Construction of recombinant BCG expressing the vp1 antigen of Enterovirus 71 for the development of a candidate vaccine

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dc.contributor.authorNurulhasanah, Othman
dc.date.accessioned2014-11-11T08:30:01Z
dc.date.available2014-11-11T08:30:01Z
dc.date.issued2007
dc.descriptionMasteren_US
dc.description.abstractEnterovirus 71 (EV71), the causativeing agent of outbreaks of hand, foot and mouth disease (HFMD) in children is identified as a current outbreak that needsrequire urgent control due to the higher high number of cases. Vaccination is one of the most effective methods to control the disease outbreaks. In this study, a recombinant BCG vaccine candidate was constructed against EV71. The recombinant BCG (rBCGV1) which expressesed a synthetic gene encoding theed VP1 protein of EV71 fused to ubiquitin complex (UbGR) which was constructed amplified using the technique of assembly PCR. The synthetic gene was codon technique after optimized for expressioning the genes into mycobacterium codon bias. The Ag85A promoter and signal peptide peptide sequence from M.tuberculosis drove was used to drive the expression and secretion of the synthetic gene expression was from M.tuberculosis. Hence, Tthe expression of the VP1-UbGRUbGR-VP1 fusion protein was revealed confirmed by Western blotting using rabbit polyclonal antibody specific to the VP1 protein and was found in the cell recombinant pellet cell by the used ofof the recombinant BCG rabbit polyclonal antibody which specific to VP1 protein. Mice BALB/c (H-2d) that was immunized with rBCGV1 cloned showed the significant the ability to induce moderate antibody production in serum in BALB/c (H-2d) mice when sera from immunized mice were , tested against purified VP1-UbGRUbGR-VP1 purified fusion protein. IgG2a subclass antibody was shown to be induced at a has a significantly greater higher level than IgG1. Spleenocytes, which was obtained from rBCGV1 , immunized mice showed significant higher level of lymphocyte proliferation when it was stimulated with VP1-UbGRUbGR-VP1 antigencompared to control. The Analyses of intracellular cytokines analysis show that, which was expressed by CD4+ T cells and CD8+ T cells from rBCGV1 immunized mice, showed the were stimulated by VP1-UbGR antigen UbGR-VP1 protein to stimulated CD4+T cells to express significant leves of IL-2, IFN-γ and IL-4 when compared to the control. This antigen also stimulated CD8+T cell to express IL-2 and IFN-γ. EThe extracellular cytokine analyseis also showed the significantly higher levels of IFN-γ when it was stimulated with VP1- UbGR antigencompared to control. Overall, the immunogenicity studies results suggested that the rBCGV1 enhanced the stimulation of immune system towards the Th1 (T helper 1) pathway against the studied antigen. Data from this study also suggested the potential of rBCGV1 to be developed as a vaccine and further studies must be carriedy on out to evaluate the efficacy of this candidate vaccine.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/428
dc.language.isoenen_US
dc.subjectBiological Scienceen_US
dc.subjectBCGen_US
dc.titleConstruction of recombinant BCG expressing the vp1 antigen of Enterovirus 71 for the development of a candidate vaccineen_US
dc.typeThesisen_US
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