Enhancement of immune responses against Iranian isolate of FMD-type O/IRN/1/2010 based on VP1 and human HSP70 genes and comparison with conventional vaccine
Sedeh, Farahnaz MotamediYazdanpanah, SolmazSoleimanjahi, HooriehMahravani, HoomayonShafaee, KamalodinRazavi, Mohammad HadiRezaee, Abass
Background: Foot and mouth disease (FMD) is the causative agent and one of the most transmissible livestock diseaseswhich cause important economic losses. The genome of FMD virus is a positive-sense, single-stranded RNA and it wrappedwith 60 copies of 4 structural proteins, VP1, VP2, VP3 and VP4. The VP1 is a major immunogenic antigen with criticalepitopes for inducing immune responses. The current vaccine, which successfully prevents disease, includes inactivatedwhole virus antigen. However, it is not without problems. The aim of this study is enhancement of immune responsesagainst Iranian isolate of FMD-type O/IRN/1/2010 based on VP1 and human HSP70 fusion protein in BALB/c mice.Materials, Methods & Results: In this study FMD virus type O/IRN/2010 was isolated from vesicles of the infected cattlein Qoum, and serotyped as a new linage of type O (PanAsia-linage 2) in Iran. The isolated FMD virus was propagated onIBRS2 cell line and whole RNA of the infected cells was extracted by commercial kit instruction. The extracted RNA wasamplifi ed using VP1 gene-specifi c primer pairs by means of one-step RT-PCR. The specifi c primer pair was designed byAllelID6 software. There are sequences of Kpn I and BamH I restriction enzymes and three overhanging nucleotides atthe start of forward and reverse primers, respectively. The VP1 nucleotide sequence was deposit in Genbank-NCBI database under accession number JN 676146. The purifi ed VP1 gene was sub-cloned into PTZ57R/T vector. Then digestedVP1 gene by KpnI and BamHI enzymes was ligated in pcDNA3.1+ vector as a DNA vaccine. Also, the improved DNAimmunization system was constructed using pcDNA3.1+ plasmid contains VP1 gene of Iranian isolate FMD virus typeO/IRN/1/2010 and human HSP70 gene and expression of VP1-HSP70 fusion protein confi rmed in BHKT7 cell line byGuinea pig specifi c polyclonal antibody against FMD virus type O and conjugated rabbit...(AU)
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