Production and characterization of recombinant pertactin, fimbriae 2 and fimbriae 3 from Bordetella pertussis.
BACKGROUND
Bordetella pertussis is a causative agent of pertussis or whooping cough in humans. Pertactin (Prn), fimbriae 2 (Fim2) and fimbriae 3 (Fim3) of B. pertussis are important virulence factors and immunogens which have been included in some acellular pertussis vaccines. In this present study, we cloned, expressed and purified Prn, Fim2 and Fim3, respectively. The immunogenicity and protective efficacy of the three recombinant proteins (rPrn, rFim2 and rFim3) were investigated in mouse model.


RESULTS
Three recombinant proteins with amount of 12 to 25 mg/L were produced. Compared to the control mice only immunized with adjuvant, serum IgG antibody responses were significantly induced in the mice immunized with rPrn, rFim2 or rFim3 (P < 0.001 for all three proteins). Furthermore, T cell responses characteristic of increased production of IL-2 and TNF-alpha (only for rPrn) were elicited in the mice immunized with the three proteins (P < 0.05 for all three proteins). Immunization with rPrn, but not with rFim2 or rFim3, significantly enhanced clearance of bacteria in the lungs of mice after intranasal challenge with B. pertussis (P < 0.05). When tested in a lethal intracerebral infection model, certain protection was observed in mice immunized with rPrn.


CONCLUSIONS
We have developed an efficient method to produce large amounts of rPrn, rFim2, and rFim3 from B. pertussis. The three recombinant proteins induced both humoral and cellular immune responses in mice. Immunization with rPrn also conferred protection against pertussis in mouse infection models. Our results indicated that the recombinant proteins still retain their immunological properties and highlighted the potential of the recombinant proteins for the future development of the B. pertussis vaccines.
Xu
Yinghua
Y
Department of serum, National Institute for the Control of Pharmaceutical and Biological Products, Temple of Heaven, Beijing 100050, PR China. jxyhxyh@yahoo.com.cn
Wang
Yaying
Y
Tan
Yajun
Y
Zhang
Huajie
H
Wu
Lijie
L
Wang
Lichan
L
Hou
Qiming
Q
Zhang
Shumin
S
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000276:immunology*; D001886:Bordetella pertussis / Q000276:immunology* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D036781:Fimbriae Proteins / Q000096:biosynthesis / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D007376:Interleukin-2 / Q000097:blood; D008297:Male; D051379:Mice; D010567:Pertussis Vaccine / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000097:blood; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis / Q000276:immunology*; D014917:Whooping Cough / Q000276:immunology / Q000517:prevention & control
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007074:Immunoglobulin G; D007376:Interleukin-2; D010567:Pertussis Vaccine; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D010566:Virulence Factors, Bordetella; D036781:Fimbriae Proteins; C068929:pertactin