Mechanism of membrane inactivation method to prepare enveloped virus vaccines

项目总结/摘要 世界卫生组织的优先病原体清单，构成最高的公共卫生风险， 目前还没有足够的治疗方法，它只由包膜病毒组成。包膜病毒高度 与人类健康有关的是：艾滋病毒，甲型流感病毒（IAV），EboV，MERS-CoV，SARS，SARS-CoV-2，CHIKV， NiV、HeV、RSV、ZIKV以及甲型流感（IAV）和B（IBV）病毒。他们的疾病的人类成本是在 数百万，每年的经济成本高达数千亿美元。标准方法 产生疫苗，包括完整灭活病毒（WIV）、活减毒病毒（LAV）、亚单位，以及 DNA/RNA，往往不能导致有效的预防。WIV疫苗具有一些优点，例如其安全性 以及它们向免疫系统呈递多种抗原。然而，WIV往往会导致不足 由于WIV疫苗之间的抗原构象差异， 和挑战病毒我们的建议针对迫切需要更有效和更广泛适用的 通过探索一种使用抗病毒XM-01的有前途的新技术（专利申请正在进行中）来接种WIV疫苗。 目前用于WIV制备的病毒灭活方法使用化学或物理手段，其经常损害或破坏WIV的生物活性。 修饰病毒抗原，包括糖蛋白，其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， 目前使用的用于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 它使用XM-01靶向病毒膜，同时在很大程度上保留了病毒的天然构象， 糖蛋白使用IAV作为模型的初步数据表明，用XM-01灭活的IAV免疫 颗粒改善了对血凝素（HA）和神经氨酸酶（NA）的中和抗体的诱导， 以及动物的存活率，与传统的福尔马林灭活的IAV颗粒相比。由于XM-01是一个广泛的- 作为包膜病毒的广谱抑制剂，我们的中心假设是XM-01的病毒灭活提高了 通过保留糖蛋白构象，促进有效产生 保护性免疫反应我们将用两个具体目标来检验我们的假设： 目的1：确定XM-01灭活病毒提供改善免疫应答的机制 与传统的WIV疫苗方法相比，使用流感病毒模型。 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。 我们希望这项工作将提高我们对XM-01膜介导的细胞凋亡机制的理解。 病毒灭活产生增强的保护性免疫应答，为了评估新方法的安全性， 有效性和产生针对异源病毒株的广泛保护性抗体的能力， 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.