title: [Construction and immunogenicity of recombinant bacteriophage T7 vaccine expressing M2e peptides of avian influenza virus].
abstract: To construct a recombinant T7 phage expressing matrix protein 2 ectodomain (M2e) peptides of avian influenza A virus and test immunological and protective efficacy in the immunized SPF chickens. M2e gene sequence was obtained from Genbank and two copies of M2e gene were artificially synthesised, the M2e gene was then cloned into the T7 select 415-1b phage in the multiple cloning sites to construct the recombinant phage T7-M2e. The positive recombinant phage was identified by PCR and sequencing, and the expression of surface fusion protein was confirmed by SDS-PAGE and Western-blot. SPF chickens were subcutaneously injected with 1 X 10(10) pfu phage T7-M2e, sera samples were collected pre- and post-vaccination, and were tested for anti-M2e antibody by ELISA. The binding capacity of serum to virus was also examined by indirect immunofluorescence assay in virus- infected CEF. The immunized chickens were challenged with 200 EID50 of H9 type avian influenza virus and viral isolation rate was calculated to evaluate the immune protective efficacy. A recombinant T7 phage was obtained displaying M2e peptides of avian influenza A virus, and the fusion protein had favorable immunoreactivity. All chickens developed a certain amount of anti-M2e antibody which could specially bind to the viral particles. In addition, the protection efficacy of phage T7-M2e vaccine against H9 type avian influenza viruses was 4/5 (80%). These results indicate that the recombinant T7 phage displaying M2e peptides of avian influenza A virus has a great potential to be developed into a novel vaccine for the prevention of avian influenza infection.
authors.affiliation: National Veterinary Biological Medicine Engineering Research Center, Nanjing 210014, China. hai-x@126.com
authors.forename: Hai
authors.initials: H
authors.lastname: Xu
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Wang
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Tang
authors.forename: Qi-Sheng
authors.initials: QS
authors.lastname: Zheng
authors.forename: Ji-Bo
authors.initials: JB
authors.lastname: Hou
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D017123:Bacteriophage T7 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D015967:Gene Expression Regulation, Viral; D007114:Immunization; D009980:Influenza A virus / Q000235:genetics / Q000276:immunology*; D007252:Influenza Vaccines / Q000276:immunology*; D005585:Influenza in Birds / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D010455:Peptides / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins; D013047:Specific Pathogen-Free Organisms; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000086082:Viroporin Proteins
chemical_list: D000914:Antibodies, Viral; D007252:Influenza Vaccines; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins