Novel vita-vaccine formula combines safety of dead and efficacy of live vaccines

建议书摘要 减毒活疫苗已被证明是对许多严重传染性疾病最有效的人类疫苗。 疾病。与死亡疫苗相比，活疫苗产生了更好的免疫保护和 永恒的记忆。但是，尽管活疫苗有效，但对其安全性的担忧导致了疫苗拒绝接种 由于一些人和拒绝对非常年轻的人、老年人和免疫系统进行管理。 很难保存和运送活疫苗，特别是向发展中国家的贫困地区 而且很贵。了解活疫苗疗效的分子基础具有重要意义，因为它 将能够靶向相关的免疫途径，从而产生最佳和持久的保护 豁免权。重要的是，它将为开发安全且负担得起的疫苗奠定基础。 作为活疫苗进行保护，缓解公众的恐惧，并增加人口的比例，即 接种了疫苗。八年前，我们从先天免疫细胞感知微生物的假设开始我们的研究。 作为一套独特的病原体相关分子模式(PAMP)，我们鉴定了细菌 信使RNA(MRNA)作为VitA-PAMP，标志着细菌的生存能力和动员定制的免疫 对于死亡的微生物，不需要做出回应。Toll样受体(TLR)信号适配器TRIF发挥着 在炎症I型干扰素和NLRP3炎症体通路的上游起中心作用。添加 死亡细菌的细菌mRNA重述了这些先天反应，并在死亡疫苗中添加了 细菌信使核糖核酸(我们称之为维生素疫苗)增强了它在小鼠身上的表现。一种维他命疫苗的研制 与活疫苗类似，在独特地诱导滤泡T辅助细胞反应(帮助B细胞)、生发 中心形成和B细胞同型类转换，都是以TRIF依赖的方式进行的。这些研究提供了 强有力的证据表明，现有疫苗的VitA疫苗版本可能代表着 能够将活疫苗的效力与死亡疫苗的安全性结合起来。我们的三个重叠区域 将在此项目中调查的项目包括： 1.我们将确定如何通过补充死亡细菌疫苗来引发适应性免疫 VITA-PAMP细菌mRNA与PAMP如细菌脂肽等PAMP所诱导的mRNA进行比较。 2.我们将研究细菌mRNA如何影响亚单位疫苗的性能。我们将测试VITA- 获得许可的炭疽亚单位疫苗和甲型流感病毒单价亚单位疫苗的疫苗版本。 3.我们将测试三价灭活流感病毒疫苗的VitA疫苗版本，并将其与 流感减毒活疫苗。 这些研究的完成应提供充分的实验证据，以保证使用 细菌mRNAs作为优良的维生素佐剂，恢复死亡疫苗的微生物活性特征 并改进现有的灭活疫苗和亚单位疫苗，以防止细菌或病毒疾病。

PROPOSAL SUMMARY Live attenuated vaccines have proven to be the most efficient human vaccines for many serious infectious diseases. When compared to their dead counterparts, live vaccines induce superior immune protection and lasting memory. But despite the efficacy of live vaccines, concerns over their safety have led to vaccine refusal by some and withholding their administration to the very young, the elderly and immunocompromised. Preservation and delivery of live vaccines especially to impoverished areas in developing countries is difficult and expensive. Understanding the molecular basis for the efficacy of live vaccines is significant because it would enable targeting of the relevant immune pathways that induce optimal and long-lasting protective immunity. Importantly, it would set the stage for the development of vaccines that are safe and afford the same protection as live vaccines, alleviating public fears and increasing the segment of the population that is vaccinated. We began our work eight years ago with the hypothesis that innate immune cells sense microbial viability as a distinct set of pathogen associated molecular patterns (PAMPs), and we identified bacterial messenger RNA (mRNA) as a vita-PAMP that signifies bacterial viability and mobilizes a tailored immune response not warranted for dead microorganisms. The Toll-like receptor (TLR) signaling adaptor TRIF plays a central role here upstream of inflammatory type I interferon and NLRP3 inflammasome pathways. Adding bacterial mRNA to dead bacteria recapitulates these innate responses, and supplementing a dead vaccine with bacterial mRNA (what we call a vita-vaccine) augments its performance in mice. A vita-vaccine performed similarly to a live vaccine in uniquely eliciting a follicular T helper cell response (that helps B cells), germinal center formation, and B cell isotype class switching, all in a TRIF-dependent manner. These studies provide strong evidence that vita-vaccine versions of existing vaccines could represent a significant advance in being able to combine the efficacy of live vaccines with the safety of dead vaccines. The three overlapping areas we will investigate in this project are: 1. We will determine how adaptive immunity elicited by the supplementation of a dead bacterial vaccine with the vita-PAMP bacterial mRNA compares to that elicited by PAMPs such as bacterial lipopeptides and others. 2. We will investigate how bacterial mRNA impacts the performance of subunit vaccines. We will test vita- vaccine versions of the licensed anthrax subunit vaccine and Influenza A virus monovalent subunit vaccine. 3. We will test a vita-vaccine version of a trivalent inactivated Influenza virus vaccine and compare it to the live attenuated influenza vaccine. The completion of these studies should provide sufficient experimental evidence to warrant the use of bacterial mRNAs as superior vita-adjuvants that restore the signatures of microbial viability to dead vaccines and improve existing inactivated and subunit vaccines for protection against either bacterial or viral diseases.