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

丙型肝炎病毒（HCV）继续感染全世界约7100万人，估计有150万至200万新感染者。 每年感染。彻底根除HCV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力，预防感染。来自自发控制者的证据 表明广泛而持久的HCV特异性CD 4和CD 8 T细胞应答对于有效控制HCV感染至关重要。 HCV感染。此外，中和抗体对病毒糖蛋白E1和E2的保护作用， 也受到牵连。此外，在肝脏（病毒的主要部位）中产生抗病毒免疫是至关重要的 复制）。因此，该提议的主要目标是开发使用修饰的DNA的疫苗接种策略， 安卡拉牛痘（MVA）和基于蛋白质的疫苗，其将产生稳健和广泛的HCV特异性T和B 血液和肝脏中的细胞对多种HCV蛋白的反应。我们假设，诱导一个强大的，广泛的， 血液和肝脏中针对HCV抗原的持续T细胞应答与诱导强的 中和抗体应答将提供针对HCV感染的保护， DNA/MVA/蛋白质疫苗接种可用于实现所需的保护性免疫。我们建议 通过三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HCV核心、NS 3-NS 5和E1 E2蛋白的疫苗。目标2：优化DNA/MVA疫苗 用于诱导强T细胞和抗体应答的模式。优化将包括测试E1 E2， 存在于病毒样颗粒（VLP）上或分泌的两种形式。此外，我们还将测试 CD 40 L佐剂对细胞和体液免疫的影响。在目标3中，我们将优化DNA/MVA的E1 E2蛋白增强 用于进一步增强抗体应答的疫苗。通过完成这些研究，我们希望产生一个 最佳的抗HCV疫苗，诱导强烈的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.