title: [Characterization the immunogenicity of recombinant VP2 of infectious bursal disease virus containing N-terminal M2e of avian influenza virus].
abstract: OBJECTIVE
  We developed subunit vaccines against H5 or H9 subtype avian influenza viruses (AIV) and infectious bursal disease viruses (IBDV). Viral protein 2 (VP2) of IBDV was used as cargo protein to display a 12-amino-acid (aa) immunodominant epitope derived from N-terminal M2 extracelluar domain (nM2e) of H5 or H9 subtype AIV.
  
  
  METHODS
  The aa and nucleotide sequence of nM2e was determined by comparing the available avian influenza vaccine strains and alignment the AIV sequence available in GenBank. One copy of H5 or H9 nM2e was inserted into P(BC) region of VP2 origin from IBDV B87 vaccine strain by fusion polymerase chain reaction. The VP2(BC)nM2e recombinants were cloned into Bac-to-Bac expression system and transfected to Sf9 cell. The expressed chimeric protein was characterized by indirect immunofluorescence assay and Western blotting, and subsequently was used as antigen to develop vaccine. The non-immunized chicken was given two injections with the vaccine at a 4-week interval. Serum against VP2 and nM2e was tested by indirect ELISA and virus neutralization in chick embryo fibroblast.
  
  
  RESULTS
  Both VP2(BC)nM2e recombinants were successfully constructed and expressed in Sf9 cell. Both chimeric proteins elicited antibody against VP2 and nM2e. The antibody level elicited by VP2(BC)nM2e(H5) vaccine was higher than that of VP2(BC)nM2e(H9).
  
  
  CONCLUSIONS
  Both chimeric proteins were immunigenic, and the efficacy of VP2(BC)nM2e(H5) was higher than VP2(BC)nM2e(H9) chicken.
authors.affiliation: Jiangsu Academy of Agricultural Sciences, National Research Center of Engineering and Technology for Veterinary Bioproducts, Nanjing 210014, China. tyhlw@yahoo.com.cn
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Tang
authors.forename: Yuzhen
authors.initials: Y
authors.lastname: Gong
authors.forename: Yongwei
authors.initials: Y
authors.lastname: Wang
authors.forename: Jihu
authors.initials: J
authors.lastname: Lu
authors.forename: Peipei
authors.initials: P
authors.lastname: Wu
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Jiahui
authors.initials: J
authors.lastname: He
authors.forename: Jibo
authors.initials: J
authors.lastname: Hou
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000276:immunology; D018175:Birnaviridae Infections / Q000821:virology; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D005347:Fibroblasts / Q000276:immunology; D007243:Infectious bursal disease virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007252:Influenza Vaccines / Q000276:immunology / Q000494:pharmacology; D005585:Influenza in Birds / Q000276:immunology / Q000473:pathology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014765:Viral Vaccines / Q000276:immunology* / Q000494:pharmacology
chemical_list: D007252:Influenza Vaccines; D011993:Recombinant Fusion Proteins; C082513:VP2 protein, infectious bursal disease virus; D014764:Viral Proteins; D015678:Viral Structural Proteins; D014765:Viral Vaccines