A Novel Mucosal Vaccine Delivery System Using Bacteriophage T4 Nanoparticles

 描述(由申请人提供)：在这个项目中，我们寻求填补黏膜疫苗交付方面的一个重要空白。大多数病原体通过黏膜表面进入人体，如鼻腔和阴道通道以及肠道，但到目前为止，几乎所有疫苗都是注射到肌肉或皮下。这些疫苗刺激全身免疫反应，但往往会使黏膜上皮--我们抵御感染的主要屏障--变得脆弱不堪。我们建议开发一种使用T4噬菌体衣壳的新型且适应性强的黏膜疫苗平台。这些衣壳可以同时包装疫苗DNA，呈现蛋白质抗原，并在头部表面显示靶向配体。因此，我们可以设计一种有针对性的疫苗载体，为免疫系统提供“蛋白质启动”和“DNA增强”，以产生持久的免疫反应。为此，我们将使用细胞表面受体Toll样受体2和5(TLR2&5)将T4-Head靶向上皮细胞。这些受体是早期 先天免疫反应，是疫苗佐剂的靶标。触发先天免疫反应是发展适应性、持久反应的关键。我们还将优化编码疫苗DNA的转基因盒子，以便在上皮细胞中高水平和长期表达。我们假设，通过将头部定位于粘膜上皮并将它们输送到粘膜相关组织，我们将在粘膜上皮细胞中引发系统而显著的强烈免疫反应。为了验证这一假设，我们将靶向-T4衣壳递呈抗原和携带疫苗-DNA到小鼠舌下(舌下)、鼻腔和肌肉内。然后我们将比较早期(第3天)和晚期(1年)的先天免疫反应和获得性免疫反应。我们希望在早期先天反应最强的群体中观察到最强大的适应性免疫反应。我们预测，所有给药途径都会产生全身反应，但我们将看到舌下和鼻腔给药组的黏膜免疫反应最强。如果成功，这项工作将产生一个有效的、新颖的、粘膜疫苗平台，生产成本低，并易于适应多种疾病。

 DESCRIPTION (provided by applicant): In this project, we seek to fill an important gap in mucosal vaccine delivery. Most pathogens enter the body at mucosal surfaces such as the nasal and vaginal passageways and the intestines, yet nearly all vaccines to date are injected into the muscle or under the skin. These vaccines stimulate a systemic immune response, but often leave the mucosal epithelium, our primary barrier to infection, vulnerable. We propose to develop a novel and adaptable mucosal vaccine platform using T4-bacteriophage capsids. These capsids can simultaneously package vaccine DNA, present protein antigens, and display targeting ligands on the head surface. Therefore, we can engineer a targeted vaccine vector that presents the immune system with a "protein-prime" and a "DNA-boost" to generate a long-lasting immune response. To do this, we will target the T4-heads to epithelial cells using the cell-surface receptors toll-like receptor 2 and 5 (TLR2 & 5). These receptors are part of the early innate immune response and are targets of vaccine adjuvants. Triggering an innate immune response is key to developing an adaptive, long-lasting response. We will also optimize a transgene cassette encoding vaccine DNA for high-level and long-term expression in epithelial cells. We hypothesize that by targeting the heads to the epithelium and delivering them to mucosal associated tissues, we will elicit a strong immune response systemically and significantly, in the mucosal epithelium. To test this hypothesis, we will deliver targeted-T4 capsids presenting antigen and carrying vaccine-DNA to mice sublingually (under-the-tongue), intranasally, and intramuscularly. We will then compare the innate and adaptive immune responses at both early (day 3) and late time-points (1 year). We expect to observe the most robust adaptive immune response in the groups with the strongest early innate response. We predict that all routes of administration will generate a systemic response, but we will see the most robust mucosal immune response in the sublingual and the intranasal groups. If successful, the work here will generate an effective, novel, mucosal vaccine platform that is inexpensive to produce and readily adaptable to a multitude of diseases.