Approach to control caries through apple-derived vaccine

产品说明：我们的研究项目的目标是生产安全，廉价和易于管理的苹果水果为基础的疫苗，可用于人类免疫预防龋齿，（变形链球菌）。我们将通过开发生产可食用疫苗试剂的转基因苹果来实现这一目标。本研究的具体目的是获得植物双元表达载体，以携带由沙门氏菌唾液结合区组成的嵌合蛋白。将突变体Ag 1/11与霍乱毒素（CT）A2和B亚基（CTA 2 B）和/或葡糖基转移酶（GTF）的葡聚糖结合（GLU）蛋白偶联，用于农杆菌介导的转化，以获得表达SBR-CTA 2 B和/或GLU基因的转基因苹果系。我们将设计植物双元表达载体，用于农杆菌介导的植物转化，将SBR-CTA 2B和/或GLU基因导入苹果基因组。待测试的假设是SBR-CTA 2B和/或GLU在转基因苹果中表达，并且在苹果中表达的SBR-CTA 2B和GLU蛋白在小鼠中协同增强保护性免疫应答。首先，我们将获得转基因苹果植物以产生抗原性SBR-CTA 2B和GLU蛋白，并定量体外转基因苹果中产生的SBR-CTA 2B和/或GLU蛋白的量。在用转基因苹果叶产生SBR-CTA 2B和/或GLU蛋白免疫的小鼠中将观察到小鼠中的保护性免疫应答。我们将测试转基因苹果叶组织中表达的SBR-CTA 2B和/或GLU蛋白是否诱导小鼠的血清和粪便IgG和伊加以及唾液伊加应答。我们还将研究SBR-CTA 2/B和GLU口服免疫在保护小鼠龋齿方面是否存在协同作用。长期目标是开发一种潜在的和有用的基于水果的递送系统用于龋齿疫苗。拟议的研究将开创一种新的方法，开发一种基于植物果实的疫苗输送系统，用于预防龋齿和其他传染病的药物疫苗。

DESCRIPTION: The goal of our research project is to produce safe, inexpensive and easily administered apple fruit-based vaccine which can be used in humans for immunization against Dental caries, (Streptococcus mutatis). We will accomplish this by developing transgenic apple (Malus domestica) producing the edible vaccine reagent. Specific Aims are to obtain plant binary expression vectors to carry chimeric protein consisting of the saliva-binding region (SBR) of S. mutans Agl/ll coupled to cholera toxin (CT) A2 and B subunits (CTA2B) and/or glucan-binding (GLU) protein of glucosyltransferases (GTF) for Agrobacterium-mediated transformation, to obtain transgenic apple line to express SBR-CTA2B and/or GLU genes. We will design the plant binary expression vector for Agrobacterium-mediated plant transformation to deliver SBR-CTA2B and/or GLU genes to apple genome. Hypotheses to be tested are that the SBR-CTA2B and/or GLU are expressed in transgenic apples and both SBR-CTA2B and GLU proteins expressed in apple synergistically elicits protective immune responses in mice. First, we will obtain transgenic apple plant to produce antigenic SBR-CTA2B and GLU proteins and quantify the amount of SBR-CTA2B and/or GLU proteins produced in in vitro transgenic apples. Protective immune response in mice will be observed in mice immunized by transgenic apple leaf producing SBR-CTA2B and/or GLU proteins. We will test if the SBR-CTA2B and/or GLU proteins expressed in transgenic apple leaf tissue induces serum and fecal IgG and IgA, and salivary IgA responses in mice. We also will investigate whether there is synergy between SBR-CTA2/B and GLU oral immunization with respect to protection to Dental caries in mice. The long range goal is to develop a potential and useful fruit-based delivery system for a vaccine against Dental caries. The proposed research will initiate a novel approach to develop a plant fruit-based vaccine delivery system for a pharmaceutical vaccine against Dental caries and other infectious diseases.