Paper Information

Journal:   AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)   JANUARY-MARCH 2013 , Volume 5 , Number 1 (16); Page(s) 20 To 28.
 
Paper:  EXPRESSION, PURIFICATION AND CHARACTERIZATION OF THREE OVERLAPPING IMMUNODOMINANT RECOMBINANT FRAGMENTS FROM BORDETELLA PERTUSSISFILAMENTOUS HEMAGGLUTININ
 
Author(s):  ASGARIAN OMRAN HOSSEIN, AMIRZARGAR ALI AKBAR, ARJMAND HOSSEIN, ESHRAGHIAN MOHAMMADREZA, NIKBIN BEHROOZ, ESHRAGHI SAEID, MAHDAVI MARZIEH, KHOSHNOODI JALAL, JEDDI TEHRANI MAHMOOD, RABBANI HODJATALLAH, SHOKRI FAZEL*
 
* DEPARTMENT OF IMMUNOLOGY, SCHOOL OF PUBLIC HEALTH, TEHRAN UNIVERSITY OF MEDICAL SCIENCES, TEHRAN, IRAN
 
Abstract: 
Background: Filamentous hem agglutinin (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-
g production were assessed in a limited number of healthy adults vaccinated with a commercial a cellular 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-
g production.
Conclusion: Our preliminary results suggest that these overlapping recombinant FHA fragments are immunogenic and may prove to be immunoprotective.
 
Keyword(s): BORDETELLA PERTUSSIS, FILAMENTOUS HEMAGGLUTININ, IMMUNODOMINANT, PROKARYOTIC EXPRESSION, RECOMBINANT ANTIGEN
 
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