Approach to control caries through apple-derived vaccine

描述：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果为基础的疫苗，可用于人类对龋齿（变形链球菌）的免疫。我们将通过开发转基因苹果（Malus domestica）生产可食用的疫苗试剂来实现这一目标。具体目的是获得植物双表达载体，携带由S. mutans Agl/ll与霍乱毒素（CT） A2和B亚基（CTA2B）偶联的唾液结合区（SBR）和/或葡萄糖基转移酶（GTF）葡聚糖结合（GLU）蛋白组成的嵌合蛋白进行农杆菌介导转化，获得表达SBR-CTA2B和/或GLU基因的转基因苹果株系。我们将设计用于农杆菌介导植物转化的植物二元表达载体，将SBR-CTA2B和/或GLU基因导入苹果基因组。待验证的假设是，SBR-CTA2B和/或GLU在转基因苹果中表达，苹果中表达的SBR-CTA2B和GLU蛋白协同引起小鼠的保护性免疫反应。首先，我们将获得转基因苹果植株，产生具有抗原性的SBR-CTA2B和GLU蛋白，并定量在体外转基因苹果中产生SBR-CTA2B和/或GLU蛋白的量。转基因苹果叶片产生SBR-CTA2B和/或GLU蛋白后，小鼠免疫产生保护性免疫反应。我们将测试转基因苹果叶组织中表达的SBR-CTA2B和/或GLU蛋白是否诱导小鼠血清和粪便中的IgG和IgA以及唾液中的IgA反应。我们还将研究SBR-CTA2/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、有用的以水果为基础的预防龋齿疫苗的输送系统。这项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递送系统，用于治疗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.