Oral vaccine

口服疫苗
  • 文章类型: Journal Article
    背景:登革热是一种快速传播的蚊媒热带病毒性疾病,每年影响全球数亿人。登革热病毒(DENV)包括四种遗传上不同的血清型,它们会导致严重的危及生命的感染,包括登革热出血热/登革热休克综合征。登革热疫苗的开发由于预先存在的交叉反应性抗体依赖性增强而导致疫苗增强的严重登革热疾病的可能性而变得复杂。以及同型抗体。因此,尽管进行了多年的研究,但尚未开发出有效的登革热疫苗,该疫苗可对四种DENV血清型中的每一种同时和持久的免疫。在受资源限制严重的国家进行大规模免疫接种,口服疫苗被认为比常规方法更有益。因此,在设计经济有效的候选疫苗的持续努力中,本研究应用酵母表面展示技术,利用全重组酵母细胞开发口服登革热疫苗候选物,该酵母细胞展示了与合成共有登革热包膜结构域III(scEDIII)融合的M细胞靶向配体Co1的重组融合蛋白。用重组酵母细胞口服饲喂雌性Balb/c小鼠,并监测系统和粘膜免疫反应的免疫原性。
    结果:使用登革热特异性抗体和异硫氰酸荧光素缀合的抗小鼠IgG抗体的免疫荧光显微镜清楚地表明,与没有荧光的scEDIII-Co1和Mock细胞相比,重组蛋白Co1-scEDIII-AGA位于相应克隆的细胞表面。表面显示的Co1-scEDIII-AGA的口服剂量应用以登革热特异性血清IgG的形式刺激了全身性体液免疫反应,以及分泌性免疫球蛋白A(sIgA)形式的粘膜免疫反应。来自脾脏和Peyer斑块的分离淋巴样细胞中的抗原特异性B细胞反应进一步支持了升高的粘膜免疫反应。此外,与使用纯化的scEDIII抗原腹膜内加强后的scEDIII-Co1和Mock相比,表面显示的Co1-scEDIII-AGA喂养引起强烈的免疫反应。
    结论:与未展示的scEDIII-Co1相比,表面展示的Co1-scEDIII-AGA制剂诱导强免疫原性。先前的研究已经支持scEDIII构建体对所有四种血清型的中和潜力。因此,口服展示生物活性Co1-scEDIII融合蛋白的基因工程酵母全细胞,无需任何进一步加工,显示出作为抗登革病毒感染的有效口服候选疫苗的前景.
    BACKGROUND: Dengue is a rapidly spreading mosquito borne tropical viral disease affecting hundreds of millions of people across the globe annually. The dengue virus (DENV) includes four genetically distinct serotypes that cause serious life-threatening infections, including dengue hemorrhagic fever/dengue shock syndrome. Dengue vaccine development is complicated by the possibility of vaccine-enhanced severe dengue disease due to antibody-dependent enhancement by pre-existing cross-reactivity, as well as homotypic antibodies. Thus, the development of an efficacious dengue vaccine conferring simultaneous and durable immunity to each of the four DENV serotypes has not yet been developed despite years of research. For mass immunization in deeply affected resource-limited countries, oral vaccination is considered more beneficial than conventional approaches. Therefore, in a continuing effort towards designing economical and potent vaccine candidates, the current study applied yeast surface display technology to develop an oral dengue vaccine candidate using whole recombinant yeast cells displaying the recombinant fusion protein of M cell targeting ligand Co1 fused to the synthetic consensus dengue envelope domain III (scEDIII). Female Balb/c mice were orally fed with recombinant yeast cells and immunogenicity in terms of systemic and mucosal immune responses was monitored.
    RESULTS: Immunofluorescence microscopy with dengue specific antibody and fluorescein isothiocyanate-conjugated anti-mouse IgG antibody clearly showed that recombinant protein Co1-scEDIII-AGA was localized on the cell surface of the respective clones in comparison with scEDIII-Co1 and Mock cells with no fluorescence. Oral dosage applications of surface displayed Co1-scEDIII-AGA stimulated a systemic humoral immune response in the form of dengue-specific serum IgG, as well as a mucosal immune response in the form of secretory immunoglobulin A (sIgA). Antigen-specific B cell responses in isolated lymphoid cells from the spleen and Peyer\'s patches further supported an elevated mucosal immune response. In addition, surface displayed Co1-scEDIII-AGA feeding elicited strong immune responses in comparison with scEDIII-Co1 and Mock following intraperitoneal booster with purified scEDIII antigen.
    CONCLUSIONS: Surface displayed preparations of Co1-scEDIII-AGA induced strong immunogenicity compared with non-displayed scEDIII-Co1. Prior studies have supported the neutralization potential of scEDIII constructs against all four serotypes. Thus, the oral administration of genetically engineered yeast whole cells displaying biologically active Co1-scEDIII fusion protein without any further processing shows prospective as a potent oral vaccine candidate against dengue viral infection.
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  • 文章类型: Journal Article
    Dengue virus (DENV) infection is an emerging global health threat. DENV consists of four distinct serotypes, necessitating a tetravalent vaccine. In this study, expression of consensus envelope protein domain III (cEDIII) fused to cholera toxin B subunit (CTB) in transgenic rice calli was improved using the luminal binding protein BiP at the N-terminus and the SEKDEL signal sequences at the C-terminus, targeting the recombinant protein to endoplasmic reticulum (ER). We found that the fusion protein showed higher levels of expression when compared to the fusion proteins using rice amylase 3D (RAmy3D) or CTB native signal sequence only. The CTB-cEDIII fusion protein was evaluated as an oral dengue vaccine candidate in mice. Serotype specific systemic IgG antibodies and specific IgA response in feces were detected and furthermore, T cell proliferation and high frequency antibody-secreting B cells were detected in the spleen. These results suggest the possible use of plant-based dengue tetravalent vaccine targeted to the mucosal immune system for induction of systemic and mucosal immune responses to DENV infection.
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