Intrinsically-enhanced Ebola and Marburg virus like particles for increased potency and immune memory

摘要 这一开发项目寻求开发方法来显著提高病毒样颗粒的免疫原性。 (VLP)疫苗。埃博拉病毒(EBOV)和马尔堡病毒(Marv)是衣壳类丝状病毒家族的成员， 引起严重人类疾病爆发的负义RNA病毒。由于它们的致命性，丝状病毒 是为数不多的需要生物安全级别4(BSL4)控制的新兴病毒之一 学习。尽管在EBOV疫苗方面取得了进展，但仍有必要开发 其他丝状病毒的疫苗，特别是MARV。VLP是一种经常探索的疫苗平台，适用于各种 病毒，包括EBOV和MARV。VLP的优势包括相对安全，因为它们不能复制 并导致病毒性疾病；它们与正宗病毒非常相似，以至于它们在自己的母语中呈现抗原 状态；它们诱发B和T细胞反应的能力，以及它们在各种系统中被证明的有效性。 与活病毒相比，VLP作为疫苗的潜在挑战包括免疫原性相对较低。 丝状病毒VLP可由病毒基质蛋白VP40、病毒糖蛋白(GP)、 有或不表达病毒核蛋白(NP)。这种VLP已被证明能刺激 树突状细胞(DC)反应，诱导B和T细胞介导的免疫并保护小鼠、豚鼠和非 人类灵长类动物(NHP)免受致命挑战。因此，丝状病毒VLP是可行的候选疫苗。然而， 丝状病毒VLP的免疫原性较弱，需要与佐剂和多次联合给药才能保护 剂量。这对于主要为应对疫情而部署的疫苗来说并不理想。此应用程序寻求 通过合理掺入I型来显著提高EBOV和MARV VLP的免疫原性 干扰素(干扰素)信号域从细胞模式识别受体RIG-I或TRIF进入颗粒。 这使得VLP能够将干扰素诱导区域的细胞传递到细胞中，从而触发强大的先天免疫 使人联想到活病毒感染的反应，以增强适应性免疫反应。为了进一步追求这一点 方向，我们将探索优化干扰素诱导信号域整合到EBOV的方法 和马夫VLP。然后，我们将评估不同增强型VLP策略的能力，以获得B和T 体内的细胞免疫反应，最后测试最有希望的增强型VLP引发快速 对EBOV和MARV攻击小鼠的保护作用。这些研究的成功完成将提供一个 开发丝状病毒疫苗的新平台和一种可能应用于其他 新出现的病毒病原体。

Summary This developmental project seeks to develop methods to dramatically boost immunogenicity of virus-like particle (VLP) vaccines. Ebola virus (EBOV) and Marburg virus (MARV) are members of the filovirus family of enveloped, negative-sense RNA viruses that causes outbreaks of severe human disease. Due to their lethality, filoviruses are among the handful of emerging viruses for which biosafety level 4 (BSL4) containment is required for their study. Although progress has been made toward EBOV vaccines, there remains a need for development of vaccines for other filoviruses, particularly MARV. VLPs are an often-explored vaccine platform for a variety of viruses, including for EBOV and MARV. Advantages of VLPs include their relative safety, as they cannot replicate and cause viral disease; their close resemblance to authentic virus, such that they present antigens in their native state; their capacity to elicit both B and T cell responses and their proven efficacy for a variety of systems. Potential challenges of VLPs as vaccines include relatively lower immunogenicity, as compared to live virus. Filovirus VLPs can be produced by co-expression of the viral matrix protein VP40, the viral glycoprotein (GP), with or without expression of the viral nucleoprotein (NP). Such VLPs have been demonstrated to stimulate dendritic cell (DC) responses, to elicit B and T cell mediated immunity and to protect mice, guinea pigs and non- human primates (NHPs) from lethal challenge. Therefore, filovirus VLPs are viable vaccine candidates. However, filovirus VLP immunogenicity is weak with protection requiring coadministration with an adjuvant and multiple doses. This is not ideal for vaccines that are mainly deployed in response to outbreaks. This application seeks to dramatically improve immunogenicity of EBOV and MARV VLPs by the rational incorporation of type I interferon (IFN) signaling domains from the cellular pattern recognition receptors RIG-I or TRIF into the particles. This enables delivery into cells of the IFN inducing domain by the VLPs, triggering robust innate immune responses reminiscent of live virus infection, to boost adaptive immune responses. To further pursue this direction, we will explore approaches to optimize the incorporation of IFN-inducing signaling domains into EBOV and MARV VLPs. We will then evaluate the capacity of the different enhanced VLP strategies to elicit B and T cell immune responses in vivo, and finally, test the most promising enhanced VLPs for the capacity to elicit rapid protection against EBOV and MARV challenge in mice. Successful completion of these studies will provide a novel platform for the development of filovirus vaccines and an approach that can likely be applied to other emerging viral pathogens.