Determine the minimal level of replication required for broad protective immunity of influenza vaccine

项目摘要 甲型流感病毒导致5%-20%的人口患病，每年有20,000多人住院 我们。流感病毒由于快速进化而产生的抗原漂移和转移对每年一度的流感构成了严峻的挑战。 疫苗接种计划，这取决于对流感血清型的准确预测 将在下一个流感季节传播。最近流感疫苗的失败和潜在的暴发 流感大流行突显了对能够提供广泛保护的疫苗的迫切需要。干扰素 干扰素是天然免疫系统的重要组成部分，也是天然免疫系统和获得性免疫系统之间的桥梁。 免疫反应。我们最近用定量和定量的方法研究了流感基因组的抗干扰素功能。 高通量基因组学系统。通过在流感基因组中加入8个对干扰素敏感的突变，我们 产生了一种超干扰素敏感(HIS)病毒作为候选疫苗。他的病毒被高度减毒 野生型和免疫缺陷的SCID小鼠，但完全胜任IFNAR缺陷的小鼠。他提供了 针对同源和异源病毒挑战的保护。我们的中心假设是系统性的 消除病毒基因组多个片段上的干扰素逃避功能会产生适当的诱导 先天免疫反应，这是建立长期记忆B细胞反应和T细胞必不可少的 对流感减毒活疫苗的应答。我们的目标是确定最小复制 流感病毒减毒活疫苗所需的能力，识别和产生单轮 感染His病毒，在体外可以诱导强的IFNR信号，但在体内没有复制能力 活体由于先天免疫反应。这种疫苗候选病毒将限制一轮 在免疫期间感染，而干扰素诱导活性将强到足以非法 广泛的保护性免疫。我们将产生对干扰素高度敏感的病毒，这种病毒在 并鉴定其在肺上皮细胞中的复制动力学和对干扰素的反应性。在我们获得 这样的病毒，我们将用疫苗候选病毒感染小鼠，并表征诱导的免疫反应。 最后，我们将确定疫苗候选病毒对不同流感病毒株的保护效果。 体内的病毒。这一项目取得的成果将促进我们对流感疫苗的理解 开发和促进通用流感疫苗的开发。

Project Summary Influenza A virus causes disease in 5%-20% of the population with over 200,000 hospitalizations annually in US. The antigen drift and shift of influenza virus due to rapid evolution pose a serious challenge for annual flu vaccination program, which is effective depending on the accurate prediction of the influenza serotypes that will be circulating in the next flu season. The recent failure of influenza vaccine and potential outbreak of influenza pandemics highlights the urgent need for a vaccine that can provide broad protection. Interferon (IFN) is a critical component of the innate immune system and also the bridge between the innate and adaptive immune responses. We recently studied the anti-IFN function of influenza genome using a quantitative and high-throughput genomics system. By incorporating eight IFN-sensitive mutations into influenza genome, we generated a Hyper Interferon Sensitive (HIS) virus as a vaccine candidate. HIS virus is highly attenuated in wild type and immune-deficient SCID mice, but fully competent in IFNAR-deficient mice. HIS provides protection against homologous and heterologous viral challenges. Our central hypothesis is that systematical elimination of IFN-evasion functions on multiple segments of the virus genome generates proper induction of innate immune response, which is essential for establishing long term memory B cell response and T cell response by live attenuated influenza vaccine. Our objective is to determine the minimal replication capacity required for live attenuated influenza virus vaccine, identify and generate single-round infection HIS virus, which can induce strong IFNR signaling in vitro, but has no replication capacity in vivo due to innate immune response. Such vaccine virus candidate would have confined one-round infection during immunization whereas the IFN inducing activity would be strong enough to illicit broad protective immunity. We will generate hyper IFN sensitive virus that has no replication capacity in vivo, and characterize its replication kinetics and responsiveness to IFN in lung epithelial cells. After we obtain such virus, we will infect mice with vaccine candidate viruses and characterize the induced immune responses. Finally, we will determine protection efficacy of vaccine candidate viruses against different strains of influenza virus in vivo. The results achieved from this project will advance our understanding of influenza vaccine development and facilitate the development of universal influenza vaccine.