title: Expression, Purification and Characterization of Three Overlapping Immunodominant Recombinant Fragments from Bordetella pertussis Filamentous Hemagglutinin.
abstract: BACKGROUND
  Filamentous hemagglutinin (FHA) is one of the most important immunoprotective antigens of Bordetella pertussis (B. pertussis) and a major component of the acellular pertussis vaccine. In the present study, three overlapping recombinant fragments from the immunodominant region of FHA were produced and their immunogenicity was investigated.
  
  
  METHODS
  Three overlapping coding sequences of FHA antigen were amplified from B. pertussis genomic DNA by PCR. Amplified fragments were expressed in Escherichia coli (E. coli) BL21(DE3) strain and purified through His-tag using Nickel-based chromatography. Purified fragments were characterized by SDS-PAGE and Western blotting techniques. In vitro peripheral blood mononuclear cells (PBMC) proliferation and IFN-γ production were assessed in a limited number of healthy adults vaccinated with a commercial acellular pertussis vaccine in response to all purified FHA fragments by H3-Thymidine incorporation and ELISA, respectively.
  
  
  RESULTS
  Recombinant FHA segments were successfully cloned and produced at high levels in E. coli BL21(DE3). SDS-PAGE and Western blot analyses confirmed their purity and reactivity. All three recombinant fragments together with a commercial native FHA were able to induce in vitro PBMC proliferation and IFN-γ production.
  
  
  CONCLUSIONS
  Our preliminary results suggest that these overlapping recombinant FHA fragments are immunogenic and may prove to be immuno-protective.
authors.affiliation: Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran ; Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Asgarian-Omran
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Amirzargar
authors.forename: Mohammad
authors.initials: M
authors.lastname: Arjmand
authors.forename: Mohammadreza
authors.initials: M
authors.lastname: Eshraghian
authors.forename: Behrooz
authors.initials: B
authors.lastname: Nikbin
authors.forename: Saeid
authors.initials: S
authors.lastname: Eshraghi
authors.forename: Marzieh
authors.initials: M
authors.lastname: Mahdavi
authors.forename: Jalal
authors.initials: J
authors.lastname: Khoshnoodi
authors.forename: Mahmood
authors.initials: M
authors.lastname: Jeddi-Tehrani
authors.forename: Hodjatallah
authors.initials: H
authors.lastname: Rabbani
authors.forename: Fazel
authors.initials: F
authors.lastname: Shokri
keywords: Bordetella pertussis; Filamentous hemagglutinin; Immunodominant; Prokaryotic expression; Recombinant antigen