Mechanism of membrane inactivation method to prepare enveloped virus vaccines

项目摘要/摘要 世界卫生组织列出的构成最高公共健康风险的优先病原体名单，以及 没有足够的治疗方法，完全是由包膜病毒组成的。高度包膜病毒的例子 与人类健康有关的有：艾滋病毒、甲型流感病毒、EBOV、MERS-CoV、SARS、SARS-CoV-2、CHIKV、 新城疫病毒、戊型肝炎病毒、呼吸道合胞病毒、流行性出血热病毒以及甲型和乙型流感病毒。他们的疾病造成的人类代价是 数百万美元，每年的经济成本高达数千亿美元。的标准方法 产生疫苗，包括全灭活病毒(WIV)、减毒活病毒(LAV)、亚单位和 DNA/RNA，往往不能导致有效的预防。WIV疫苗有一些优点，如安全性 以及它们将多种抗原呈递给免疫系统。然而，WIV经常导致不足 由于WIV疫苗之间的抗原构象差异而导致的免疫力甚至疫苗增强型疾病 和挑战病毒。我们的建议满足了对更有效和更广泛适用的迫切需求 通过探索一种使用抗病毒药物XM-01的有前景的新技术来开发WiV疫苗(专利申请正在申请中)。 目前用于WIV制备的病毒灭活方法使用化学或物理方法，通常会损坏或 修饰病毒抗原，包括糖蛋白，这是包膜病毒的重要免疫原。 这通常会导致非保护性甚至破坏性的免疫反应。例如， 目前用于IAV的WIV疫苗在10%-60%之间。我们最近开发了一种新的疫苗平台 使用XM-01靶向病毒膜，同时在很大程度上保持病毒的天然构象 糖蛋白。以IAV为模型的初步数据表明，XM-01灭活IAV免疫 颗粒提高了对血凝素(HA)和神经氨酸酶(NA)的中和抗体的诱导，AS 与传统的福尔马林灭活的IAV颗粒相比，动物存活率更高。由于XM-01是一种广泛的- 包膜病毒的光谱抑制剂，我们的中心假设是XM-01可以改善病毒灭活 通过保留糖蛋白构象，促进有效地产生 保护性免疫反应。我们将通过两个具体目标来验证我们的假设： 目的1：确定XM-01灭活病毒增强免疫反应的机制 与传统的WIV疫苗方法相比，使用的是流感病毒模型。 目的2：确定XM-01疫苗接种的安全性、有效性和免疫应答的广度。 我们期待这项工作将加深我们对XM-01膜介导的机制的理解。 病毒灭活会产生增强的保护性免疫反应，为了评估新方法的安全性， 针对异源病毒株的有效性和产生广泛保护性抗体的能力，全面评估 这种新的灭活模式是否适用于WIV疫苗的广泛改进。

Project Summary / Abstract The World Health Organization’s list of priority pathogens that pose the highest public health risks, and for which there are insufficient treatments, is composed solely of enveloped viruses. Examples of enveloped viruses highly relevant to human health are: HIV, influenza A viruses (IAV), EboV, MERS-CoV, SARS, SARS-CoV-2, CHIKV, NiV, HeV, RSV, ZIKV, and the influenza A (IAV) and B (IBV) viruses. The human cost of their illnesses is in the millions, and the economic cost is in the hundreds of billions of dollars per year. The standard methods for generating vaccines, including whole inactivated virus (WIV), live attenuated virus (LAV), sub-unit, and DNA/RNA, often fail to result in effective prophylaxis. WIV vaccines have some advantages such as their safety and their presentation of multiple antigens to the immune system. However, WIVs often induce insufficient immunity or even vaccine-enhanced disease due to antigen conformational differences between the WIV vaccine and the challenge virus. Our proposal addresses the pressing need for more effective and broadly applicable WIV vaccines by exploring a promising new technology using the antiviral XM-01 (patent applications pending). Current virus inactivation methods for WIV preparation use chemical or physical means that often damage or modify viral antigens, including the glycoproteins, which are important immunogens for the enveloped viruses. This often leads to non-protective or even destructive immune responses. For example, the efficacies of the currently used WIV vaccines for IAV are between 10%-60%. We recently developed a novel vaccine platform that uses XM-01 to target the viral membrane while largely preserving the native conformation of viral glycoproteins. Preliminary data using IAV as a model indicates that immunization with XM-01-inactivated IAV particles improved induction of neutralizing antibodies to both hemagglutinin (HA) and neuraminidase (NA), as well as animal survival, compared to the traditional formalin-inactivated IAV particles. As XM-01 is a broad- spectrum inhibitor of enveloped viruses, our central hypothesis is that virus inactivation with XM-01 improves WIV vaccine development by preserving glycoprotein conformations, facilitating the effective generation of a protective immune response. We will test our hypothesis with two Specific Aims: Aim 1: Determine the mechanism by which XM-01 inactivated virus affords improved immune responses as compared to traditional WIV vaccine methods, using the influenza virus model. Aim 2: Determine XM-01 vaccination safety, efficacy, and breadth of immune responses. We expect this work will improve our understanding of the mechanism by which XM-01 membrane-mediated viral inactivation generates an enhanced protective immune response, and to assess the new method’s safety, efficacy, and ability to generate broadly protective antibodies against heterologous viral strains, overall assessing whether this new mode of inactivation may be amenable for broad improvement of WIV vaccines.