarash
authors.initials: K
authors.lastname: Ghazvini
keywords: CFP-10; Fcγ2a; Immunogenicity; Mycobacterium – tuberculosis; Subunit vaccine