A Self-Adjuvanting Virus Like Particle Vaccine Platform for Emerging Viruses

项目摘要 该R 01提案的目标包括疫苗佐剂的分子和免疫学表征 与干扰素基因刺激因子（STING）蛋白介导的过程相关的活性。疫苗 包含惰性抗原通常仅具有弱免疫原性，因此需要佐剂的共同施用 以增强免疫刺激。不幸的是，很少有佐剂被批准用于临床使用， 对促排剂相关增强的机制知之甚少。STING是一种模式识别 一种检测由细菌环化酶或 宿主酶环GMP-AMP（cGAMP）合酶（cGAS）在其感测胞质DNA之后。 STING的药理学活化极大地增强了疫苗效力，如抗体和细胞介导的免疫反应所示。 保护免受各种微生物病原体的侵害。此外，内源性合成的cGAMP是天然的。 从细胞外泌体和病毒颗粒中释放出来。我们利用这一现象， 其表达组成型活性环化酶，然后我们用其产生免疫原性病毒样颗粒（VLP）。 我们现在表明，来自不相关的新兴黄病毒和甲病毒的VLP有效地包裹了生物活性物质， CDN和刺激体内STING依赖性活性。由于细胞外CDN非常容易受到 降解并迅速从注射部位分散，我们假设这种简单的方法代表了 一种变革性的技术改进，用于在引流中利用STING导向的辅助作用， 淋巴结我们还预测这将是一个高度适应性的平台，并建议探索在多大程度上 它适用于可能出现的病毒类型。实验上，这也代表了一个独特的研究 该模型将允许在体内表征适应性免疫的先天和分子相关性， 通过靶向STING活性增强。因此，我们假设，抗原导向免疫的增强 基于STING的佐剂的免疫应答在功能上与关键的分子和细胞过程有关， 使用我们提出的疫苗和实验系统。我们计划使用一个集成的 一种使用分子转录组学分析的方法，免疫分析的小鼠模型，以及非人类的 病毒生长和保护性免疫的灵长类动物模型。

Project Summary The goals of this R01 proposal include the molecular and immunological characterization of vaccine adjuvant activity associated with processes mediated by the protein Stimulator of Interferon Genes (STING). Vaccines comprised of inert antigen are often only weakly immunogenic and thus require co-administration of adjuvants to augment immunostimulation. Unfortunately, very few adjuvants are approved for clinical use and the mechanisms of adjuvant-associated enhancement are poorly understood. STING is a pattern recognition receptor that detects purine-containing cyclic dinucleotides (CDN) synthesized by either bacterial cyclases or the host enzyme cyclic GMP-AMP (cGAMP) synthase (cGAS) following its sensing of cytosolic DNA. Pharmacologic activation of STING greatly enhances vaccine efficacy as indicated by antibody and cell-mediated protection against diverse microbial pathogens. Furthermore, endogenously synthesized cGAMP is naturally released from cells in exosomes and virus particles. We exploited this phenomenon by constructing cell lines that express constitutively active cyclases that we then used to produce immunogenic virus-like particles (VLP). We now show that VLP derived from unrelated emerging flavi- and alphaviruses efficiently encapsulate bioactive CDN and stimulate STING-dependent activity in vivo. Since extracellular CDNs are highly susceptible to degradation and disperse quickly from the site of injection, we hypothesize that this simple approach represents a transformative technological improvement for harnessing STING-directed adjuvant effects in the draining lymph node. We also predict this will be a highly adaptable platform and propose to explore the extent to which it is applicable to potentially emerging virus types. Experimentally, this also represents a unique investigative model that will allow in vivo characterization of the innate and molecular correlates of adaptive immunity that are potentiated by targeted STING activity. We thus hypothesize that the enhancement of antigen-directed immune responses by STING-based adjuvants is functionally linked to key molecular and cellular processes that are discernable using our proposed vaccines and experimental systems. We plan to explore this using an integrated approach that uses molecular transcriptomic analysis, mouse models of immune profiling, and nonhuman primate models of viral growth and protective immunity devised by our group.