title: [Immunogenicity and protective efficacy of pertactin recombinants against Bordetella bronchiseptica challenge].
abstract: OBJECTIVE
  In this study we showed the immunogenicity and protective efficacy of five pertactin recombinants against Bordetella bronchiseptica (Bb) challenge.
  
  
  RESULTS
  The complete coding sequence (2040 bp) of the prn gene (PRN) and its fragments,5'-terminal 1173 bp fragment (PN),3'-terminal 867 bp fragment (PC), two copies of region I (654 bp; PR I) in PN, and 2 copies of region II (678 bp; PR II) in PC, were separately cloned into the prokaryotic expression vector pGEX-KG, and expressed in the Eschierichia coli BL21 (DE3) using induction by isopropyl-beta-D-thiogalactopyranoside. The recombinant proteins were named GST-PRN, GST-PN, GST-PC, GST-2PR I and GST-2PR II. All five recombinant proteins showed immunological reactivity in the Western-blot analysis. Mice, immunized subcutaneously with two doses of the purified proteins mixed with an equal volume of Freund's adjuvant,produced robust PRN-specific IgG antibody levels. When challenged, 6 of 9 mice in GST-2PR I group and all 9 mice in the other groups survived intranasal challenge with three times the 50% lethal dose (LD50) of virulent Bb HH0809. After challenge with 10 LD50 7/9,3/9,6/9,1/10 and 6/10 of the mice survived. Furthermore, complete protection against intraperitoneal (i.p.) challenge with 10 LD50 of HH0809 was observed in mice that were injected i.p. with 0.5 ml rabbit anti-GST-PRN, GST-PN,GST-PC or GST-2PR II serum. Only 1 of 10 mice survived in the group of mice that received anti-GST-2PR I, and no survivors were noted in the group of mice that received PRN-absorbed rabbit antiserum (0/5).
  
  
  CONCLUSIONS
  In this study,we showed that all of five pertactin recombinants had differential immunogenicity and protective efficacy against Bb challenge. Mice immunized with GST-PC had better survival against fatal Bb challenge than did those immunized with GST-PN. In addition, GST-2PR II and GST-2PR I provided the similar results These data may have implications for the development of safe and efficacious subunit vaccines for the prevention of bordetellosis on the basis of these five pertactin recombinants.
authors.affiliation: Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Techology, China. zhaozhanqin@126.com
authors.forename: Zhanqin
authors.initials: Z
authors.lastname: Zhao
authors.forename: Chen
authors.initials: C
authors.lastname: Wang
authors.forename: Yun
authors.initials: Y
authors.lastname: Xue
authors.forename: Ke
authors.initials: K
authors.lastname: Ding
authors.forename: Chunjie
authors.initials: C
authors.lastname: Zhang
authors.forename: Xiangchao
authors.initials: X
authors.lastname: Cheng
authors.forename: Yinju
authors.initials: Y
authors.lastname: Li
authors.forename: Yichen
authors.initials: Y
authors.lastname: Liu
authors.forename: Tingcai
authors.initials: T
authors.lastname: Wu
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D016950:Bordetella bronchiseptica / Q000276:immunology*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; C068929:pertactin