Correlates of protective immunity to HCV and rational vaccine design: Project 3

丙型肝炎病毒(丙型肝炎病毒)继续感染着全球约7100万人，估计新增了150万至200万人 每年都有感染。彻底根除丙型肝炎病毒感染需要开发一种有效的疫苗， 能迅速诱导持久的抗病毒免疫，预防感染。来自自发控制者的证据 提示广泛和持久的丙型肝炎病毒特异性的CD4和CD8T细胞应答对有效控制至关重要 丙型肝炎病毒感染。此外，对病毒糖蛋白E1和E2的抗体具有中和保护作用。 也有牵连。此外，在肝脏(病毒的主要部位)产生抗病毒免疫是至关重要的 复制)。因此，这项提议的主要目标是开发一种使用DNA修饰的疫苗接种策略 安卡拉牛痘(MVA)和基于蛋白质的疫苗，将产生强大和广泛的丙型肝炎病毒特异性T和B 血液和肝脏对多种丙型肝炎病毒蛋白的细胞反应。我们假设一个强大的，广泛的， 和血液和肝脏中针对丙型肝炎病毒抗原的持续T细胞反应结合强烈的诱导 中和抗体反应将提供对丙型肝炎病毒感染和不同组合的保护 可以利用DNA/MVA/蛋白质疫苗来实现所需的保护性免疫。我们建议 使用3个具体目标来实现这些目标。在目标1中，我们将构建和表征DNA和MVA 表达丙型肝炎病毒核心、NS3-NS5和E1E2蛋白的疫苗。在目标2中，我们将优化DNA/MVA疫苗 诱导强T细胞和抗体反应的方式。优化将包括在 病毒样颗粒(VLP)上或分泌出两种形式。此外，我们还将测试 CD40L佐剂对细胞免疫和体液免疫的影响。在目标3中，我们将针对DNA/MVA优化E1E2蛋白启动子 进一步增强抗体反应的疫苗。通过完成这些研究，我们希望产生一个 最理想的抗丙型肝炎病毒疫苗，能诱导强大的T细胞和中和抗体反应。

Hepatitis C virus (HCV) continues to infect ~71 million people worldwide with an estimated 1.5 to 2 million new infections each year. A complete eradication of HCV infection warrants development of an effective vaccine that can rapidly induce a durable anti-viral immunity and prevent infection. Evidence from spontaneous controllers suggests that a broad and durable HCV-specific CD4 and CD8 T cell responses are critical for effective control of HCV infection. In addition, a protective role for neutralizing antibody against viral glycoproteins E1&E2 has also been implicated. Furthermore, it is critical generate anti-viral immunity in the liver (the primary site of virus replication). Thus, the primary goal of this proposal is to develop a vaccination strategy using DNA, modified vaccinia Ankara (MVA), and protein-based vaccines that will generate a robust and broad HCV specific T and B cell responses in blood and liver against multiple HCV proteins. We hypothesize that induction of a strong, broad, and persistent T cell response against HCV antigens in the blood and liver combined with induction of strong neutralizing antibody response will provide protection against HCV infection and different combinations of DNA/MVA/protein vaccination can be harnessed to achieve the desired protective immunity. We propose to achieve these objectives using 3 specific aims. In Aim 1 we will construct and characterize DNA and MVA vaccines expressing HCV core, NS3-NS5 and E1E2 proteins. In Aim 2 we will optimize the DNA/MVA vaccine modality for inducing strong T cell and antibody responses. The optimizations will include testing the E1E2 in two forms either presented on the virus-like particle (VLP) or secreted. In addition, we will test the influence of CD40L adjuvant on cellular and humoral immunity. In Aim 3 we will optimize E1E2 protein boosts for DNA/MVA vaccine for further enhancing the antibody responses. By completion of these studies we hope generate an optimal vaccine against HCV that induces strong T cell and neutralizing antibody responses.