Author/Authors :
Asgarian Omran، Hossein نويسنده , , Amirzargar، Ali Akbar نويسنده , , Arjmand، Mohammad نويسنده Department of Biochemistry, Pasteur Institute of Iran , , Eshraghian ، Mohammadreza نويسنده Department of Biostatistics and Epidemiology, School of Public Health, Tehran University of Medical Sciences , , Nikbin، Behrooz نويسنده , , Eshraghi، Saeid نويسنده Department of Microbiology, School of Public Health, Tehran University of Medical Sciences , , Mahdavi، Marzieh نويسنده Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Khoshnoodi، Jalal نويسنده Department of Immunology, School of Public Health, Tehran University of Medical Sciences , , Jeddi Tehrani]، Mahmood نويسنده , , Rabbani، Hodjattallah نويسنده Immune and Gene Therapy Laboratory, CCK, Karolinska University Hospital Solna Department of Antigen and Antibody Engineering, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Shokri، Fazel نويسنده ,
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.
Conclusion: Our preliminary results suggest that these overlapping recombinant FHA fragments are immunogenic and may prove to be immuno-protective.
