title: Expression of antigenic determinants of the haemagglutinin large subunit of novel influenza virus in insect cells.
abstract: The global outbreak of novel A/H1N1           spread in human population worldwide has revealed           an emergency need for producing a vaccine against           this virus. Current influenza vaccines encounter           problems with safety issues and weak response in           high-risk population. It has been established that           haemagglutinin is the most important viral antigen           to which antibody responses are directed, and recombinant           subunit vaccines, haemagglutinin of influenza           A and B viruses, have been considered in order           to facilitate vaccine production. In the present           study, we have focused on construction of a recombinant           baculovirus encoding the large subunit of           novel influenza virus A/H1N1 haemagglutinin. The           full genome of haemagglutinin was cloned into           pGEM-TEasy vector and sequenced. The large subunit           of the haemagglutinin gene was amplified by           PCR using specific primers and cloned into pFast-           BacHTc donor plasmid, which was then confirmed           by restriction enzyme analysis and sequencing and           transformed into E. coli DH10Bac competent cells.           The bacmid DNA was transfected into insect cells to           produce recombinant baculovirus. Expression of recombinant           haemagglutinin in insect cells was determined           by SDS-PAGE and immunoblotting. It has           been shown that the recombinant haemagglutinin           (rHA) obtained from the baculovirus insect cell expression           system has suitable immunogenicity in human           and can be considered as a candidate flu vac-           cine. Here we produced large amounts of the HA1           protein of novel influenza A/H1N1 (Iranian isolate)           in insect cells. The immunogenicity and efficacy of           the recombinant HA1 will be evaluated as a vaccine           candidate and compared to the recombinant HA1           produced in a prokaryotic system.
authors.affiliation: Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Science and Research Branch of Islamic Azad University, Tehran, Iran.
authors.forename: A
authors.initials: A
authors.lastname: Yousefi
authors.forename: F
authors.initials: F
authors.lastname: Fotouhi
authors.forename: S
authors.initials: S
authors.lastname: Hosseinzadeh
authors.forename: M T
authors.initials: MT
authors.lastname: Kheiri
authors.forename: B
authors.initials: B
authors.lastname: Farahmand
authors.forename: S
authors.initials: S
authors.lastname: Montazeri
authors.forename: F
authors.initials: F
authors.lastname: Mousavi
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D000939:Epitopes / Q000378:metabolism*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D018411:Spodoptera / Q000235:genetics / Q000821:virology; D014162:Transfection
chemical_list: D000939:Epitopes; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins