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

项目摘要 本R01提案的目标包括疫苗佐剂的分子和免疫学特征。 与干扰素基因蛋白刺激物(STING)介导的过程相关的活性。疫苗 由惰性抗原组成，通常只有弱的免疫原性，因此需要联合注射佐剂 以增强免疫刺激作用。不幸的是，很少有佐剂被批准用于临床，而且 佐剂相关增强的机制还知之甚少。刺是一种模式识别 检测由细菌环化酶或细菌环化酶合成的含嘌呤环二核苷酸(CDN)的受体 宿主酶环状GMP-AMP(CGAMP)合成酶(CGAS)在其感应胞浆DNA之后。 抗体和细胞介导的STING药理激活可显著增强疫苗效力 对不同微生物病原体的保护。此外，内源性合成的cGAMP是天然的 从外体和病毒颗粒中的细胞中释放出来。我们通过构建细胞系来利用这种现象 我们用来产生免疫原性病毒样颗粒(VLP)。 我们现在证明了来自无关的新出现的黄病毒和甲病毒的VLP有效地包裹了生物活性 CDN和刺激体内的刺痛依赖活性。由于细胞外CDN高度易感 从注射部位迅速降解和分散，我们假设这种简单的方法代表 在排水中利用刺痛导向佐剂效应的变革性技术改进 淋巴结节。我们还预测这将是一个高度适应性的平台，并建议探索 它适用于可能出现的病毒类型。在实验上，这也代表了一种独特的调查 该模型将允许在体内表征获得性免疫的先天和分子相关性，这些相关性是 通过有针对性的刺痛活动来加强。因此我们假设，抗原导向免疫的增强 基于刺激剂的佐剂的反应在功能上与关键的分子和细胞过程有关，这些过程是 使用我们建议的疫苗和实验系统可以识别。我们计划使用集成的 使用分子转录分析、免疫图谱的小鼠模型和非人类的方法 我们团队设计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.